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The Remarkable Journey of Medical Genetics Pioneer, Arno Motulsky

Dr. Arno G. Motulsky, recognized for having pioneered the field of “pharmacogenetics”—the genetic basis for differences in the way individuals respond to drugs—passed away on January 17, 2018 in Seattle, at the age of 94. Since Motulsky was not a virologist, his passing is noted here not for his scientific contributions but, rather, for his improbable achievements against very long odds and, especially, for the remarkable journey from which this story began—a journey which brings to mind a current issue before the public. It began as follows.

Dr. Arno Motulsky in his Seattle apartment in 2014. Credit Clare McLean/University of Washington

1938 saw a dramatic increase in violent anti-Jewish acts by the Nazi regime in Germany. In May 1939, 916 passengers, including 910 German Jews seeking to escape from the Nazis, were aboard the SS St. Louis; a liner of the Hamburg-America Line, which was bound for Cuba. Hundreds of thousands of other German Jews would not be able to get out.

Each of the passengers on the St. Louis had a visa from the Cuban government that was issued before the ship departed from Germany. However, unbeknownst to the passengers—but known to the shipping line—the President of Cuba, Federico Laredo, had invalidated the visas the day before the ship sailed. Thus, when the St. Louis arrived in Havana Harbor, the Cuban government would not allow the passengers to disembark. Instead, the Cuban authorities attempted to exploit the refugees’ situation in order to extort bribes from them. These bribes began at $500 a person and rose to $1,000,000! All the while the ship sat within yards of the shore.

Cuba had been practicing this sort of extortion before the incident involving the St. Louis. Manuel Benitez Gonzalez, the Director-General of the Cuban immigration office, routinely sold landing documents for $150 or more and, according to U.S. estimates, had amassed a personal fortune of $500,000 to $1,000,000. Eventually, Benitez’s corruption would lead to his resignation. However, corruption was not the only factor working against the refugees. Many Cubans saw them as competitors for scarce jobs. Moreover, Cuban sympathy for the refugees’ plight was also tempered by xenophobia and antisemitism. [News reports of the impending sailing of the St. Louis triggered anti-Semitic demonstrations in Cuba even before the ship left Germany.]

As the St. Louis sat at anchor in Havana Harbor, the American public raptly followed daily media accounts of the refugees’ plight. And despite urgent appeals from American sympathizers (non-Jewish as well as Jewish), the U.S. State Department and the White House decided not to intervene on behalf of the refugees. President Franklin Roosevelt did not answer telegrams from the refugees on the ship. The State Department did respond, wiring back to the passengers that they must “await their turns on the waiting list and qualify for and obtain immigration visas before they may be admissible into the United States.”

Roosevelt had the power to issue an executive order that would have allowed the St. Louis’ refugees to enter the United States. He had, in fact, permitted 15,000 German and Austrian Jews, who were in the United States on visitor’s permits, to stay, saying: “It would be a cruel and inhuman thing to compel them to leave here” for Germany.   

Why then did Roosevelt and the State Department abandon the refugees on the St. Louis? American popular opinion, which was opposed to accepting more new arrivals, was a key factor. Indeed, about 80% of Americans were against easing immigration restrictions. A key reason was that the depression left many Americans out-of-work. Consequently, many were fearful of foreigners becoming competitors for scarce American jobs. Antisemitism, isolationism (in the years after World War I, many Americans were staunchly against the U.S. being drawn into European affairs), and xenophobia were no doubt factors as well.

The National Origins Act of 1924 permitted only a fraction of would-be emigres from Germany to enter the United States. The quotas that were set on countries were intended to favor “desirable” immigrants from Northern and Western Europe, while limiting less “racially desirable” immigrants, such as Southern Europeans, Eastern European Jews, and people born in Asia and Africa. Yet anti-German sentiment may have led American immigration officials to allow only one fourth of the German quota to be filled. Also note that three months before the St. Louis sailed to Cuba, the U.S. Congress let die a bill that would have admitted 20,000 Jewish children from Germany above the existing quota.

Critics contend that despite the political pressure on Roosevelt, he might have done more to raise public sympathy for the plight of the refugees. Moreover, once the United States was in the war, and the U.S. government became aware of Hitler’s intent to annihilate all Jews in Germany and in German controlled areas, the government tried to withhold that information from the public. And as the facts about the murder of the European Jews—concentration and extermination camps—leaked out, and despite pleas from Jewish leaders in Warsaw and elsewhere in Europe, Roosevelt would not act to impede the extermination program (e.g., by ordering the bombing of Auschwitz and the rail lines to it). Instead, Roosevelt maintained that winning the war was the only way to save the refugees, and winning the war was the absolute priority.

Barriers to Jewish immigrants were also in place across Latin America. [Note that the British refused to consider Palestine as a solution to the Jewish refugee problem.] Thus, Columbia, Chile, Paraguay, and Argentina rejected appeals from the Joint Distribution Committee (JDC, the major Jewish agency for relief overseas) for haven. And, when the JDC could not meet a deadline set by Cuba for the payment of ransoms, the St. Louis was forced to leave Havana Harbor. Remarkably, the German captain of the St. Louis, Gustav Schröder (who had been unaware of the Cuban government’s duplicity before the St. Louis sailed), appealed to the United States for haven. But his personal pleas were unheeded. What’s more, as the St. Louis sailed past Miami and the Florida coast, U.S. Coast Guard ships made certain that none of its passengers might escape to freedom. So, the St. Louis turned back to Europe, with sixteen-year-old Arno Motulsky among its passengers.

The German liner St. Louis being denied entrance to Havana, Cuba, in June 1939. Dr. Motulsky was aboard the ship with his mother and siblings when it was sent back to Europe. Credit Associated Press

But the St. Louis did not return to Germany. The JDC had arranged with Great Britain, France, Belgium, and the Netherlands for each of these countries to take one fourth of the refugees. Still, only the 288 refugees admitted to Great Britain would be safe, since the Nazis would overrun the other countries within months. Only a few would survive the Holocaust.

Motulsky was among the refugees who disembarked from the St. Louis in Belgium. He then spent a year in internment camps in Vichy France, where many other detainees died from starvation or typhoid. He reached the United States in June of 1941; ten days before his 18th birthday. His account of his flight to freedom, in his own words, is as follows:

“…By early 1939, my parents realized we would need to leave Germany. My dad had a brother in Chicago, so we hoped to go there, but in order to get out of Germany and into America, one needed a visa, which required a quota number that took a while to come up. So my father went to Cuba, where immigration permits were available, with the idea that we would join him later in Chicago. But over the next few months, as the situation deteriorated in Germany, my parents decided it would be best for my mother and brother and sister and me to join my father in Cuba and go to Chicago later.

We left Hamburg in May 1939, on a ship named the St. Louis. There were almost a thousand Jewish refugees on board. The voyage from Hamburg to Havana was normal. But before we could disembark in Havana, the Cubans declared our permits void and refused to let us land. The ship was German, but the captain, Gustav Schröder, was very sympathetic to us and sailed all over the Caribbean in search of a friendly port. Of course, we asked to land in America, but were denied. When we sailed close to Miami, US Coast Guard cutters and planes shooed us off.

So ultimately the ship had to return to Europe. I was 16, so still a little naive, but many of the passengers had been interned previously and knew what awaited us on our return. Some attempted suicide by jumping overboard. Telegrams were sent all over the world asking that we be allowed to disembark anyplace other than Germany. Miraculously, a few days before we would have arrived back in Germany, four other countries—England, France, Holland, and Belgium—each agreed to take one-fourth of the passengers. By luck of the draw, my mother and brother and sister and I were assigned to Belgium. So in June 1939, I started high school in Brussels.

Eleven months later, on May 1, 1940, my dad got his visa to move from Cuba to the US and went to Chicago. On the same day, our US visas came through as well. But by May 10, we hadn’t yet gotten out of Belgium, and the Germans invaded. Leaving was impossible. Since I was now 17, I was arrested by the Belgians as an enemy alien—ironically, as a German—and sent to an internment camp in France. As the Germans invaded France, we prisoners were moved further and further south, until we ultimately were interned in a camp in the Pyrenees, in Vichy-controlled France. The Vichy French, who collaborated with the Germans, sent the ethnic Germans back to Germany and kept the Jews interned. The internment camps had no food or sanitation. Many prisoners died, most from typhoid or starvation.

About ten months later, those of us with US visas were allowed to go to another camp, near Marseilles, where there was an American consulate. My visa had expired, but I pleaded for its renewal. It was ultimately granted, and in June 1941, ten days before my 18th birthday, I left France legally, crossed Spain into Portugal, and sailed from Lisbon to America. Ten days later and I wouldn’t have made it, because Franco did not allow males 18 or older to pass through Spain. A few months later, the Vichy French turned over all their internment camps to the Gestapo.

In August 1941, I arrived in America and joined my father in Chicago. Two years later, we learned that my mother and brother and sister had also survived. With the help of Belgian friends, they had crossed illegally into neutral Switzerland, where they were allowed to live unharmed until the end of the war. My family reunited in Chicago in early 1946.”

In Chicago, in 1942, 18-year-old Motulsky was examined on material he learned in the informal classes taught in the internment camps in France, and was then granted a high school certificate. Motulsky then enrolled in evening classes at Central YMCA College (which later became Roosevelt University) in Chicago, while working at a job during the day. He met his future wife, Gretel, in his English class at the college. He notes that Gretel was the better English student because she survived the war in England and America after leaving Germany in 1938. They were married in 1945.

Remarkably, in 1943, Motulsky was accepted by the medical school of the University of Illinois in Chicago. He also became a U.S. citizen, joined the Army, and was assigned to a specialized program at Yale for rapid training of young physicians. During his year at Yale, Motulsky completed the premedical courses that he still was lacking. More importantly, he also “took genetics with Donald Poulson and was hooked forever.”

In 1944 Motulsky served briefly as an orderly at an Army hospital near Boston, before returning to medical school at the University of Illinois, as a private first class. He was released from the Army in 1946, finished medical school in 1947, and began his residency in internal medicine at Chicago’s Michael Reese Hospital; supported by a fellowship in hematology. Under his fellowship mentor, biochemist and hematologist, Karl Singer, Motulsky began his foray into research, investigating hereditary hemoglobin disorders.

In 1951, the Korean War broke out and Motulsky was again in the Army; this time at a new hematology research unit at Walter Reed Hospital in Washington, DC. “At Walter Reed, we studied hemolytic anemias that might explain problems encountered by our soldiers… Singer had encouraged me to think about biochemical mechanisms, and here were genetic diseases, for which mechanisms could be studied to the direct benefit of patients.”

In 1953 Motulsky joined the faculty at to the University of Washington in Seattle, where he taught internal medicine and specialized courses in hematology. That same year, Watson and Crick determined the structure of DNA, rousing Motulsky to slip what he called “bootleg medical genetics” into his hematology lectures. In 1957 he founded the Division of Medical Genetics at UW; one of the first such divisions in the United States. [The Division of Medical Genetics at Johns Hopkins opened the same year.] But, before that, to broaden the scope of his knowledge, Motulsky spent eight months in the human genetics unit of Lionel Penrose at University College London. “Penrose’s department was the best in human genetics in Europe, including, in addition to Penrose himself, J.B.S. Haldane… People were very critical and helped me recognize excellent work.” Concurrently, Motulsky became interested in what is now called pharmacogenetics—”that is, differential reaction to drugs among people based on their genotypes.”

This blog post was meant to recount Motulsky’s remarkable personal journey during the early years of his life and career. Nonetheless, Motulsky’s considerable scientific contributions also need to be noted, if only briefly—in particular, his analyses of the genetic predisposition to a broad assortment of conditions, including heart disease, blood disorders, colorblindness, infections and immunity, hypertension, and alcoholism (for a concise account of his achievements, see his short autobiography referenced below). It is a good story. For now, note the comments of Francis Collins—a geneticist, and the director of the National Institutes of Health—which Collins made after Motulsky’s passing. “The relationship between heredity and the response to drug therapy — nobody was thinking about that until he started, 60 years ago. He anticipated it decades before science made it possible to get the answers that he dreamed of.” Moreover, “There were very few medical centers that considered genetics as being all that relevant to human medicine. It was more a study of fruit flies and mice, not humans.” Also note the comment of Claire King, Motulsky’s colleague at the University of Washington (and reporter of his autobiography), who discovered the role of certain genetic mutations in breast cancer. Because of Motulsky’s work in medical genetics, “the field is now integrated into every other field of medical practice and has become the soul of precision medicine.”

Motulsky ends his autobiography as follows: “Throughout my career, I have very much enjoyed the practice of medicine. Until I was 70 years old, I was an attending physician in general internal medicine. The human contact, to be able to help people, is enormously rewarding. And I always learned something. I was stimulated to ask new questions by seeing things that I hadn’t previously thought about. And with respect to medical genetics in particular, I know of no other field in science and medicine that is as fascinating. Medical genetics is closer to science than most specialties in medicine. It reveals exciting new biological phenomena. It has social implications and historical implications and ethical implications. I cannot imagine having done anything more exciting for the past 70 years.”

References 

Dallek, R. Franklin Roosevelt and American Foreign Policy, 1932-1945. Oxford University Press. 1979.

Berenbaum, M. The World Must Know: The History of the Holocaust as told in the United States National Holocaust Memorial Museum. Little, Brown and Company, 1993.

Gradyjan, D. Arno Motulsky, a Founder of Medical Genetics, Dies at 94. Obituary, N.Y. Times, January 29, 2018

Motulsky, A. G. (as told to Mary-Claire King). 2016. The Great Adventure of an American Human Geneticist. Annual Review of Genomics and Human Genetics 17:1-15. https://doi.org/10.1146/annurev-genom-083115-022528

Addendum:

Several posts on the blog feature individuals whose lives were impacted in major ways by World War II and the Holocaust. In particular:

The Life and Legacy of George Klein: Cancer Virus Pioneer and Witness to the Holocaust, May 4, 2017.

Gunter Blobel: Nobel Laureate Best known For the Signal Hypothesis of Protein Targeting, February 22, 2018

The Converging Lives of Jacques Monod, Francois Jacob, Andre Lwoff, and Albert Camus in Wartime France, March 27, 2017

 

 

Dr. Arno Motulsky in his Seattle apartment in 2014. Credit Clare McLean/University of Washington
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Gunter Blobel: Nobel Laureate Best known For the Signal Hypothesis of Protein Targeting

Günter Blobel, recipient of the 1999 Nobel Prize in Physiology or Medicine, died on February 18, 2018. Blobel is best known for his groundbreaking studies on an issue of fundamental importance to cell biology and virology—how newly synthesized proteins reach their correct location within or outside the cell.

Gunter Blobel (4)

In Blobel’s words: “Concurrently with or shortly after their synthesis on ribosomes, numerous specific proteins are unidirectionally translocated across or asymmetrically integrated into distinct cellular membranes. (1).” How this might be realized, now known as the “signal hypothesis,” was first put forward in 1971 by Blobel and colleague David Sabatini.  The central feature of the hypothesis is “the occurrence of a unique sequence of codons, located immediately to the right of the initiation codon, which is present only in those mRNAs whose translation products are to be transported across (my note: or inserted into) a membrane (2).”

Blobel and postdoc Bernhard Dobberstein (one of the impressive cell biologists trained by Blobel) confirmed the hypothesis in a classic experiment based on an earlier observation by Cesar Milstein. Specifically, Milstein’s group found that proteins, produced by translation of immunoglobin light chain mRNA, in a cell-free translation system, contained about 20 amino acids at their N-terminal end that are not present on immunoglobin light chains that are secreted from cells. Blobel and Dobberstein confirmed that cell-free translation (on free ribosomes) of immunoglobin light chain mRNA indeed yields the larger form of the protein. However, when membranes were added to the system, the light chains that were synthesized were the same size as normally secreted light chains. What’s more, the ribosomes synthesizing the light chains were bound to the membranes, and the new light chains were resistant to digestion by added proteases, indicating that they had been secreted into microsomes (2).

The overall scheme is that those nascent proteins, which are destined for secretion, or insertion into membranes, contain a short signal sequence at their N-terminus, which causes their ribosome to attach to a membrane. The signal sequence is removed by a signal peptidase as the growing polypeptide passes through a channel in the membrane. Proteins destined to be transmembrane proteins also contain a hydrophobic stop-transfer sequence, which anchors them in the membrane. Note that important details of this fundamental cellular pathway were revealed by Blobel’s analysis of the strategy by which sindbis virus generates its envelope glycoproteins (3). [For a detailed review of this key step in the replication cycles of enveloped viruses, see Chapters 7 and 8 of L. Norkin, Virology: Molecular Biology and Pathogenesis, ASM Press, 2010.]

Gunter Blobel was born in May 1936, in the Silesian village of Waltersdorf, then in eastern Germany, and later a part of Poland. He is another of the scientists featured on the blog whose life was impacted by events of the Second World War. Others include Max Delbruck, Francois Jacob, Jacques Monod, Andre Lwoff, the Wollmans (Eugene, Elizabeth, and Elie), Renato Dulbecco, Harald zur Hausen, and George Klein.

In Blobel’s words (4): “1945 was also a turning point in my life. Until then my childhood was a perfect 19th century idyll. In the cold and snow-rich Silesian winters there were hour-long rides on Sundays in horse-drawn sleighs to my maternal grandparent’s farm to have lunch and to spend the afternoon. The house was a magnificent 18th century manor house in the nearby Altgabel with a great hall that was decorated with hunting trophies. In the summer, of course, horse-drawn landauers were used as means of transportation. The way to school was a long one. We went there on foot and as a pack, usually consisting of one or two of my seven brothers and sisters and of children from neighboring houses.

At the end of January 1945, we had to flee from the advancing Russian Red Army. My father, a veterinarian stayed behind for a few more days and left only hours before the Red Army moved in. My fourteen-year-old brother, Reiner, drove my mother, my youngest brother, an older brother, the two younger sisters and me in a small automobile to relatives west of Dresden in Saxony. On the way there we drove through Dresden. We entered the city from the eastern hills. Its many spires and the magnificent cupola of the Frauenkirche (die Steinerne Glocke, the Stone Bell) were a magnificent sight even for the untrained eye of a child. Driving through Dresden, I still remember the many palaces, happily decorated with cherubs and other symbols of the baroque era. The city made an indelible impression on me. Only a few days, later, on February 13, 1945, we saw from a distance of about 30 kilometers a fire-lit, red night sky reflecting the raging firestorm that destroyed this great jewel of a city in one of the most catastrophic bombing attacks of World War II. It was a very sad and unforgettable day for me.

The months before and after the end of World War II were chaotic and miserable. None of my relatives had enough space to accommodate our large family leaving us divided among several relatives in different villages. There was no communication and little food. On September 9, 1945, we learned of the death of my beautiful oldest sister Ruth who, at age 19, was killed in an air raid on a train she was travelling in on April 10, 1945. She was buried in a mass grave near the site of the attack in Schwandorf, Bavaria. Ruth was born when my mother was just 20. The two had a sisterly relationship. My mother grieved over Ruth’s death until the end of her own life.

Fortunately, things took a turn for the better, when my father was able to continue his veterinarian practice in the charming medieval Saxon town of Freiberg. Most members of our family were reunited there by 1947. We lived in a nice villa surrounded by a large garden on the edge of town. My way to school was along the old medieval city wall. For only 40,000 inhabitants, Freiberg had a rich cultural life with a 175-year-old theater. Most impressive were the musical performances in the magnificent gothic cathedral, the Dom, with the splendid great Silbermann organ. Each week Bach cantatas were performed. The great choral works of Bach, Mozart and Haydn were regularly performed and at the highest artistic level at the major religious holidays. I even participated in singing in the cantus firmus of Bach’s Matthäus Passion. So, it was almost like a 19th century idyll again, this time in a small medieval town instead of a country village.

However, there was now the ever more oppressive regime of East Germany to deal with on a daily basis. When I graduated from high school in 1954 I was not allowed to continue my education at a university because I was considered a member of the “capitalist” classes. [My note:, Gunter was labeled “a member of the capitalist classes,” and barred from universities, for refusing to join a Communist youth group.] Fortunately, at that time, i.e., before the Berlin Wall, it was possible to escape and to travel freely to West Germany. So, on August 28, Goethe’s birthday, I left Freiberg for Frankfurt on the Main in West Germany. The train left in the morning and in the afternoon, it passed Weimar, where Goethe spent most of his life, and then Eisenach, where Bach was born and in the evening it arrived in Frankfurt, Goethe’s birthplace.”

Several paragraphs later:

“In 1994, I founded Friends of Dresden, Inc., a charitable organization, with the goal to raise funds in the U.S. to help rebuild the Frauenkirche in Dresden. The rebuilding of many of the historic monuments of Dresden is one of the most exciting consequences of German reunification and the liberation from communism. It is a childhood dream come true.

It was one of the great pleasures of my life to donate the entire sum of the Nobel Prize [my note: $960,000], in memory of my sister Ruth Blobel, to the restoration of Dresden, to the rebuilding of the Frauenkirche and the building of a new synagogue. This donation also serves to express my gratitude to my fellow Saxons. They received us with open arms when we had to flee Silesia. I spent a wonderful period of my life there and they gave me a thorough and valuable education. A few thousand dollars will also be donated for the restoration of an old baroque church in Fubine/Piemonte/ltaly, the home town of my wife’s father, Sebastanio Maioglio. We have spent many happy summers there in the parental home of my wife.” [My notes: the Frauenkirche was destroyed by the Allied bombing, and the Nazis destroyed the synagogue was in 1938. Globel also took up the rebuilding of the Paulinerkirche, the university church of the University of Leipzig, which had been blown up by the communist regime of East Germany in 1968.]

Blobel earned a degree in medicine from the University of Tubingen in 1960. However, after his internship in Germany, he decided that as a doctor he was merely treating symptoms, and that to get at causes he must turn to research. Blobel earned a doctorate in oncology from the University of Wisconsin in 1967. Next, he was a postdoctoral fellow at the Rockefeller Institute, under Nobel laureate, George Palade, and eventually became a full professor at the Institute in 1976, where he remained active until his recent death.

References:

  1. Blobel, G. (1980). Intracellular protein topogenesis, Proc Natl Acad Sci USA, 77:1496-1500.
  2. Blobel, G., and B. Dobberstein. (1975). Transfer of proteins across membranes. 1. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol, 67:835-851.
  3. Bonatti, S., R. Cancedda, and G. Blobel (1979). Membrane biogenesis. In vitro cleavage, core glycosylation, and integration into microsomal membranes of sindbis virus glycoproteins. J Cell Biol, 80:219-230. DOI: 10.1083/jcb.80.1.219
  4. “Günter Blobel – Biographical”. Nobelprize.org. Nobel Media AB 2014. Web. 21 Feb 2018. <http://www.nobelprize.org/nobel_prizes/medicine/laureates/1999/blobel-bio.html&gt;

Knowledge Gaps in the Pursuit of a Universal Influenza Vaccine

Seasonal influenza outbreaks cause between 250,000 to 500,000 deaths word-wide each year (according to 2008 WHO estimates).  What’s more, unpredictable pandemics, of which there were four in the 20th and the 21st centuries (1918, 1957, 1968, and 2009) pose a still greater threat. The worst of these pandemics, the 1918 Spanish flu outbreak, claimed an estimated 50 to 100 million lives globally (according to 2014 WHO estimates).

The human population was “only” 1.9 billion individuals during the 1918 pandemic, whereas there now are about 8 billion people inhabiting our planet. Thus, a future pandemic might be far more catastrophic than the 1918 episode. Moreover, since pandemic strains are derived in part from zoonotic influenza viruses, the constant rise in livestock numbers, intensive farming, and the numbers of animals being transported around the world, combine to facilitate the genetic mixing and evolution of influenza viruses, and the chance of an animal influenza virus becoming able to jump to humans and causing a pandemic.

Unlike vaccines against other viruses (e.g., measles), the seasonal flu vaccine needs to be updated each year to keep up with the antigenic changes that continually occur in influenza viruses. This creates several problems, the first of which is that individuals need to be re-vaccinated each year. And, since it takes months to produce a vaccine, when the updated vaccine is at last ready, it may not be a particularly good match against the new season’s strains. The current vaccine is only about 30 percent effective, which accounts at least in part for the unusually severe flu season we are currently experiencing. And while the efficacy of the current vaccine may be atypically low, even in good years the match is less than optimal.

Pandemics present a much greater challenge to vaccine makers, since pandemics may be vastly more severe than seasonal outbreaks, and since an entirely new vaccine is needed against pandemic viruses. That latter is so because, as noted, pandemic strains are derived in part from zoonotic influenza viruses, which by-and-large are antigenically distinct from strains already circulating in the human population, that humans already express immunity against. Consider the example of the 2009 pandemic. Because an entirely new vaccine was needed to meet the threat of the pandemic virus, the vaccine was not available until after the first wave of infection had already occurred. Fortunately, the 2009 pandemic virus was relatively mild.

A “universal” flu vaccine, that could provide lifelong protection against all seasonal strains of influenza, as well as provide protection against a pandemic virus, would be a most crucial and significant breakthrough. A major international workshop, entitled “Pathway to a Universal Influenza Vaccine,” was convened June 28 and 29, 2017, by the U.S. National Institute of Allergy and Infectious Diseases, to identify gaps in our knowledge that need to be addressed to develop such a vaccine (1).

Participants noted shortcomings in our understanding of the epidemiology, transmission, natural history, and pathogenesis of influenza. Among the issues specifically mentioned: “influenza surveillance is lacking in certain regions of the developing world and globally in certain high-risk groups… gaps in knowledge include the relationship between symptoms, viral shedding and transmission, as well as the level of protection needed to interrupt transmission.”

The host factors that influence influenza disease severity were also acknowledged to be poorly understood. To that point, participants addressed the need to better understand how pre-existing immunity—which might result from multiple natural influenza infections, as well as from repeated vaccinations—might affect how that person’s immune response will respond to future infections and, importantly, how past exposures might affect the efficacy of a vaccine. As stated in the meeting report: “Recent data provide strong epidemiologic evidence that infection with the influenza strain circulating during one’s childhood elicits a lifelong immunologic imprint that impacts responses to novel strains and can help protect against unfamiliar HA subtypes from the same phylogenetic group as the original infecting virus…The potential consequences of imprinting infants with vaccines versus natural exposure need to be carefully assessed.” [The HA protein is the so-called hemagglutinin, which serves the virus as its attachment protein.]

Participants also noted gaps in our understanding of the underlying B and T cell immune mechanisms that are induced by both natural infections and vaccinations. Further study of these responses was recommended so that we might be better able to stimulate them.

As might be expected, it is singularly important to identify the antigens that might be the most promising targets of a universal vaccine. To that point: “…most areas of the HA head are subject to antigenic change, and therefore unlikely to yield a broadly protective immune response. Neutralizing antibody responses to conserved regions such as the HA stalk, and non-neutralizing antibodies such as those directed at the neuraminidase (NA), and matrix 2 ectodomain, merit further study…The importance of each site may differ for pandemic versus seasonal influenza.” [The HA head region binds to receptors on the cell surface. After the bound virus is taken into the cell by receptor-mediated endocytosis, the low pH within endosomes triggers a conformational rearrangement of the HA stalk region, which mediates fusion of the viral envelope with the endosomal membrane, thereby releasing the viral cores into the cytosol. Since mutations within the stalk region might disrupt its membrane fusion function, such mutations are generally selected against. Researchers have recently had success developing antibodies that target the neuraminidase protein at the viral surface.]

Participants acknowledged that animal models play an important role in influenza research, especially when studying pandemic viruses. Nonetheless, “Animal models have limitations including the inability to mimic the human experience regarding genetic background, lifetime exposure to natural influenza infection or vaccine, viral susceptibility…and determinants of immune response and protection.” Thus, the participants noted that “a human a challenge model will be a crucial tool for vaccine development, as it can help answer fundamental questions about influenza immunity and serve as a mechanism for rapidly testing the efficacy of new products…Expansion of this resource should be a top priority…”

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Day two of the workshop consisted of a rapporteur session on key conclusions, chaired by David Baltimore and Anthony S. Fauci (1)

Participants agreed that a robust collaboration between government agencies, academia, and industry would be needed to translate the fruits of basic research into a universal influenza vaccine. To that point, a January 24, 2018 editorial in Nature (doi: 10.1038/d41586-018-01070-w) asserted: “…advocates rightly argue that the research and development of a universal flu vaccine — ultimately the only effective defense against future pandemics — merits a program equivalent in scale to the Manhattan Project.” Yet the US government last year invested just $75 million on universal flu vaccine research and development.

    1. Paules C.I., D. Marston, R. W. Eisinger, D. Baltimore, and A. S.Fauci. (2017), Meeting Report: The Pathway to a Universal Influenza Vaccine. Immunity 47: 599-603. https://doi.org/10.1016/j.immuni.2017.09.007

 

 

Douglas Lowy, John Schiller, and the Vaccine Against Cervical Cancer: Postscript

Our October 12, 2017 post, Douglas Lowy, John Schiller, and the Vaccine Against Cervical Cancer, has reached a gratifying number of people. Since some readers might welcome a bit more background vis-à-vis the remarkable human papillomavirus (HPV) life cycle, or details concerning the use of virus-like particles (VLPs) in the experimental stages of the vaccine’s development, or how the vaccine might actually work, here are a few additional points.

The post noted that the replication cycle of HPV is regulated by the differentiation states of the cells making up the layers of an intact, stratified epithelium or mucosae. “Since the outer layer of the skin is comprised of dead cells, cutaneous HPV infection requires a break or puncture of the skin for the virus to access cells of the underlying germinal stratum of the epithelium. In the actively dividing basal cells, the viral genome replicates more frequently than the cellular genome, thus amplifying the viral genome copy number. However, because the viral genes that encode the capsid proteins are not expressed in these cells, progeny virus particles, which might induce an immune response, are not yet produced. As the basal cells differentiate and move up in the epithelium, the viral genomes replicate only once per cell cycle, on average, to maintain the viral genome copy number. Then, as the infected cells go through their final stages of differentiation in the outer layers of the epithelium, the virus life cycle switches to its productive phase. Capsid proteins are produced, and thousands of virus particles are generated from the each of the infected, terminally differentiated cells.” [How cellular differentiation regulates HPV gene expression and replication is detailed in the textbook, Virology: Molecular Biology and Pathogenesis.]

The post noted that by coupling its replication cycle to the differentiated state of the host cell within the stratified epithelium, HPV can produce progeny virus particles only in the terminally differentiated cells that comprise the outermost live cells of the epithelium. In this way, HPV productive infection does not activate an antiviral immune response. [The host’s immune response eventually does clear many HPV infections. Also, the incidence of HPV-associated lesions is higher in immunosuppressed patients.]

Here then is an additional key point. After the amplification stage, the viral genomes replicate in the basal cells, but only in conjunction with cellular DNA replication. In that way, the viral genome copy number is maintained in the basal cells. Moreover, and importantly, when the basal cells divide, one daughter cell remains behind as a basal cell, while the other daughter cell migrates up into the epithelium. Thus, one daughter cell will differentiate and thereby enable the virus to complete its replication cycle—at a level in the epithelium or mucosae beyond the reach of immune attack—while the other daughter cell remains behind in the basal layer, where it sustains the persistent infection.

Another consequence of this remarkable replication cycle is as follows. Since there are no blood or lymphatic vessels in the stratum of the epithelium or mucosae where the productive replication is occurring, the infection tends to remain localized, thereby giving rise to warts or tumors.

Since HPVs are difficult to study and propagate, one might ask how Lowy and Schiller were able to assess the antibody titers that were induced by inoculation with the HPV VLPs. The answer is that they used a pseudovirion-based immune assay. Pseudovirions are essentially VLPs that contain a plasmid that carries a reporter gene.

One last point. I believe it is generally the case that vaccines due not prevent virus infections per se. Rather, they enable the host to bring an infection under control more quickly, before symptoms might arise. Considering that cervical carcinomas may develop after years of virus persistence, despite a continuing immune response against the virus the whole time, how then might the vaccine protect against the cancer? Here is a thought. Bearing in mind that the human immune response naturally clears many HPV infections over time, perhaps the vaccine protects the host by enhancing immune surveillance to clear the infection before the emergence, or malignant progression of HPV-induced lesions. Or, perhaps the vaccine actually prevents infection.

 

 

 

Douglas Lowy, John Schiller, and the Vaccine Against Cervical Cancer

The 2017 Lasker-DeBakey Prize for Clinical Research went to two virologists at the National Cancer Institute, Douglas Lowy, 75, and John Schiller, 64, for developing technologies that led to FDA-approved vaccines against human papillomavirus (HPV) strains that cause cervical carcinoma and other cancers.  Lasker awards are considered the United States’ most prestigious biomedical research awards. They often precede a Nobel Prize in Physiology or Medicine. Thus, they are referred to as “America’s Nobels.” Eighty-seven Lasker awardees have gone on to win a Nobel.

Douglas Lowy and John Schiller

Lowy and Schiller’s achievements were prompted by Harald zur Hausen’s 1983 discovery that two HPV subtypes, HPV-16 and HPV-18, together account for about 70% of all cervical cancers. Since more than 120 distinct HPV subtypes had been identified, the high degree of association of cervical carcinoma with only two of these subtypes provided compelling evidence for the viral etiology of cervical carcinoma. Later studies showed that HPV-31, HPV-33, HPV-45, HPV-52, and HPV-58 are responsible for another 20% of cervical cancers. Thus, an HPV infection can be detected in virtually all cervical carcinomas. Harald zur Hausen was awarded a share of the 2008 Nobel Prize in Physiology or Medicine for his discovery. [His story is told in Harald zur Hausen, Papillomaviruses, and Cervical Cancer, posted June 19, 2015.]

Lowy and Schiller did not begin their work on papillomaviruses with the intent to produce a vaccine. Instead, like many papillomavirus researchers at the time, they were investigating how papillomavirus oncogene products affected cell growth and replication (i.e., how they cause cancer). Toward that end, they were making use of bovine papilloma virus (BPV) in their studies, rather than HPV. BPV was easier to work with than HPV, because BPV, but not HPV, could be studied in standard cell cultures (see Aside 1).

[Aside 1: The replication cycle of HPV depends upon the differentiation states of the cells making up the layers of an intact, stratified epithelium. Details are as follow. Since the outer layer of the skin is comprised of dead cells, cutaneous HPV infection requires a break or puncture of the skin for the virus to access cells of the underlying germinal stratum of the epithelium. In the actively dividing basal cells, the viral genome replicates more frequently than the cellular genome, thus amplifying the viral genome copy number. However, because the viral genes that encode the capsid proteins are not expressed in these cells, progeny virus particles, which might induce an immune response, are not yet produced. As the basal cells differentiate and move up in the epithelium, the viral genomes replicate only once per cell cycle, on average, to maintain the viral genome copy number. Then, as the infected cells go through their final stages of differentiation in the outer layers of the epithelium, the virus life cycle switches to its productive phase. Capsid proteins are produced, and thousands of virus particles are generated from the each of the infected, terminally differentiated cells. Thus, the HPV life cycle is regulated by the differentiated state of the host cell within the stratified epithelium. Because virus production is restricted to the outermost layers of the epithelium, the virus can evade the immune system, such that the infection can persist, and be passed on for years. However, in most instances, the host appears to eventually mount a successful immune response, which clears the infection.

The development of so-called organotypic raft cultures eventually made it possible to study HPV in cell culture. But one could produce only very limited amounts of the virus in that system.]

Working with BPV, Lowy and Schiller developed protocols they would later use when they turned their attention towards an HPV vaccine. One of these protocols was for an assay to measure the titer of neutralizing antibodies against BPV. Importantly, they also discovered that they could generate “virus-like particles” (VLPs), comprised only of the major BPV coat protein (L1). The BPV L1 proteins (which were generated by a baculovirus vector in insect cells) self-assembled into VLPs that were morphologically like actual BPV particles. What’s more, using their assay to measure the titer of neutralizing serum antibodies, they found that the VLPs induced neutralizing antibodies in rabbits that were effective against the actual virus. Importantly, since the VLPs did not contain viral genes, they could not cause cancer.

Again, using their assay for measuring the titer of neutralizing antibodies against BPV, Lowy and Schiller compared the immunogenicity of BVP VLPs, to that of individual BPV proteins. The VLPs indeed are more immunogenic than individual viral proteins, since they induced higher levels of neutralizing antibodies than were induced by individual L1 proteins (see Aside 2).

[Aside 2: The activation of antibody-producing B-cells is triggered by the cross-linking of their antigen-binding B-cell receptors, which is facilitated by the multimeric VLPs, but not by individual viral proteins.]

The innovations resulting from their work with BPV would enable Lowy and Schiller to overcome the formidable challenges they faced when working to develop the HPV vaccine. One obstacle was that HPV cannot replicate in standard cell cultures. Thus, it was difficult to study HPV, and importantly, it also was difficult to propagate it. Being able to propagate substantial amounts of the virus would be necessary to produce a vaccine.

Another obstacle to an HPV vaccine was the potentially unacceptable risk of inoculating people with a virus (either attenuated or killed) that contains known oncogenes. Lowy and Schiller overcame this impediment, and the one noted above, by implementing protocols they previously developed while researching BPV. Specifically, they generated HPV VLPs that were comprised only of the HPV L1 capsid protein, and which induced an immune response that produced protective antibodies. [They used the L1 protein of HPV-16; the most carcinogenic strain of HPV.]  In addition, they developed cell lines, which contained high copy numbers of the plasmid that encoded the HPV L1 protein; a step which enabled them to scale-up production of the VLPs.

Together, these breakthroughs made a compelling case for the feasibility of an HPV vaccine. So, Lowy and Schiller prevailed upon several pharmaceutical companies to produce a vaccine in commercial amounts, and to see the vaccine through the clinical trials process. Most companies remained skeptical about the ultimate success of the vaccine. But two companies, Merck and GlaxoSmithKline (which later bought Merck), accepted the challenge. Thus, Merck developed Gardasil, while GlaxoSmithKline developed Cervarix. [The VLPs in Gardasil are produced in yeast, whereas the VLPs from Cervarix are produced in insect cells, via a recombinant baculovirus.]

Clinical trials showed that the Merck and the GlaxoSmithKline vaccines induce significant antibody titers against high-risk HPVs. The US FDA approved the respective HPV vaccines in 2006 and 2009.

The HPV vaccines have had a substantial effect on human health. Consider the following: Cervical cancer is the second most common cause of death from cancer among women worldwide. HPV infection is the cause of virtually all cases of cervical cancer. HPVs also cause 95% of anal cancers, 70% of oropharyngeal cancers (more common in men than in women), 65% of vaginal cancers, 50% of vulvar cancers, and 35% of penile cancers. Next, consider that, since Gardasil and Cervarix were introduced, HPV infection rates have dropped by 50 percent among teen-age girls in U.S., even though only a third of teens between 13 to 17 years-old have received the full course (3 shots) of the vaccine (see Aside 3).

[Aside 3: Current CDC recommendations are as follows: “All kids who are 11 or 12 years old should get two shots of HPV vaccine six to twelve months apart. Adolescents who receive their two shots less than five months apart will require a third dose of HPV vaccine…If your teen hasn’t gotten the vaccine yet, talk to their doctor or nurse about getting it for them as soon as possible. If your child is older than 14 years, three shots will need to be given over 6 months. Also, three doses are still recommended for people with certain immunocompromising conditions aged 9 through 26 years.”]

Although he HPV vaccines have significantly reduced the incidence of cervical cancer in the developed world, the rates of cervical cancer in the United States are needlessly high, in comparison to the rates in other industrialized nations. The HPV vaccines have a lower acceptance rate than other childhood vaccines in the United States, perhaps because many American parents, some of whom associate with the religious right, have reservations about vaccinating their children against a sexually transmitted disease. Other individuals, liberals as well as conservatives, may oppose vaccines in general because they distrust pharmaceutical companies, or because they resent government interference in their lives. In any case, the CDC found no evidence of any increase in sexual activity among teenage girls who received the vaccine. Nor did it report any major ill effects]. See Aside 4.

[Aside 4: Since HPVs alone account for about 5% of all human cancers worldwide, we might ask what percentage of human cancers have a viral etiology. Hepatitis C virus, a flavivirus, and hepatitis B virus, a hepadnavirus, cause hepatocellular carcinoma; Epstein-Barr virus (EBV), a herpesvirus, causes Burkitt’s lymphoma and nasopharyngeal carcinoma; human herpesvirus 8 (HHV-8), causes Kaposi’s sarcoma, the most frequent cancer seen in AIDS patients; the human T-lymphotropic retrovirus I (HTLV-I) induces adult T-cell leukemia; and Merkel cell polyomavirus (MCV) causes its eponymous cancer. Together, viruses may account for as many as 20% of all human cancers, and a similar percentage of all deaths due to cancer!

As shown by the HPV vaccine, and earlier by vaccines against hepatitis B, cancers that have a viral etiology might be prevented by vaccination. Apropos hepatitis B, in the late 1980s, Merck and GlaxoSmithKline developed the respective hepatitis B vaccines, Recombivax and Engerix. Like, the HPV vaccines, they are based on VLPs, and they have significantly reduced the incidence of HBV-associated hepatoma; once one of the most lethal cancers.

Bacterial and parasitic infections too may lead to cancer. For example, Heliobacter pylori infections may lead to stomach cancer, and Schistosoma, Opisthorchis, and Clonorchis have been linked to rectum and bladder cancers in areas of Northern Africa and Southeast Asia, where those pathogens are prevalent.]

Lowy and Schiller’s achievement stands out as a superb example of basic research translating into very considerable public health benefits. Moreover, it serves as a strong endorsement for government support of basic research. To these points, Schiller noted that companies would not likely have carried out the necessary basic research and development necessary to produce the HPV vaccine, considering the seemingly small likelihood of success, as suggested by earlier failed attempts to develop a vaccine.

At a September 6, 2017 press conference announcing the Lasker-DeBakey Clinical Medical Research Award, Lowry related that he first learned about vaccines in 1955, when he went with his mother, a physician, to a talk by Jonas Salk about his then new polio vaccine. “I learned far more about polio virus and the vaccine than was probably appropriate for a 12-year-old boy.” Many years afterwards, Lowy began his “extraordinarily effective” collaboration with Schiller, which has endured for more than 30 years.

Schiller said that a high point in his career was taking his daughter to get the vaccine he helped to create. “We first came up with the idea of the vaccine when she was born and it became available when she was 13 years old (1).”

References:

  1. A Revolutionary Vaccine, New York Times, September 6, 2017.

 

 

 

 

 

 

Thomas Jefferson: Fighting Smallpox

Variolation was the world’s first practical measure to control smallpox. It was developed in China in the 11th century. The procedure involved inoculating uninfected individuals with material from the scabs of individuals who survived smallpox infection.

It was brought to England for the first time in 1721, by Lady Mary Wortley Montague—the wife of the British ambassador to Turkey—when she returned home after learning of the practice in Istanbul. It was brought to Colonial North America the same year by the prominent Puritan minister, Cotton Mather. New England was then experiencing a major smallpox epidemic.

Mather is perhaps best known for his role in the Salem witchcraft trials. He learned of variolation not from the British, but from his African slave, Onesimus, who had been inoculated as child in Africa. Onesimus was a “gift” to Mather in 1706, from his Boston congregation. Variolation was used in western Africa when Onesimus was a child.  The practice may have been brought there by caravans from Arabia. In any case, an enslaved African man played a key role in bringing variolation to North America.

In 1777, during the American War for Independence, General George Washington required the entire Continental Army to undergo variolation. Bearing in mind that more than two-thirds of the American casualties during the War resulted from disease, and that smallpox alone caused a total of about 100,00 deaths, some historians maintain that Washington’s policy of enforced variolation was his most important strategic decision of his entire military career.

Variolation nonetheless encompassed risks—a fatality rate of 1 to 2%—that would be unacceptable today. Not surprisingly then, the colonial and Revolutionary War periods were times when public fear and restrictive laws often prevented the use of variolation. Nonetheless, Thomas Jefferson was a lifelong advocate of smallpox-prevention measures. In 1766, Jefferson traveled to Philadelphia to undergo variolation, since the practice was banned in his native Virginia. As a lawyer in 1768, Jefferson defended a Norfolk doctor, whose house was burned down by a mob because he practiced variolation. In 1769, Jefferson placed a bill before the Virginia General Assembly to reduce the 1769 restrictions against variolation. In the 1770s and 1780s, he had his children and his enslaved servants (including Sally Hemings, his wife’s half-sister, and mother of several of his enslaved children) undergo the procedure.

In 1799, Boston physician and founder of Harvard Medical School, Benjamin Waterhouse, introduced Edward Jenner’s new cowpox-based smallpox vaccine to New England. Wanting to spread word of the vaccine to the rest of the new country, in 1781 Waterhouse sent a sample to his friend, Thomas Jefferson. At the President’s House in Washington, Jefferson selected an enslaved kitchen worker to be the first recipient of the vaccine. However, the vaccine did not take. So, Jefferson then had two of his slaves at Monticello undergo vaccination. When those vaccinations proved to be successful (as shown by exposure to actual smallpox), Jefferson serially transmitted the vaccine from the two original vaccines to almost fifty other slaves. By means of serial inoculations he then sent vaccine material to Washington (the city), and from there the vaccine traveled to Philadelphia and beyond. So, Thomas Jefferson, and his African slaves, played a seminal role in protecting many people in the new United States from smallpox.

Jefferson was an amateur, but serious scientist. He kept detailed notes of his observations, and corresponded with Jenner. Here is what he wrote to Waterhouse about the appearance of papules at the vaccination site:

As far as my observation went, the most premature cases presented a pellucid liquor the sixth day, which continued in that form the sixth, seventh, and eighth days, when it began to thicken, appear yellowish, and to be environed with inflammation. The most tardy cases offered matter on the eighth day, which continued thin and limpid the eighth, ninth, and tenth days. [http://www.smithsonianmag.com/smart-news/thomas-jefferson-conducted-early-smallpox-vaccine-trials-180954146/]

 

 

 

 

Julius Youngner: Slighted Polio Vaccine Pioneer

This is a tale of the hurt that a junior investigator might feel when a senior investigator takes the lion’s share of the credit for the junior investigator’s crucial breakthroughs. Jonas Salk, who conceived and oversaw the development of the first widely used polio vaccine, is the senior investigator in this anecdote. Julius Youngner, the last surviving member of the original vaccine research team that Salk assembled in the early 1950s at the University of Pittsburgh, is the slighted assistant. Youngner later had his own distinguished career. He passed away in April of this year. Here is their story.

After earning his Ph.D. in microbiology, Youngner was drafted into the World War II U.S. Army, which assigned him to the Manhattan Project, to test the toxicity of uranium salts. Youngner first learned the purpose of the Manhattan Project when the first atomic bomb was dropped on Japan.

After the war, Youngner worked as a commissioned officer for the U.S. Public Health Service. This was a significant stop in his career, since it was there that he first became interested in viruses and cell culture. But, since there was no opportunity for him to pursue that interest in Bethesda, he began to look elsewhere. Thus, it happened in 1949 that Salk recruited Youngner to join his vaccine research team in Pittsburgh, after a mutual acquaintance told Salk that Youngner was eager to work on viruses and cell culture.

Jonas Salk and Julius Youngner at the University of Pittsburgh, early 1950s

Salk hoped that Youngner might find a way to generate enough cells from monkey kidney tissue to support mass-production of the vaccine. Youngner, on his own, then developed the use of the proteolytic enzyme, trypsin, to disperse tissue fragments into individual cells, thereby generating many more cells from a given amount of tissue. Indeed, Youngner could generate enough cells to support manufacture of the vaccine. This was his first key contribution to the vaccine project. “Trypsinization” remains a mainstay of modern cell culture.

Youngner’s next major contribution to the vaccine enterprise was his development of a rapid analytical test that had two crucial applications. First, recalling that the Salk vaccine contains an inactivated virus, Youngner’s so-called “color test” made it possible to quickly screen batches of the vaccine for any live virus that might have survived the inactivation process.  Second, Youngner’s test made it possible to quickly test the vaccine’s ability to induce anti-poliovirus antibodies (1). [Youngner based his color test on an earlier observation by John Enders, Tom Weller, and Fred Robbins, that metabolic activity (as indicated by a drop in pH) was less in cultures inoculated with live virus than in control cultures (2, 3). In Youngner’s test, a color change of phenol red, resulting from a shift in pH, served as an indicator of virus activity, or of antibody activity.]

Some sources credit Youngner with having devised the process for inactivating the virus. But, that is correct in a very limited sense only. Salk selected incubation in formalin as the means to disable the virus. In truth, Salk learned of that approach a decade earlier while doing postgraduate studies under Thomas Francis at the University of Michigan. Francis was then using formaldehyde to produce his killed influenza vaccine (2).

What’s more, Salk’s choice of formalin to generate his polio vaccine was bold. Earlier, in the 1930s, Canadian scientist Maurice Brodie tested a formalin-killed polio vaccine in twelve children, with disastrous results. Several of the children developed paralytic poliomyelitis (4).

Clearly, too little exposure to formalin could leave enough live virus to cause paralytic poliomyelitis or death. On the other hand, too much exposure could so badly damage the virus’ proteins that they might no longer induce an immune response against the live virus. Brodie did not have analytical procedures to ensure that he had inactivated his vaccine to safe levels. In contrast, it was clear to Salk that getting the correct balance would be vital to his vaccine project, and Youngner’s color test was the means for doing so. Youngner used his test to determine that six days of incubation in a 1:4,000 formalin solution would result in one live virus particle in 100 million doses of the vaccine (5).

Since Youngner’s inactivation curve was based on only a few data points, and since it was likely that the slope of the curve might flatten out after a time, Salk added a margin of safety of six extra days. Thus produced, the vaccine induced antibody production in monkeys, while showing no signs of causing paralysis or other problems.

By 1954, 800,000 children had been successfully immunized against polio in the first clinical trial of the vaccine. In April 1955, the outcome of the trial would be announced to a very grateful public.

By 1957, Salk’s vaccine team at Pittsburgh was no longer needed, and was dispersing. Salk was making plans to leave Pittsburgh for California, where he would found the prestigious Salk Institute. Youngner, now 34 years-old, remained at Pittsburgh, where he would begin his own distinguished career.

Although Youngner was now independent of Salk, he remained bitter over his former boss’s failure to acknowledge the underlings who had labored so diligently behind the scenes to bring the vaccine to fruition. “The first rule we learned was to call him ‘Dr Salk,’ never Jonas. He would speak to us through a wall of notes and memos…Here was a guy who could always find an hour to brief some reporter at the local Chinese restaurant, but could never find the time to sit down with his own people (6).”

Youngner was particularly appalled by events involving the paper he wrote describing his color test. “After I had what I considered to be a good draft…I gave my copy to Jonas for his comments. It should be noted this was 1954, the pre-Xerox, pre-word-processing era. I had made a working transcript of the paper for my own use and it was this copy that I handed to him. Also, it should be noted that the title page had the authors listed as ‘J.S. Youngner and E.N. Ward (6).’” Elsie Ward, who served as Youngner’s technician, was a zoologist who specialized in growing viruses.

Salk intended to read Youngner’s manuscript while away on a trip.  When Salk returned a week later, he claimed that he had lost the manuscript, but that he had jotted down some notes from which he was able to produce a draft of his own. Youngner was rather incredulous that a person as meticulous and disciplined as Salk could lose such an important manuscript. Youngner’s skepticism was further roused by the fact that Salk’s version contained all the data in Youngner’s original manuscript. Salk explained that incongruity, alleging that he found Youngner’s tables, but not the text.

In any case, Youngner was especially upset by a specific change Salk made to the title page of the manuscript: “The authors were now ‘Jonas E. Salk, J.S. Youngner, and Elsie N. Ward.’ When I (Youngner) questioned the change, Jonas said that since he had to reconstruct the whole paper it was only fair that his name go first…It was obvious to me then, and is more so now, that he considered the advance in this paper a major one and he wanted his name associated with it, even though at the time he had done nothing in the lab (no kidding!) or of an advisory nature to initiate or carry out the work (6).”

Youngner could grudgingly accept that project leaders often used their senior position to appear as co-authors, or even principal authors, on papers emanating from their labs, even if their contributions were minimal. What troubled Youngner in this instance was not that Salk pulled rank, but rather his seeming duplicity.

In yet another instance—the 1955 public announcement of the successful outcome of the clinical trial—Youngner again sensed “a pattern of deception on Salk’s part to take undue credit for the discoveries of others (6).” Salk advocated for the announcement to happen at the University of Pittsburgh. However, the National Foundation for Infantile Paralysis (better known as the “March of Dimes”), which funded the vaccine project, chose the University of Michigan in Ann Arbor as the site for the announcement. That was where Michigan professor Thomas Francis supervised the evaluation of the field trial. [Note that the NIH was not able to fund research back then the way it can today. Thus, the polio vaccine project was supported nearly entirely by private donations to the National Foundation.]

Thomas Francis spoke first. Then, when Salk spoke, he acknowledged the more prominent players in the vaccine project, including Thomas Francis, Harry Weaver (director of research at the National Foundation), Tom Rivers (chairman of the advisory committees on research and vaccines for the  National Foundation), and Basil O’Connor (law partner of Franklin Roosevelt, recruited by Roosevelt in 1928 to raise funds for polio patients at Roosevelt’s Warm Springs Foundation, and a co-founder with Roosevelt of the National Foundation in 1938; (2)). Salk then acknowledged various deans and trustees at the University of Pittsburgh. Yet, he made no mention whatsoever of his dedicated coworkers in his laboratory. They had been expecting at least some recognition from their boss.

Some of Salk’s defenders argued that Salk had acted in the best scientific tradition by prefacing his printed remarks with the phrase, “From the Staff of the Virus Laboratory by Jonas E. Salk, M.D.” But, this was small consolation to Youngner and others of Salk’s coworkers, who expected to be individually acknowledged for their exhausting work on behalf of the life-saving vaccine. Indeed, they felt betrayed.

At any rate, the 1955 announcement of the success of the polio vaccine field trials was joyously received by the public. And while Youngner remained embittered over Salk’s slighting of his coworkers, he nonetheless understood that from the point of view of the National Foundation, “it was much easier to continue raising money when you have a hero, and they had an enormous public relations department that took up Jonas’ name as the hero, which he deserved…But in the meantime, Jonas was, how shall I say, not very generous to his colleagues and he made sure that nobody else was ever mentioned (6).”

The following excerpt is from Polio: An American Story (6). “In September 1963, Salk returned to Pittsburgh to attend the unveiling of his portrait in the auditorium of the University’s medical complex, a stone’s throw from the hospital where he had done his historic polio research. Before the ceremony, Salk told Dean George Bernier that he wished to speak privately with his former assistant, Julius Youngner, now a distinguished professor at the school of medicine. The two men hadn’t talked or crossed paths since Salk’s move to California in 1961. Salk saw the meeting as a courtesy to the only remaining member of his laboratory staff; Youngner had a different agenda. Speaking softly, he recalled, he slowly released the ‘hurt’ he had bottled up for more than thirty years. ‘Do you still have the speech you gave in Ann Arbor in1955? Have you ever reread it?’ Youngner began. ‘We were in the audience, your closest colleagues and devoted associates, who worked hard and faithfully for the same goal that you desired…Do you remember who you mentioned and who you left out? Do you realize how devastated we were at that moment and ever afterward when you persisted in making your coworkers invisible? Do you know what I’m saying,’ I asked. He answered that he did…Jonas was clearly shaken by these memories and offered little response.’…The two men engaged in some uncomfortable small talk before Dean Bernier returned to escort them to the ceremony. Speaking later to a reporter, Youngner admitted, ‘I got a lot of things off my chest. I’m beyond the point where I pull my punches with him. I think it was the first time he ever heard it so graphically.’ Asked if he had any regrets about working for Salk, Youngner replied: ‘Absolutely not. You can’t imagine what a thrill that gave me. My only regret is that he disappointed me.”’

Epilogue:

Jonas Salk is deservedly celebrated for developing the killed polio vaccine. That vaccine, together with Albert Sabin’s live attenuated vaccine, which followed soon afterwards, has nearly eradicated polio worldwide. Importantly, Sabin and other polio researchers believed that only a live vaccine could induce a level of immunity sufficient to protect against a challenge with live virulent virus. Nonetheless, Salk persevered in his conviction that a killed vaccine could protect against polio, and he was right.

Salk founded the prestigious Salk Institute in 1963. Yet he never himself made another notable contribution to science.

Youngner may be best known for his work on the Salk vaccine. Yet he had a distinguished career of his own at the University of Pittsburgh after Salk left. Youngner is especially noted for his contributions to interferon research. These include his finding that non-viral agents could trigger interferon induction in animals. And, in collaboration with colleague Samuel Salvin, he identified a second type of interferon, now known as gamma-interferon. Youngner also helped to explain the antiviral-effect of interferon, and he was the first researcher to demonstrate that some viruses express countermeasures against interferon.

Youngner also made important findings in the area of persistent virus infections. Importantly, he demonstrated that defective viral variants, including temperature-sensitive mutants, can play a role in the establishment and maintenance of viral persistence; doing so by impairing (modulating) the replication of the wild-type parental viruses. Based on that principle, Youngner sought to develop dominant-negative mutants of influenza virus as a novel means of anti-influenza therapy. In addition, Youngner and colleague Patricia Dowling developed a novel live attenuated vaccine against equine influenza virus, based on a cold-adapted influenza virus, which can replicate only at the temperatures found in the respiratory tract. That live vaccine was the first to prevent a serious respiratory disease of horses.

Julius Youngner, 2010

References:

  1. Salk, J.E., Youngner, J.S, Ward, E.N. (1954). Use of Color Change of Phenol Red as the Indicator in Titrating Poliomyelitis Virus or Its Antibody in a Tissue–Culture System,” American Journal of Epidemiology. 60: 214–230.
  2. Jonas Salk and Albert Sabin: One of the Great Rivalries of Medical Science, Posed on the blog March 27, 2014.
  3. John Enders: “The Father of Modern Vaccines,” Posted on the blog August 4, 2016.
  4. Vaccine Research using Children, Posted on the blog, July 7, 2016.
  5. Williams, G., Paralysed with Fear: The Story of Polio, Palgrave Macmillan, 2013.
  6. Oshinsky, D.M., Polio: An American Story, Oxford University Press, 2005.

The Life and Legacy of George Klein: Cancer Virus Pioneer and Witness to the Holocaust

George Klein, professor emeritus of tumor biology at the Karolinska Institute in Stockholm, where he worked with his wife Eva from the very beginning, passed away on December 10, 2016, at the age of 91. Klein was best known for discovering that Epstein-Barr virus (EBV)—the herpesvirus now known to cause infectious mononucleosis—causes two human cancers, Burkitt’s lymphoma and nasopharyngeal carcinoma. Moreover, Klein discovered that EBV triggers Burkitt’s lymphoma by facilitating a chromosomal translocation of the cellular c-myc oncogene, resulting in its constitutive expression. Klein also played pioneering roles in developing the concept of tumor-suppressor genes, and in opening the field of tumor immunology. Klein’s key discoveries are summarized below. But, first, Klein, like several other protagonists in these tales, was profoundly affected by events of the Second World War, and by the early days of the Cold War that followed.

From an announcement for a 2015 symposium at the Karolinska Institute, honoring George and Eva on the occasion of their 90th birthdays

George Klein’s Jewish family moved from Eastern Slovakia to Budapest in 1930. Nineteen-year-old George was working as an assistant secretary to the Jewish Council in Budapest when Nazi Germany began its occupation of Hungary in March 1944. Because George had been working for the Jewish Council, in April 1944 he chanced that to see the Vrba-Wetzler Report, known at the time as the “Auschwitz Report.” It was written by, and was secretly transmitted to the Jewish Council by Rudolf Vrba and Alfred Wetzler, two escapees from Auschwitz. It described firsthand the fate of Jews arriving at Auschwitz, and was meant to warn Hungary’s Jews, so that they might hide from, or rebel against their Nazi oppressors.

The Auschwitz report was not publicized in Hungary for reasons explained below. However, George’s supervisor at the Jewish Council gave him permission to tell his relatives and friends of what the report revealed. But they, like most Hungarian Jews, could not believe that such atrocities could actually be taking place. [During May, June, and July 1944, 437,000 Hungarian Jews were deported to Auschwitz; to be “resettled” according to the Nazis. But, in fact, most were murdered in the gas chambers.]

Klein was arrested and pressed into forced labor by the Nazis. Afterwards, since he knew the contents of the Auschwitz Report, he fled when he was about to be ordered to board one of the deportation trains to Auschwitz. Having escaped from almost certain death, he lived underground until January 1945, when the Russian Army liberated Budapest.

Forty-three years later, Klein was watching, Shoa, the monumental (nine-hour-long) French documentary film about the holocaust. Watching the movie, Klein chanced to see a man named Vrba (one of the six principal holocaust witnesses in the film) describe his experiences as a prisoner in Auschwitz. The events that Vrba recounted horrified Klein.

Later in the film, as Vrba described his escape from Auschwitz, Klein suddenly realized, “the report I had been given to read under a promise of secrecy in Budapest in May 1944—at the age of nineteen and at a time when deportations from the Hungarian countryside were at their peak—was identical to the Auschwitz Report of Vrba and Wetzler (1).”

Next in this remarkable tale, Klein decided to try to find Vrba, to “tell him of what enormous help his report had been to me. If I had not known what was awaiting me at the other end of the train trip, I would never have dared to risk an escape. It was not difficult to find Vrba, for it turned out that we were scientific colleagues. He is a professor of neuropharmacology in Vancouver, and I am now (in the Spring of 1987) sitting in a comfortable armchair in the faculty club at a Canadian university, talking with someone who, at first glance, seems quite ordinary. He impresses me as being relaxed and jovial. By now I have also read his book (Escape from Auschwitz, 1964), and I am aware that he has survived more death sentences than anyone else I have ever met (1).”

Vrba (1924–2006), was indeed a professor of pharmacology at the University of British Columbia; a position he held from 1976 until the early 1990s. Note that he and Wetzler were the first prisoners ever to escape from Auschwitz. Vrba’s real name was Walter Rosenberg. Rudolf Vrba was the nom de guerre he used after joining the resistance in his native Czechoslovakia. Afterwards, he made the change legal.

The horrors of the holocaust remained an obsession for Klein, although he was uncertain as to why that was so. “Was it to honor my murdered family, my murdered classmates? Or was it rather to steel myself against the darkest side of our human heritage?” In any case, Auschwitz and the holocaust were the main topics of conversation when Klein met with Vrba.

Vrba took Budapest’s Jewish Council to task for not widely broadcasting the warnings in the Auschwitz Report. He, and others, have alleged that Dr. Kastner, a well-known Zionist leader in Budapest, decided to keep the Report secret, in return for a promise from the Germans to allow sixteen-hundred people, as selected by Kastner, to safely emigrate from Hungary. Klein retorted that he knew Kastner from his work for the Jewish Council, and considered him to be a hero, because he had rescued many, while others tried to rescue only themselves or their own families. [In 1957, Kastner was murdered in Israel by a young man whose family was exterminated by the Nazis. Kastner remains a controversial figure to this day.]

Klein and Vrba next discussed whether dissemination of the Auschwitz Report might have caused Budapest’s Jews to revolt against the Nazi program of annihilation.  Klein argued that of the dozen or so people that he warned, no one believed him. Vrba countered, “You were a mere boy. Why would anyone believe what you were saying? The Jews would certainly have believed their responsible leaders (1).” Nonetheless, Vrba conceded that even the prisoners at Auschwitz were in denial of what they could see with their own eyes: “…prisoners, who knew full well that no one ever returned from the gas chambers, repressed such knowledge as they themselves lined up for execution in front of the chamber doors.”

Klein asked Vrba how he is able to live and function in Vancouver, a pleasant and friendly place, where no one has the slightest concept of what he endured: “…you must go back constantly to those days. You are called in as a witness at trials of old Nazis or their followers, people who claim that the holocaust never happened. You try to describe something that cannot be described in any human language, you try to explain the incomprehensible, you want people to listen to something they do not want to hear (1).” Vrba, in fact, never did reveal his Auschwitz experience to his colleagues. Vrba explained: “What would have been the use? No one who has not experienced it can understand.” Their conversation went on for almost ten hours. Afterwards, they parted like old friends, despite any differences in their views.

In the Fall of that year, Klein was reunited with Vrba in Paris, together with another newfound friend, German scientist Benno Muller-Hill. In 1966, Muller-Hill was a graduate student in Walter Gilbert’s Harvard laboratory, when he purified the lac repressor; the first genetic control protein to be isolated. Muller-Hill then began a second career lecturing and writing about the role of Nazi doctors and scientists in the holocaust. Klein met Müller-Hill for the first time at a meeting at the Institute for Genetics in Cologne, and the two immediately developed a close friendship.

Muller-Hill was in Paris to visit colleagues at the Pasteur Institute, as well as to meet Vrba. Klein was visiting Paris after attending a scientific meeting in Lyon. Vrba was in Paris at the invitation from the French radio service to refute claims of the ultra-right French leader, Jean Marie Le Pen, that the Nazi gas chambers never existed, and that if the Nazis indeed had any intent to annihilate the Jews, it was merely one of many episodes of the war. [Marine Le Pen, currently a leader of France’s ultra-right National Front, and a candidate for the presidency of France, is Jean Marie’s daughter. She was recently taken to task for denying that French officials and police were complicit in the Nazi roundup of more than 13,000 French Jews in July 1942 (they were later deported to Auschwitz). Le Pen also calls for the deportation of all immigrants from France; a stance that mainly targets Muslims.]

Klein and his two companions ambled about Paris on a beautiful Fall afternoon. They strolled around the Luxembourg Gardens, then continued along the banks of the Seine, turned toward the Latin Quarter, and then stood before the façade of Notre Dame. Yet their minds were elsewhere. “Vrba suggested that we visit the holocaust memorial behind Notre Dame…That walk of only a few minutes took us from the noisy tourist crowd to the silence of the museum’s rooms, where you feel alone and isolated among the symbolic chains and barbed wire. A faint glow of sunlight came in through the narrow openings in the wall. We were surrounded by the voices of the victims…We were all completely speechless. Even Vrba’s macabre sense of humor and his sharp sarcasm had fallen silent for the moment (1).”

After they exited from the memorial, they sat down in a small bistro, where Klein asked his two companions whether German scientists and doctors were actual architects of the holocaust or, instead, merely passive followers. “Benno had concluded from his exhaustive documentation that, contrary to what many wanted so desperately to believe, the ‘euthanasia programs’…and the horrible human experiments… could not be ascribed to a small minority of madmen, opportunists, or charlatans. On the contrary, they had been carried out by quite ordinary and in some instances, eminent physicians and scientists… He (Verba) thought … that would not explain why so many apparently ordinary people took part in the murders without showing any signs of remorse, or how the annihilation program could have been carried out with such efficiency… The discussions between Benno and Vrba continued for several hours (1).”

The day became even more notable later, since Klein had arranged for the threesome to have dinner that evening with Francois Jacob. After a glass of sherry in Jacob’s Latin Quarter apartment, the foursome went to a small restaurant around the corner.

Francois Jacob, and fellow Pasteur Institute scientist Jacques Monod, were awarded Nobel Prizes for their work together on the regulation of lactose metabolism in E. coli (2). More apropos the current episode, Jacob and Monod each received France’s highest military honors for his service during the Second World War—Jacob for his heroism serving with the Free French forces, and Monod for his heroism in the Resistance (2). Yet Jacob’s harrowing escape from Nazi-occupied France at 19-years in age, and his wartime exploits as one of Charles De Gaul’s most highly decorated volunteers, were barely known to his three dinner companions.

At first, Klein was somewhat worried that his friends might not like each other. Jacob often found conversation to be difficult; partly because the thousands of pieces of shrapnel that he carried in his body from the war, made it hard for him to sit comfortably. [Jacob’s wartime wounds prematurely ended his surgical career, and led him to turn to a career in science (2).] But, the get-together didn’t go badly at all.

Conversation eventually turned to the issue of holocaust deniers, as well as to those who would put the past completely behind them. As they talked, the incongruity of the scene suddenly struck Klein. They were sitting in a “first-class Parisian restaurant, surrounded by elegant people, having a very nice dinner in the best French tradition.” “…why did the three of us, with Jacob listening, choose to spend that beautiful Saturday in Paris compulsively focusing our attention on the black birds? We were all citizens of free countries, living well in peaceful times. Were we haunted by feelings of guilt toward the dead? Were we afraid that the whole experience would recur if we let go? We knew that the wide and relentless river of history is rarely influenced by knowledge of the past. In no more than one or two generations, archives of extreme horror turn into scraps of faded paper, with no more influence than dried leaves. I suddenly felt that we were like a traveler with a fear of flying, forcing himself to stay awake and keep his seatbelt buckled during the entire flight, obsessed with the idea that the plane would surely crash if he were to fall asleep. But perhaps we had other motives. Perhaps we wanted to feel a solidarity with each other by selecting a more or less taboo subject for our conversation, one avoided by most others. Or did we try to perform a kind of autopsy, using our brains to understand what human minds are capable of at their worst? Have we appointed our brains to serve as the pathologist and the cadaver at the same time?”

The above recounts only a small sampling of Klein’s conversations with Vrba, Muller-Hill, and Jacob, during their day together in Paris. For more, see reference 1.

In January 1945, 19-year-old George Klein emerged from the Budapest cellar where had been hiding during the last weeks of the German occupation. He gazed on the dead soldiers, civilians, and horses that were frozen in the snow, and was struck by the thought that he had survived, despite the likelihood that he would have ended his 19 years in a Nazi gas chamber or a slave labor camp. However, with the city now in Russian hands, George faced a new threat to his freedom; the Russian patrols that were exporting young Hungarians to labor camps in Russia.

Mindful of the danger on the streets, George was yet eager to begin his medical studies. So, he cautiously dodged the Russian patrols as he made his way to Budapest’s medical school, only to find war-torn deserted buildings and dead soldiers there.

Undeterred by the situation in Budapest, George and a friend set out to Szeged, with the hope of attending the medical school there. The journey of 160 miles took the pair five days, by way of a variety of vehicles, including a Russian military truck. In any case, they were admitted to the Szeged university on the same day that they arrived. And while the school was a shadow of its former self, with all the professors having fled to the West, to George, it was a “previously forbidden paradise (3).”

George spent two years in Szeged, and then returned to Budapest when the University reopened there. Back in Budapest, George fell “desperately” in love with Eva Fisher, a fellow medical student. [George describes their whirlwind romance in reference 3.] George now faced a dilemma. Before he met Eva, he finalized plans to visit Stockholm (under the sponsorship of the Jewish Student Club there). But going to Stockholm would mean leaving Eva behind, under conditions in which travel back into Hungary could be risky. Nonetheless, George went to Stockholm, with Eva believing she would never see him again. Yet the trip would be a defining experience for George and, eventually, would be important for Eva too. In Stockholm, George would learn of, and be riveted by the research of renowned cell biologist Torbjörn Caspersson, at Stockholm’s Karolinska Institute.

Caspersson’s research so enthralled George that he diligently pressed Caspersson for a junior research assistantship in his laboratory. But once George had been accepted by Caspersson, he viewed his situation with a “mixture of ecstatic happiness and enormous anxiety.” “I knew virtually nothing…I was halfway through my medical studies…I was desperately in love with a girl whom I had only known during a summer vacation of eight days and who was on the other side of an increasingly forbidding political barrier (3).”

Despite these misgivings, George knew that his future lay in Sweden, rather than Hungary. He had been accepted into Caspersson’s laboratory, and Hungary was falling increasingly under totalitarian Soviet domination. But Eva was still in communist Hungary. So, George risked returning there with one goal; to marry Eva, and then to leave Hungary for good. “The reunion with Eva confirmed what we both already knew: we wanted to live and work together (3).”

But George and Eva didn’t have the necessary documents to get married, nor did Eva have a passport to leave Hungry. Moreover, communist bureaucrats made it increasingly difficult to obtain these documents. In some instances, up to six weeks might be needed. However, George and Eva were daring and resourceful. When told by a police officer that it would take at least three weeks to obtain a marriage license, George suddenly acted on impulse: “I had always heard others tell of such things but I myself had neither seen nor done it. I pulled a fairly modest bill out of my pocket and put it in the policeman’s hand. ‘Pardon me, how much time was it you said?’ ‘I’ll go get it at once,’ he answered (3).”

With similar persistence and ingenuity, George and Eva obtained all their necessary documents, and they were married that very day! One document, a certificate asserting that neither George nor Eva had a venereal disease, would normally require a three-week lab test. But they beseeched an older colleague, now a doctor at a children’s hospital, to write the certificate for them. Their colleague did so, on his Children’s Hospital stationary. George and Eva then went to the prefecture to be married, only to find a disagreeable marriage official, who was determined to leave work for the day. But, when the official leafed through their papers, and saw the venereal disease certificate written on Children’s Hospital stationary: “He laughed until tears ran down his cheeks. This was the funniest thing he had seen during his whole time in service.” He then gladly married the couple.

As the Iron Curtain descended about Hungary, George and Eva left for Sweden, where they would now continue their medical studies. What’s more, Eva joined George in Caspersson’s laboratory at the Karolinska Institute. The couple would work together at the Karolinska until George’s death at the age of 91. [Eva was born to Jewish parents in Budapest in 1925. In 1944 and 1945, she and several members of her family hid from the Nazis at the Histology Institute of the University of Budapest. Encouraged by Caspersson, Eva had an independent research career, while also collaborating with George. She is best known for discovering natural killer cells, and for generating the Burkitt’s lymphoma cell lines, which she and George studied together (see below).]

George and Eva, at the Karolinska Institute, 1979

We conclude with a brief review of some of George Klein’s contributions to virology and to cancer research.

Tumor immunology: In 1960, George and Eva used methylcholanthrene to induce tumors in mice. Next, they surgically removed the tumors, killed them with irradiation, and inoculated them back into genetically compatible mice. Next, they challenged these mice with cells from a variety of different tumors, and showed that the immune systems of the inoculated mice rejected only those cancer cells that came from the original tumor. Thus, there are tumor-specific antigens that can be recognized by the immune system. See Aside 1.

[Aside 1: Importantly, the tumor resistance seen in these experiments did not arise spontaneously in the original tumor-bearing animals. Instead, it developed in the test mice, in response to sensitization with killed tumor cells. Thus, these experiments per se do not point towards an immune mechanism of tumor surveillance. Nonetheless, harnessing such a mechanism is currently a promising means of cancer therapy, and was a major theme in Klein’s thinking.]

The following year, Klein’s group showed that polyoma virus-induced tumors share a common antigen. Importantly, polyoma virus-induced tumors, and polyoma virus-transformed cells, were rejected irrespective of whether they released virus. Thus, antiviral immunity as such was neither necessary nor sufficient for tumor rejection. This was the first demonstration that tumors caused by a virus might share a common antigen. The Kleins, and others, later found similar “group-specific” transplantation antigens on other virus-induced tumors, including retrovirus-induced lymphomas.

Burkitt’s lymphoma: “Sometime in the mid-1960s, Eva suggested that we should use our experi­ence on virus-induced murine lymphomas to examine a human lymphoma with a presumptive viral etiology. Could we detect group specific antibody responses that might be helpful in tracing a virus? Burkitt’s lymphoma (BL) was the obvious choice (3).” [Burkitt’s lymphoma, originally described by Dennis Burkitt in 1958, is a malignant B-cell lymphoma that is most prevalent in tropical Africa and New Guinea. It is the most common childhood cancer in equatorial Africa. Burkitt first proposed that the lymphoma might have a viral etiology, since its geographic distribution is like that of yellow fever, which is caused by a flavivirus. In 1964, Tony Epstein and Yvonne Barr, by means of electron microscopy, discovered a virus in cells which they cultured from BL tissue, thereby giving credence to Burkitt’s premise.]

Klein’s group identified a membrane antigen (MA) that was expressed in some BL-derived cell cultures. Werner and Gertrude Henle had previously discovered that the MA antigen is a structural protein from a newly discovered herpesvirus—the virus that Epstein and Barr first saw in 1964. Klein decided to call that virus the Epstein Barr virus (EBV). The MA antigen is now known to be one of the EBV envelope glycoproteins. Klein and collaborators later identified complement receptor type 2 (CR2), also known as the complement C3d receptor, as the cell surface attachment protein for the viral MA glycoprotein. CR2 receptors on B cells play a role in enabling the complement system to activate B cells.

By 1970, Klein’s group, in collaboration with Harald zur Hausen, found that the subset of BL-derived cell lines that express MA are, in fact, those that produce EBV. However, more than 90% of the BL cell lines, and all nasopharyngeal carcinomas, were found to contain multiple EBV genomes per cell, irrespective of whether they produced virus. Thus, only a subset of BL and nasopharyngeal carcinoma cells that harbor EBV genomes, actually produce the virus. During this time, the Henles discovered that EBV is the cause of infectious mononucleosis, and that EBV could immortalize normal B cells in culture.

Oncogene activation by chromosomal translocation: A sero-epidemiological study, begun in Uganda in 1971 by Geser and de-The, showed that children with a high EBV load are more likely to develop BL than are children with a low EBV load. Thus, the presence of EBV genomes in a B cell increases the likelihood of it turning into a BL. “But this is still not a satisfactory explanation; some essential element is obviously missing (3).”

What then is the missing event that gives rise to BL? A 1972 study by Manolov and Manolova, Bulgarian scientists working with the Kleins, found that a particular chromosomal marker, 14q+, was present in about 80% of BL tumors. After the Manolovs returned to Bulgaria, the Kleins, in collaboration with Lore Zech, used the chromosomal banding technique recently developed by Caspersson and Zech to examine the BL-cell chromosomes more precisely. They showed that the 14q+ marker was derived from chromosome 8, which broke at the same site (8q24) and underwent a reciprocal translocation with the short arm of either chromosome 2 or chromosome 22. All BLs carried one of the translocations.

Meanwhile, another research group found that carcinogen-induced mouse plasmacytomas are associated with an almost homologous chromosomal translocation. Thus, a common mechanism seemed to underlie two distinct types of tumors, in two distinct species. In each instance, a putative oncogene was translocated to an immunoglobulin locus, which might then have caused the oncogene to be constitutive expressed. A somewhat similar mechanism was reported earlier for the induction of bursal lymphomas in chickens by the avian leukosis virus (ALV) . In that instance, the cellular c-myc gene came under the control of the ALV provirus promotor. What’s more, Michael Cole’s group identified the transposed gene in BL, and in the mouse plasmacytomas, as c-myc. It is not yet clear how EBV infection promotes the chromosomal translocation.

Tumor suppressor genes: In the early 1970s, Klein, and collaborator Henry Harris, played a pioneering role in developing the concept of tumor suppressor genes. They found that when highly malignant mouse cells are fused with normal mouse cells, the hybrid cells are non-malignant when inoculated into genetically compatible mice. That is, tumorgenicity is suppressed by fusion with normal cells. However, tumorgenicity reappears after some apparently important chromosomes, contributed by the normal cell, are lost from the hybrid cells.

References:

  1. George Klein. Pieta, MIT Press, 1992.
  1. The Converging Lives of Jacques Monod, Francois Jacob, Andre Lwoff, and Albert Camus in Wartime France, Posted on the blog March 27, 2017.
  1. George Klein and Eva Klein. 1989. How One Thing Led to Another, Annual Review of Immunology, 7:1-33.

The Converging Lives of Jacques Monod, Francois Jacob, Andre Lwoff, and Albert Camus in Wartime France

In the 1950s and 1960s, two French biologists at the Pasteur Institute, Francois Jacob and Jacques Monod, explained how genes are regulated in bacteria. Their studies of the “lac operon” of E. coli indeed opened up the field of gene regulation, and were a key development in the new science of molecular biology. Their experimental findings also implied the existence of an unstable intermediate between genes and protein synthesis, which eventually led to Jacob’s discovery, in collaboration with Sydney Brenner and Matt Meselson, of messenger RNA (1).

Jacob and Monod shared in the 1965 Nobel Prize for physiology or medicine for their breakthrough studies on gene regulation. Fellow Pasteur Institute scientist, Andre Lwoff, received a share of the award for his pioneering studies on the nature of lysogeny (i.e., how a bacteriophage’s genome can be incorporated into the genome of a host bacteria, and remain latent until being activated by an inducing factor).

In 2013, evolutionary biologist Sean B. Carroll published a book—Brave Genius: A Scientist, a Philosopher, and their Daring Adventures from the French Resistance to the Nobel Prize—that relates how wartime circumstances brought together Jacques Monod and his scientific colleagues Francois Jacob and Andre Lwoff (2). But while much of that story is already known (3), Carroll also tells us of the little known, but remarkable coming together of Monod and philosopher/writer Albert Camus, one of the intellectual giants of the 20th century. Coming from very different intellectual backgrounds, Monod and Camus forged a deep friendship, united in their opposition to tyranny and oppression. Carroll’s book was the inspiration for this post.

When Albert Camus learned that he had won the Nobel Prize for Literature in October 1957, he wrote to a few well-wishers, including an old friend in Paris:

My dear Monod,

      I have put aside for a while the noise of these recent times in order to thank you from the bottom of my heart for your warm letter. The unexpected prize has left me with more doubt than certainty. At least I have friendship to help me face it. I, who feel solidarity with many men, feel friendship with only a few. You are one of these, my dear Monod, with a constancy and sincerity that I must tell you at least once. Our work, our busy lives separate us, but we are reunited again, in one same adventure. That does not prevent us to reunite, from time to time, at least for a drink of friendship! See you soon and fraternally yours.

                                                                                                                                             Albert Camus

Camus appears somewhat downcast in his note to Monod. At 43-years-in-age, he was the second youngest writer ever to receive the Nobel Prize for literature (Rudyard Kipling at 42 was the youngest), and he was worried that the ballyhoo surrounding the award might distract him from his writing. And, he was concerned that the prize might stir up additional contempt from critics of his writing, as well as from his leftist colleagues who opposed his condemnation of Soviet communism.

But, why did philosopher/writer Camus—an intimate of some of the greatest writers and artists of the mid-twentieth century, including Sartre and Picasso—write to scientist Monod, and acknowledge the special importance he placed on their friendship? Likewise, why did he assert: “I have known one true genius, Jacques Monod.” And, what is the same adventure that Camus refers to?

A brief background to our tale is as follows. In March 1939, Hitler took control of Czechoslovakia. Next, on September 1, Germany invaded Poland. On September 2nd, Poland’s allies, Britain and France, issued an ultimatum to Germany: withdraw or face war. On September 3rd, the ultimatum expired, Britain and France declared war on Germany, and the Second World War was underway; sort of. Although Germany went on to conquer Poland in a mere eight days, several months passed without further action. Then, in May 1940, Nazi Germany invaded and overran France in just six weeks. Marshall Pétain surrendered to the Germans, the French Forces were disbanded, the pro-Nazi Vichy government was put in place under former prime minister Pierre Laval and the 84-year-old Pétain, and the Nazi occupation of the defeated French nation began.

A few months before the Nazis invaded France, thirty-year old Jacques Monod was a doctoral student in zoology at the Sorbonne. [A polymath, he also founded a Bach choral group, and was an accomplished cellist, and seriously considered a career in music (4).]  But as war with Germany loomed, Monod enlisted in the army—in the communication engineers—where he thought he might use his scientific talents if war were to break out. Consequently, Monod was serving on the front lines when the Germans invaded. France suffered the most colossal military disaster in its history, and Monod returned to his studies in Paris.

Life in France grew progressively harsher under the Nazis; beginning with subjugation, and followed by deportations, enslavement, and mass murder. Early on, Monod joined one of the first units of the French Resistance; a group of ethnologists and anthropologists at the Musée de l’Homme (Museum of Man).

One of Monod’s duties for the Musée de l’Homme group was to distribute its newspaper, at night. This seemingly simple task was extremely dangerous since capture could mean deportation to a concentration camp or execution. Monod, in fact, had several close escapes. On one occasion, the Gestapo raided his laboratory at the Sorbonne. Fortunately, since they were fearful of the viruses and radioactive isotopes in the lab, they didn’t search it as thoroughly as they might have. Otherwise, they might have found sensitive documents that Monod would hide inside the leg of a mounted giraffe outside his office. In any case, the Germans soon routed the short-lived Musée de l’Homme group

Monod’s wife, Odette, was the granddaughter of Zadoc Kahn, the former chief rabbi of France. Since the Vichy government soon began enacting Nazi policies, including anti-Jewish laws, and because of homegrown French anti-Semitism, Odette sought refuge for herself, and for her and Jacque’s twin sons (born in August 1939, four weeks before the war broke out), under assumed names, in a village outside of Paris. Meanwhile, Jacques had to register with the Vichy authorities as the spouse of a Jewish person.

With Odette and the children concealed, Monod joined the most militant unit in the Resistance; the Communist-led Franc-Tireurs (Free Shooters) group. Monod was not then a Communist Party member. Nonetheless, he joined the Franc-Tireurs since they actually were fighting the Germans—assassinating German officers in the streets and carrying out sabotage. One of his missions for the Franc-Tireurs took him to Geneva—through the Alps to avoid arrest—to request money for arms from the United States Office of Strategic Services; the precursor of the present Central Intelligence Agency.

By this time, Monod had gone completely underground. He wore a disguise during the day, slept in safe houses at night, and stayed away from his laboratory at the Sorbonne. But then, Andre Lwoff, the head of microbial physiology at the Pasteur Institute, offered Monod a refuge and a place to work in his laboratory at the Pasteur Institute. Monod then led a double-life. By day, as Monod, he worked on his experiments at the Pasteur Institute. At night, he carried out his duties for the Franc-Tireurs, as “Marchal” (from a character in a novel by Stendhal), and as  commander “Malivert.” [Lwoff too had been active in the Resistance, gathering intelligence for the Allies, while also hiding downed American airmen in his apartment.]

Jacques Monod’s identity card for the French Forces of the Interior (FFI), in his nom de guerre ‘Malivert.’

Monod was resolutely committed to the Resistance, while also maintaining a productive research program. At the Pasteur Institute, he and his student, Alice Audureau, made key discoveries that would lead to the later breakthroughs he would make with Jacob. [For instance, Monod and Audureau discovered mutations in E. coli genes that caused the induction of lactose metabolism; a finding that would have important implications concerning gene action and regulation.] Moreover, he was devoted to Odette and their twin sons, and managed to make frequent clandestine visits to see them.

Monod took on increasing responsibilities in the Franc-Tireurs, as more members of the group were discovered and executed by the Germans. In fact, by the time of the allied invasion of Normandy in June 1944, Monod, had become chief of staff of the operations bureau for the National Resistance Organization; a position from which his three predecessors had disappeared (4). As such, Monod prepared battle plans for the allied surge to Paris. He also arranged parachute drops of weapons, railroad bombings, and mail interceptions.

Interestingly, Monod also recruited to the Resistance renowned French chemist, John Frédéric Joliot-Curie (Aside 1), who devised a unique recipe for Molotov cocktails, which were the Resistance’s principal weapon against German tanks. In addition, Monod organized the general strike that facilitated the liberation of Paris. Then, after the liberation of Paris, he became an officer in the Free French Forces, and a member of General de Lattre de Tassigny’s general staff.

[Aside 1: John Frederick Joliet was working as an assistant to Marie Curie, when he married Marie’s daughter, Irene. Afterwards, both John Frederick and Irene changed their surnames to Joliot-Curie. In 1935, the couple was awarded the Nobel Prize in Chemistry for their seminal research on radioactivity. John Frederick then worked at the Collège de France on controlled chain reactions. His work on that was cited by Albert Einstein in his famous 1939 letter to President Franklin Roosevelt, warning Roosevelt of the possibility of a nuclear weapon: “In the course of the last four months it has been made probable through the work of Joliot in France as well as Fermi and Szilard in America—that it may be possible to set up a nuclear chain reaction in a large mass of uranium, by which vast amounts of power and large quantities of new radium-like elements would be generated. Now it appears almost certain that this could be achieved in the immediate future…This new phenomenon would also lead to the construction of bombs…” The Nazi invasion ended Joliet-Curie’s nuclear research. Nevertheless, he managed to smuggle his research notes out of France to England.]

Francois Jacob, a Jewish, nineteen-year old 2nd-year medical student, was planning on a career in surgery when the German occupation of France began in the Spring of 1940. Resolved to carry on the fight against Hitler, Jacob left medical school and boarded one of the last boats for England. In London, he was one of the first of the French to join Charles de Gaulle’s Free French Forces. He wanted to enroll in a combat unit, but, despite his incomplete medical training, he was commissioned as a medical doctor, and then served as a medical officer in North Africa. His surgical career was prematurely cut short in August 1944, when he was severely wounded at Normandy; by a bomb dropped from a German Stuka dive bomber. At the time, he was tending to a dying officer.

For their wartime service, Jacob and Monod were each awarded France’s highest honors for valor. Jacob was awarded the Cross of Liberation, as well as the Légion d’Honneur and the Croix de Guerre. Monod likewise received the Légion d’Honneur and the Croix de Guerre, as well as the American Bronze Star.

Unable to practice surgery after the war because of his wartime wounds, Jacob eventually turned to a career in science. He was accepted at the Pasteur Institute, where he beseeched Lwoff (Monod’s host at the Pasteur Institute) to serve as his mentor. Lwoff rebuffed Jacob several times, but finally agreed to take the young doctor under his wing. Then, in the cramped quarters of Lwoff’s laboratory at the Pasteur, Jacob and Lwoff’s student, Elie Wollman, began a fruitful collaboration that produced key insights into bacterial conjugation and the regulation of lysogeny (Aside 2). After that, Jacob and Monod forged their extraordinary collaboration that would lead to their Nobel Prizes. Note that Jacob’s earlier work with Wollman, on lysogenic induction, would provide the underpinning for his later work on gene regulation with Monod (3).

[Aside 2: Elie Wollman, born in 1917, was Jewish. In 1940, he escaped from the Nazis in Paris and then worked underground in the Resistance as a physician. His parents, Eugene and Elizabeth Wollman, were Pasteur Institute scientists who were seized by the Nazis in 1943 and sent to Auschwitz. They were never heard from again (3).]

In December of 1939, our other main protagonist, twenty-six-year-old Albert Camus, was an unknown, aspiring writer, working as a reporter and editor for a newly founded left-wing newspaper, Alger Republican, in his native Algeria; which was then under French control. Camus was completely opposed to the war, which he saw as “another unnecessary, avoidable, disastrous, absurd chapter of history that would consume the lives of those who did not make it or wish for it.” His antiwar editorials in the Alger Republican outraged French government officials who were calling for unity against Germany. The government finally shut down the newspaper, leaving Camus unemployed. So, Camus returned to France, where the prospects for employment were now better because wartime mobilizations had left many businesses shorthanded. See Aside 3.

[Aside 3: Camus started writing The Stranger while in Algeria, basing it on people and places he knew there. His purpose in The Stranger was to express how one might react to his philosophical notion of the “absurd”—the disconnect between our desire for a rational existence, and the actual world, which appears confused and irrational—in the form of a novel. Meursault, the narrator, and principle character in The Stranger, shows no grief over his mother’s death, no remorse over having committed an unintended murder, and no belief or interest in god. Even while Meursault was awaiting the guillotine, he was reconciled to “the tender indifference of the world.” Meursault’s honesty in describing his feelings makes him a ‘stranger’ in the setting of the novel, and seals his fate.]

Camus was not called up for military service when he returned to France, because he had contracted tuberculosis in Algeria, when he was 17 (Aside 4). Nonetheless, he twice attempted to enlist—the second time when the French Army was on the verge of surrender to the Nazis—to express his solidarity with those who were being drafted. In any case, the military rejected him each time because of his tuberculosis. So, he managed to get a job in Paris as a layout designer for the newspaper Paris-Soir.

[Aside 4: In the pre-antibiotic era, tuberculosis was often fatal, and the 17-year-old Camus indeed had a close brush with death. That experience had a profound effect on the “precocious philosopher,” who made notes on the question of “how, in the light of the certainty of death, one should live life.”]

Parisians began fleeing from their city when the German invasion began in May of 1940. Then, in June, as the Germans were on the verge of entering Paris, the stream of refugees became a flood, with about 70 percent of the city’s metropolitan population of nearly five million eventually taking flight from the city. All Parisian newspapers stopped publishing. However, Paris-Soir hoped to resume its operations in the south, with a reduced staff. Thus, Camus joined the stream of refugees, driving an automobile (almost all the paper’s regular drivers had been drafted), with a Paris-Soir executive as his passenger. After Camus and his passenger were well on their way, Camus suddenly realized that in the rush to vacate from Paris, he may have left his manuscript for The Stranger behind in his room. “He jumped out of the car and threw open the trunk, and was relieved to find in his valise the complete text of The Stranger.” See Aside 5.

[Aside 5: In 1885, Joseph Meister, at nine-years-of-age, was the first recipient of Louis Pasteur’s rabies vaccine and, as an adult, was caretaker of the Pasteur Institute; a position that he still held at the start of the Nazi occupation in 1940.  In despair over the fall of France, and wrongly believing that German bombs killed his family after he sent them away, he went to his apartment, closed the windows, and turned on the gas in his stove (5, 6).]

Camus went with Paris-Soir to Clermont-Ferrand. There, the paper began to publish again, using printing facilities made available by Pierre Laval, the former premier, and now architect of the Petain Vichy government. But with the paper now under Laval’s control, it began publishing anti-Semitic articles, and other articles in support of the Vichy government. Camus did not write any of these items. In any case, he was let go by Paris-Soir after the draftees of the 1940s were discharged and could return to work. Camus then went back to Algeria, where he completed The Stranger.

In 1942, with The Stranger about to be published in France, Camus suffered a nearly fatal relapse of his tuberculosis. He wanted to return to France for treatment in the Massif Central mountain range, but several months would pass before Algerian authorities gave him permission to do so. Then, upon returning to Paris, he would have a purpose that would totally engage him.

One night, under an assumed name (because of the need for secrecy in the Resistance), Camus stole into the clandestine headquarters of Combat (the journalistic arm and voice of the French Resistance), to implore the staff to take him on since he “had already done a little journalism” and would be happy to help in any way. Like Monod, Camus then led a double-life, carrying out his duties at Combat, as “Bauchard.” At first, he helped to select and edit articles, and prepare the paper’s layout. Then, in 1943 he became the paper’s editor, and wrote stirring editorials, exhorting Frenchmen to act against the German occupiers. By the time The Stranger was published in 1942, his recognition as Camus led to his acceptance into the literary and artistic circle that included Sartre, Simone de Beauvier, and Picasso.

Albert Camus’s false identity card, in the name of Albert Mathé, writer. All of the information on the card — birthdate, place, parents — is false.

Camus was suffering from recurrent bouts of tuberculosis all the while that he was carrying out his work at Combat. Nonetheless, as Camus, he also published his essay, The Myth of Sisyphus, which, like The Stranger, contemplates the experience of the Absurd (see Aside 3, above). And he also wrote The Plague, which depicts a city’s response to an outbreak of bubonic plague; perhaps a metaphor for the Nazi occupation. Remarkably, no one at Combat had an inkling that the man who at first had been editing and arranging pages for them as Bauchard was in fact the now renowned Camus. See Aside 6.

[Aside 6: Among laypeople, Jacques Monod is perhaps best known for his “popular” book, Chance and Necessity, published in 1970, and a bestseller in its day. Monod’s Chance and Necessity, and Camus’ The Myth of Sisyphus, are each relevant here because they point up how Camus influenced Monod’s view of the meaning of life. While Camus took a philosophical approach to that issue, Monod’s assessment was also informed by his knowledge of life’s fundamental molecular mechanisms. With the 1953 discovery by Watson and Crick of the molecular structure of DNA, it was apparent how accidental, random, unpredictable mutations in the sequence of bases in DNA were the source of all biological diversity. Thus, Monod knew that all living forms, including humans, are the products of chance genetic mutations and circumstances: “Man at last knows that he is alone in the unfeeling immensity of the universe, out of which he emerged only by chance. Neither his destiny nor his duty have been written down. The kingdom above or the darkness below: it is for him to choose.” [Monod’s title, Chance and Necessity, is from Democritus’ dictum “Everything in the universe is the fruit of chance of chance and necessity.”]  

That we live in a world that is indifferent to our hopes and suffering was the reason for Monod to inquire into the meaning of life, which, for Camus, was “the most urgent of questions.” Camus was often branded an existentialist, but unlike many contemporary existentialist thinkers, Camus vehemently rejected nihilism. In The Myth of Sisyphus, he wrote that Sisyphus gave his life meaning by choosing to believe that he remained the master of his own fate, even though he was condemned to rolling his rock uphill each day, only to have it roll back down.

On the opening page of Chance and Necessity, Monod includes a lengthy quotation from the closing paragraphs of The Myth of Sisyphus. “The struggle itself towards the heights is enough to fill a man’s heart…One must imagine Sisyphus happy.” Camus is advocating that we oppose the certainty of death in an uncaring Universe by living life to the fullest. For Monod, life is like Sisyphus, pushing its rock uphill. The end might be bleak, but “the struggle towards the heights is enough to fill a man’s heart.”]

By 1944, the liberation of Paris was imminent, Combat went from a monthly publication to a daily one, and the paper chanced to circulate in the open. Camus was still writing his editorials anonymously. And when his identity was finally revealed, his inspiring, eloquent words resulted in his widespread public acclaim.

Monod and Camus were very likely aware of each other at this point in our saga, but they had not yet met. Their meeting would happen after the liberation of France, and it would be in response to a new totalitarian threat; from the Soviet Union. It transpired as follows.

In 1948, Monod was working full-time on his research at the Pasteur Institute, when events in the Soviet Union moved him to write a stirring editorial that appeared on the front page of Combat. [Camus had left Combat the previous year, after it became a commercial paper.] Monod’s piece was in response to a pseudoscientific doctrine advanced by Stalin’s head of Soviet agriculture, Trofim Lysenko, which asserted that organisms could swiftly change their genetic endowment in response to a new environment. [Lysenko’s doctrine is reminiscent of discredited Lamarckian doctrine, also known as heritability of acquired characteristics—i.e., the premise that if an organism changes to adapt to an environment, it can pass on those changes to its offspring.] Lysenko based his doctrine on his purported discovery of a means to enable winter wheat to be sown in the spring.

Stalin embraced Lysenkoism—during an acute grain shortage in Russia—since it was in accord with his ideology to create the New Soviet Man. Stalin also banned all dissent against Lysenko’s doctrine. Consequently, traditional Russian geneticists were exiled or murdered, Mendelian genetics was no longer practiced in the Soviet Union, and Soviet agriculture suffered severely.

Monod was roused to write his editorial after French Communist newspapers began to widely disseminate Lysenko’s doctrine in France. One Party newspaper proclaimed Lysenko’s discovery “A Great Scientific Event,” and further asserted that the notion of evolution by natural selection was a racist form of thinking, in harmony with Nazi doctrine (7). Another Party newspaper condemned Mendelian genetics for being “bourgeois, metaphysical and reactionary,” while claiming that it must be false because it is reactionary; having been invented by an Austrian monk. In Contrast, Lysenkoism is true because it is progressive and proletarian.

A Party member’s position on Lysenko indeed had become a gauge of his commitment to Stalin’s Soviet cause. But for Monod, the Soviet embrace of Lysenko was “senseless, monstrous, unbelievable.” As expected, Monod’s article was strongly condemned by the powerful French Communist Party, which enjoyed broad support from both intellectuals and workers; many of whom saw the Soviet Union as a model for a French socialist state. In any case, the Party’s strong backlash inspired Monod to “make his life’s goal a crusade against anti-scientific, religious metaphysics, whether it be from Church or State.” Importantly, a separate consequence of the Lysenko affair was that it influenced François Jacob to focus his research in the field of genetics. See Aside 7.

[Aside 7: Ironically, the observation that Jacob and Monod initially set out to explain looked remarkably like Lysenkoism. When E. coli are fed a solution of glucose and lactose, they grow rapidly until glucose—their preferred carbon source—is depleted. Only then, they turn to metabolizing lactose. But, in contrast to Lysenko’s doctrine, Jacob and Monod showed that when E. coli “adapts” to lactose, it does so without changing its genes. Instead, the genes encoding the enzymes that metabolize lactose lie dormant until lactose induces them, under conditions in which glucose is not available. That is, Jacob and Monod determined that lactose regulates lactose metabolism in the cell by acting as an inducer of genes that already exist in the cell; as opposed to lactose causing the cell to undergo a Lamarckian acquisition of a genetic characteristic. In so doing, Jacob and Monod created the now well-established paradigm of inducers, regulators, regulator genes, and operators.]

While Monod was crusading against Lysenkoism, Camus was having his own feud, in public, with Sartre, who had chastised him for his anti-Soviet stance. Camus had once been a Communist, in Algeria, mainly because he was troubled by the way in which the European French treated the native Algerians. However, he was never very sympathetic to the Marxist cause. Monod too had once been a member of the Communist Party; but only because it enabled him to have a voice in the running of the Resistance. In any case, Camus seized upon Monod’s condemnation of Lysenkoism in his feud with Sartre.

Our two main protagonists finally met when Camus co-founded the anti-Stalin, anti-totalitarian Groupes de Liaison Internationale. Monod attended one of the group’s meetings. There, he, and Camus, discovering that they shared much in common, forged their friendship. Carroll writes: “Camus, who so treasured the sense of solidarity that existed among the Resistance, had in Monod a new comrade who shared both the deep bond of that wartime experience and an unqualified opposition to a new common enemy.”

As noted, Monod’s views on the meaning of life owed much to Camus. Likewise, Camus learned from Monod. Camus not only used Monod’s case against Lysenko in his dispute with Sartre, but he also “borrowed” from Monod in The Rebel; in which Camus argued that revolution inevitably leads to tyranny. In any event, after Camus and Monod had separately fought the Nazis, they were now united against another oppressor—the totalitarian state run by Stalin. [Camus’ anti-Soviet stance cost him the friendships of many French intellectuals on the left. He and Sartre never spoke to each other again.]

Monod was also troubled by the situation of scientists working under Eastern European Soviet regimes. In 1959, he organized the escape into Austria of Hungarian biochemist Agnes Ullman (who participated in the failed Hungarian uprising of 1956), and her husband, also a scientist. Earlier, in 1958, Agnes Ullman managed to visit Monod at the Pasteur Institute, and confided to him that she and her husband wanted to defect from Hungary. Monod maintained contact with the Ullmans in Hungary, using coded messages, written in invisible ink, which turned blue when exposed to iodine. The Ullmans finally crossed into Austria, hidden underneath a bathtub, in a compartment of a pull-along camping trailer. See Aside 8.

[Aside 8. Agnes Ullmann, became Monod’s long-time close collaborator at the Pasteur Institute. Now retired, she was carrying out research at the Pasteur Institute as recently as 2012; 53 years after her rescue from Hungary. At the Institute, she collaborated with Monod on characterizing the lac operon promoter, on complementation between β-galactosidase subunits, and on the role of cAMP in overcoming the repressive effect of glucose (catabolite repression) on lactose metabolism in E. coli.]

There are numerous other instances in which Monod stepped forward to fight injustice and defend human rights. In 1952, he wrote a letter in Science that might have been “ripped from today’s headlines.” It protested the U.S. government’s rejection of visa requests for himself and other Europeans who had once been Communists. Monod also condemned the treatment of Jews in the Soviet Union, while continuing to speak out against Soviet totalitarianism in general. And, in 1965, shortly after Monod, Lwoff, and Jacob received word of their Nobel Prizes, they publicly appealed to the French government to allow the use of contraceptives, and the legalization of abortion. See Aside 9.

[Aside 9: Jacob too was devoted to the defense of human rights. He chaired a committee of the French Academy of Sciences that supported persecuted scientists living under totalitarian regimes, and he worked for the release of those who had been imprisoned for their political views. Moreover, he forcefully advocated for the public support of the biological and medical sciences. What’s more, Jacob also had a distinguished writing career that produced a series of acclaimed books, including The Logic of Life: A History of Heredity; Of Flies, Mice and Men; The Possible and the Actual, and his memoir, The Statue Within. In Joshua Lederberg’s review of the latter for The Scientist, he stated:As a work of literature, it evokes unmistakable overtones of Rousseau, Proust, and Sartre.” In Jon Beckwith’s view, all of Jacob’s books are “written in a fluid and elegant style” Others refer to the “clarity and grace” of Jacob’s writing. See reference 8 for more on Jacob.]

In 1966, Martin Luther King Jr. and Harry Belafonte visited France to raise funds for the Southern Christian Leadership Conference (SCLC). Remarkably, Monod was chosen to introduce King to a crowd of 5,000 people at Paris’ Palais des Sports. Belafonte was introduced by French singer and actor Yves Montand (9).

Coretta Scott King shaking hands with Jacques Monod, as Martin Luther King Jr. looks on. From l-r: French actress Simone Signoret, Harry Belafonte, French actor and singer Yves Montand. 29 March 1966, at a meeting of the “Movement for the Peace.”

The intellectual lives of Monod and Camus played out in entirely different areas. Yet the parallels were striking. Each, in his way, searched for meaning in life. Moreover, each put his life on the line to oppose ignorance, injustice, and totalitarianism. And, it is clear from their correspondences that they were dear to each other. Here is the note from Monod that elicited Camus’ response at the top of this post.

My dear Camus,

My emotion and my joy are profound. There were many times when I felt like thanking you for your friendship, for what you are, for what you managed to express with such purity and strength, and that I had likewise experienced. I wish that this dazzling honor would also appear to you, in some small part, as a token of friendship and of personal, intimate recognition. I would not dare coming to see you right now, but I embrace you fraternally.

Jacques Monod

This piece ends with a few personal thoughts. Jacques Monod was a Nobel Prize-winning scientist, a hero of the French Resistance, a rescuer of persecuted scientists from behind the Iron Curtain, and a leading voice against tyranny and oppression. And, he was also blessed with dashing good looks. I remember well the women among my fellow graduate students in the 1960s finding him to be very attractive. But, on a more serious note: Today, when political and religious blocs dismiss evidence-based science in favor of alternative ‘facts’ in order to advance their ideologies, and when they are tacitly aided by a press that all too often gives equal validity to all points of view, and while scientists seem to be groping for an effective response, one can hope that scientists with the courage, eloquence, and eminence of Jacques Monod and Francois Jacob might emerge to take up the cause of science and reason. Meanwhile, it is especially important for young scientists, and the public, to be aware of the examples set by these men. See Aside 10.

[Aside 10: The following is from a March 8, 2017 editorial in Nature.  “Last week, state legislators in Iowa introduced a bill that would require teachers in state public schools to include ‘opposing points of view or beliefs’ in lessons on topics including global warming, evolution and the origins of life… Since last month, Indiana, Idaho, Alabama, Texas, Oklahoma and Florida have all introduced and discussed similar tweaks to the way in which they want to educate their children… Although these proposed changes are typically presented by their supporters as giving teachers the chance to discuss genuine scientific controversies, in truth they are (very) thinly veiled attempts to pursue political and religious agendas that have no place in school science lessons — for whatever age. They seek to import the alternative facts and misleading rhetoric of the new federal government and to impose it on children who deserve much better from those elected to serve them.”]

References:

  1. A Most “Elegant” Experiment: Sydney Brenner, Francois Jacob, Mathew Meselson, and the Discovery of Messenger RNA , Posted on the blog October 6, 2016.
  2. Sean B. Carroll, Brave Genius: A Scientist, a Philosopher, and their Daring Adventures from the French Resistance to the Nobel Prize, Crown, 2013.
  3. Genealogies and a Selective History of Lysogeny: Featuring Friedrich Loeffler, Emile Roux, Andre Lwoff, Elie Wollman, and Francois Jacob, Posted on the blog January 28, 2015 (8).
  4. Agnes Ullmann, In Memoriam: Jacques Monod (1910–1976), Genome Biology and Evolution 3:1025-1033. 2011, DOI: https://doi.org/10.1093/gbe/evr024.
  5. Louis Pasteur: One Step Away from Discovering Viruses, Posted on the blog January 7, 2015.
  6. Dufour, H. D., and S. B. Carroll, History: Great myths die hard, Nature 502:32–33, 2013.
  7. John Marks, Jacques Monod, François Jacob, and the Lysenko Affair: Boundary Work, L’Esprit Créateur: Genetics and French Culture. 52:75-88.
  8. Beckwith, J., and M. Yaniv, Francois Jacob (1920-2013), Current Biology 23:R422-R425, 2013.
  9. Remembering Dr. Martin Luther King Jr., CBS Minnesota, January 19, 2013, 3:51 PM.

Confronting Science Denialism in the Trump Era

Our last post discussed President Trump’s advocacy of a link between vaccines and autism, while also noting that Trump has taken that stance despite the fact that the “link” has been well-debunked by several large-scale scientific studies (1). Also, our post made a case for biomedical scientists to speak out against Trump’s anti-vaccine pronouncements.

As Trump does when rejecting the findings of climate scientists, he similarly misrepresents and ignores the vast amount of scientific evidence that confirms the safety and effectiveness of vaccines. And this is happening while he asserts almost daily that any facts, which call his positions to account, are “fake.” Moreover, his millions of followers, who feast on his “alternative facts,” can pass them on to others with a click. See Aside 1.

[Aside 1: Trump surrogate, Scottie Nell Hughes, “explained” that everybody now had their own way of interpreting whether a fact was true or not.There’s no such thing, unfortunately, anymore as facts,” she declared. Thus, “a large part of the population” will pick and choose whatever “alternative facts” confirm their views (2).]

Many biomedical scientists now feel an urgent need to speak out against vaccine non-compliance. Yet others argue that scientists hurt the cause when they take political sides. Nonetheless, science is founded on honesty and rigor. And, if scientists do not speak out when their findings are distorted or ignored by politicians who put forward policies that harm the public, who else will? So, our concern here is to consider how we might effectively engage not only anti-vaxxers, but science denialists in general. It is important that we consider this, since we have not been especially effective in the past at curtailing science denialism (e.g., re evolution and human-caused global warming).

A key prerequisite for effective communication is that each party listen to, and acknowledge the others point of view. This may be difficult to accomplish with science denialists under any circumstance. But it is most difficult in public discussions, where a group of committed denialists is unlikely to allow the free and open discussion that is essential. Even if you should happen to get your points out, hard-core denialists in the audience will probably not consider them (see Asides 2 and 3). So, in front of a group, address your remarks to the skeptical and undecided members of your audience, rather than to the stanch denialists.

Your chance of influencing undecided or skeptical individuals is much greater in a one-on-one discussion. But whether before a group, or in a one-on-one discussion, your major asset and advantage is that the scientific consensus supports your position. Focus on the evidence.

[Aside 2: Hard-core denialists provide but one example of a more general phenomenon that is well known to social scientists; people zealously resist challenges to their most strongly held beliefs. Moreover, studies show that threatening those beliefs has the effect of people clinging to those beliefs even more fervently; the so-called “worldview backfire effect.” Thus, the stronger your evidence-based arguments against the vaccine-autism link might be, the stronger your disputants might cling to their anti-vaxxer position. The reason is the same as that which makes religious and political zealots immovable. See Aside 3.]

[Aside 3: Moses Maimonides (1138-1204), who many consider to be the greatest Jewish philosopher, confronted dogmatists in the 12th century, when writing his Guide to the Perplexed; his attempt to reconcile the Old Testament bible with what he considered to be the irrefutable scientific worldview put forth by Aristotle and other eminent Greek philosophers. In brief, Maimonides argued that the bible should not be taken literally but, instead, should be read metaphorically. Then, it could be entirely consistent with the truths arrived at through science and reason. Yet, Maimonides realized that most people did read the bible literally, and that to challenge their traditional point of view would be equivalent to challenging their faith itself. Thus, he realized that his arguments would be listened to by only a small group of the most open-minded readers.]

University of Sussex social anthropologist, Melissa Leach, suggests that scientists need to be more empathetic to the personal and cultural beliefs that cause people to reject scientific evidence (3). To that point, scientists need to listen to and understand the reasons why denialists seek alternatives to science, before they might be heard in turn. And scientists must be careful not to imply to science deniers that they are ignorant or irrational (see Aside 4). “Dismissing public and political concerns about health interventions as unscientific, irrational or misled fails to do justice to the different perspectives in play… It is why we see backlashes to even the best-intentioned initiatives (3).” In addition, scientists should not fall into the trap of advocating for an abstract principle. If you are perceived as an advocate for a policy, you may lose trust as an unbiased knowledge broker. So, stick to the evidence. Patiently and clearly connect the dots.

[Aside 4: It may surprise some that science denialists do not sort cleanly along income or education demographics. For instance, the movement to forgo vaccinations has become popular in some more liberal and affluent communities; the organic grocery demographic. Also, consider the example of conservative columnist George Will; an obviously well-educated and sophisticated individual, who nonetheless steadfastly maintains that since climate change happened naturally in the past, we cannot know that human-caused carbon pollution will cause harmful climate changes in the future. Others have noted that Will’s logic is equivalent to saying that since nonsmokers died of lung cancer in the past, we cannot know that cigarette smoking is a cause of lung cancer now. George Will also is not moved by the fact that there is a consensus among climate scientists—based on the accumulation of massive evidence—that human-caused carbon emissions are changing the climate. Climate scientists are now as certain of that conclusion as biomedical scientists are that cigarette smoking causes lung cancer.]

Better communication with science denialists is not easy for reasons noted above. Moreover, many science denialists have learned to rebut the consensus view by cherry-picking “scientific” evidence that might cast doubt on the consensus view; irrespective of whether their selected evidence came from poorly conducted experiments. Moreover, denialists may throw their “alternative facts” at you so fast that, in refuting them, you exhaust your energy and patience well before you get to make your own argument (see Aside 5). And there still will be vociferous politicians, who will continue to misrepresent and ignore science, to advance their own agendas.

[Aside 5: To that point, in 2013 Italian programmer Alberto Brandolini put forward Brandolini’s law (also known as the “Bullshit Asymmetry Principle”). It states: “The amount of energy needed to refute bullshit is an order of magnitude bigger than to produce it.”]

In early February 2017, scientists across the United States began to plan a March for Science, to take place in Washington on April 22; Earth Day. Are organized marches an effective way to promote a pro-science agenda? Some scientists say that the march might be counterproductive. For instance, Geologist Robert Young, of Western Carolina University, argued that the march “could deepen the divide between conservatives and liberals, reinforce the idea that scientists are a political interest group…There’s a section of the American electorate—whether we like to acknowledge it or not—that has become skeptical of science. . . I don’t think that scientists standing in Washington, giving speeches and holding signs, is going to convince those people that they need to pay attention to our concerns… Somehow, as a community, those of us who care about science need to find a way to communicate with those folks…It has to be direct communication or ways that we have not imagined yet (4).”

Young’s remarks provoked a notable backlash on Twitter, with most scientists coming out in favor of the march. Also, consider the outcome of a 2012 march in Ottawa, by Canadian scientists opposed to the anti-science policies of Canada’s conservative Harper government (Aside 5). The Canadian march did not diminish the credibility of the participants, nor did it lead to polarization of the public. Instead, by bringing the Harper government’s anti-science policies to the public’s attention, the march may have helped to elect the more pro-science government of Justin Trudeau in 2015. So, one might hope that an American march might have a positive effect here, even if only to stem the tide of misinformation being fed to the American public.

[Aside 5: Canadian scientists protested the Harper government’s restrictions against free communication between scientists and the media; particularly communications that opposed the government’s pro-industry environmental policies. Scientists who did not comply with the Harper government’s restrictions might have their research programs terminated. In the U.S., in December 2016, then President-elect Trump asked the Department of Energy for the names of career employees and contractors who attended U.N. climate talks over the past five years. He also requested emails of those meetings. The DOE responded with a statement saying that Trump’s request had “unsettled” many in its workforce, that the DOE would “be forthcoming with all [publicly] available information,” but that it would withhold “any individual names.”]

There is no middle ground between objective science and unsubstantiated “alternative facts.” As stated most eloquently by Wendy Palenfeb: “Evidence and objective reality are the foundation of successful policy and governance. Openness is as vital to science as it is to democracy. We cannot allow hard-won knowledge to be ignored or distorted (5).”

References

  1. President Trump’s Advocacy of the Debunked Link Between Vaccines and Autism, Posted on the blog January 26, 2017.
  2. Charles Sykesfeb, Why Nobody Cares the President Is Lying, NY Times, February 4, 2017.
  3. Melissa Leach, Accommodating dissent, Nature 450, p283, 22 November 2007, doi:10.1038/450483a.
  4. Diana Kwon, Will a March Help Science?, The Scientist, February 2, 2017.
  5. Wendy Palenfeb, When Canadian Scientists Were Muzzled by Their Government, NY Times, February 14, 2017.]

Addends

  1. Prompted by President Trump’s comments asserting a link between vaccines and autism, on February 7, 2017, more than 350 medical and professional organizations sent the President a letter stating that vaccines are a safe and most effective means for protecting the health of children and adults and saving lives. The text of the letter, and its signatories, can be accessed from: The week in science: 10–16 February 2017. Nature 542, (16 February 2017) doi:10.1038/542276a.
  2. The following is from an April 11, 2017 editorial in Nature, Nature supports the March for Science: “Finally, to the critics, yes it is true that the march blurs the lines between science and politics. But that line is already much fuzzier than some try to argue. It is possible to care about science and scientific thinking while ignoring the political context in which it operates. But it is difficult to do that and demand change at the same time.”