Luc Montagnier, the French virologist who won the Nobel Prize in medicine and Physiology in 2008 and was known for his discovery of the human immunodeficiency virus, died on February 8 at the age of 89.

Montagnier is a man of great achievement and great controversy. Although he made remarkable achievements in medical science, he was criticized for his unscientific opinions in his later years, including artificial theories on the origin of COVID-19.

In 2009, he claimed that DNA emits electromagnetic radiation, and that some bacterial DNA continues to emit signals long after an infection has been cleared; In 2012, he spoke at a conference on autism in Paris, France, claiming that long-term use of antibiotics can successfully treat autism; Last May, he claimed in a video that the vaccine programme was “unacceptably wrong” because it could cause the virus to mutate; In January, he criticized U.S. President Joe Biden’s vaccine policy in a Wall Street Journal op-ed with a Yale Law professor.

An accomplished scientist can be “incredibly” amateurish, confusing and incomprehensible. Fortunately, just because he is an “authority” does not automatically give his nonsense the property of truth.

Today, we send Off Montagnier in the 2020 edition, hoping to remember and thank him for his remarkable discoveries. Hopefully, one day AIDS, a hitherto intractable epidemic, will come to an end.

Article | Wang Chengzhi li Runhong
Director | Di Li Hui Tang Peilan

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Figure 1 2008 Nobel Prize in physiology or medicine for HIV and the discoverer of the HPV virus | photo source: nobelprize.org

On June 5, 1981, the Centers for Disease Control and Prevention (CDC) reported that five los Angeles-area gay men had developed pneumocystis carinii pneumonia.

The disease is very rare and usually occurs only in people with extremely low immunity. Soon, the CDC found more patients with similar symptoms, including Kaposi’s sarcoma and giant cell infections.

At the time it was called “gay man’s disease”. But it soon became clear that all groups of people caught the disease, mostly through sex or blood transfusions. As a kind of infectious disease, it is urgent to find its pathogenic factor to control the disease.

In the race to discover the cause of the disease, Luc Montagnier, a Frenchman, found and proved that HIV was the killer. His discovery, along with a dispute between him and another American scientist, has led to high-level government involvement and even court battles in France and the United States.

01

Montagnier, the refugee

Montagnier was born in 1932 in Chabris, a town in central France known for its goat cheese and white asparagus, among other produce. As an only child, Montagnier was naturally doted on by his parents and could have had a carefree childhood. But life’s accidents and the currents of The Times came one after another to disrupt Montagnier’s peaceful life.

At the age of five, Montagnier was hit by a speeding car on a main road. Montagnier spent two days in a coma before being revived. Although he escaped death, the accident left him badly injured and scarred.

Figure 2 5 montagnier is | photo source: Nobel Prize’s official website
Two years later, in 1939, seven-year-old Montagnier was picking grapes with his family in his uncle’s vineyard when France declared war on Germany. The next year, with the full invasion of Germany, the Montagnier family fled as refugees. Since then they have not enough to eat and often suffer from hunger. During the four years of the war, Montagnier, who was still in his teenage years, did not gain any weight.

To add insult to injury, Montagnier’s grandfather was diagnosed with colon cancer and died after enduring great pain. Seeing how his grandfather suffered inspired Montagnier to study medicine and cancer.

In high school, Montagnier kept his grades at the top, and his interest in science grew. He set up a simple chemistry lab in the cellar of his home and made his own hydrogen and scented aldehydes, even starting to make his own nitro compounds. At the same time, he also read a lot of books on physics, especially in frontier fields such as atomic physics.

After attending university, he decided to pursue medical research because of his strong interest in it. But there was no medical profession in his town at that time, so he went to the hospital in the morning and attended botany, zoology, and geology at school in the afternoon.

Fortunately, Professor Pierre Gavaudan, who teaches Montagnier’s botany, is not constrained by textbooks. His lectures on the latest advances in biology, including the DNA double helix, protein synthesis using ribosomes in vitro systems, and the structure of viruses, opened new doors for Montagnier.

Montagnier received two gifts from his father: a time-lapse camera and a microscope. He used them to study how algae cells adjust chloroplast arrangement in response to light intensity. Using different colour filters, time-lapse photography and microscopy, he showed that instead of using chlorophyll to absorb red light, the algae cells indirectly regulate the direction of the chloroplasts by using yellow pigments to absorb blue light. Montagnier, who was only 21, completed his Faculty of Sciences of Poitiers defense with this result.

02

First foray into the world of viruses

At 23, Montagnier became a research assistant at the Sorbonne Universite in Paris, where he learned cell culture techniques. In 1957, Heinz Fraenkel-Conrat and other scientists discovered that tobacco Mosaic virus uses RNA as genetic material and can infect plants through RNA. Montagnier was impressed and decided to study viruses through the then nascent field of molecular biology.

He then went to Kingsley Sanders’ laboratory in Castleton, England, to study the foot-and-mouth disease virus, where he showed that RNA could be copied by base pairing just like DNA.

Soon after, Michael Stocker, a renowned biologist, set up a new virus institute in Glasgow. Montagnier also came to study polyomavirus, a virus that causes tumors. Montagnier and his collaborators showed that the virus’s DNA itself has the ability to cause tumors. While this may not seem unusual to modern eyes, at the time it broke the conventional wisdom that all life depended on protein.

After his research studies in the UK, Montagnier returned to France to continue his research on oncogenic viruses at the Curie Institute. At the time, one of the great mysteries of virology was waiting to be solved: How did RNA viruses replicate? Using Rous sarcoma virus (RSV) as a model, Montagnier tried to identify the double-stranded RNA produced by viruses. He did find it in cells infected with the Rous sarcoma virus (RSV), but the sequence was not of viral origin. He also found double-stranded RNA in cells free of viral infection, proving that this double-stranded RNA was not related to the virus. Montagnier later recalled that the discovery was actually a partial approximation of the small interfering Rnas that were later discovered.

In 1970, across the ocean in the United States, two laboratories (Howard Temin and David Baltimore) independently discovered reverse transcriptase in RSV. This enzyme can use viral RNA as a template to make the corresponding DNA. This not only explained the replication of RNA viruses, but also broke the central dogma of biology that RNA can be made from DNA templates, and led to the realization that transcription can also be done in reverse.

Based on this, Montagnier discovered that the DNA retroscribed by the virus can be integrated into the genomic DNA of the host cell. Since then, retroviruses have become the focus of Montagnier’s lifelong scientific research.

In 1972, Jacques Monod, director of the Pasteur Institute in France, approached Montagnier to establish a center for virus research at the Institute. At the time, a heavily funded virus cancer program in the United States had made tumor-associated viruses a hot topic, and Montagnier had created the Viral Oncology Unit at the Pasteur Institute.

In 1973, J. C. Chermann and his collaborator Franoise Barre-Sinoussi joined Montagnier’s research center. Barre-sinoussi is experienced in detecting reverse transcriptase activity. Montagnier persuaded them to work together on retroviruses that cause cancer in humans. In 1977, they began testing blood and tissue collected from patients in Paris hospitals to see if they could detect retroviruses.

At this point, montagnier’s experiments were greatly aided by research in two other LABS.

Because virus replication in the body is blocked by interferon, Montani speculates that blocking interferon would make it easier to detect the virus. Just then Ion Gresser’s laboratory in Villejuif, France, produced an antiserum that neutralised interferon. In tests, Montagnier and Barre-Sinoussi et al. found that treating cells with this interferon antiserum increased the expression of endogenous retroviruses by a factor of 50. This greatly increased the sensitivity of their experiments.

At the same time, The Laboratory of Robert Gallo, an American virologist, invented T cell culture technology. By adding a growth factor (TCGF, then called interleukin-2) to the culture, human T cells that would otherwise be difficult to grow in vitro could be expanded.

With the help of these two approaches, Montagnier et al. began studying retroviruses in T cells taken from breast cancer patients.

However, just as Montagnier’s lab was preparing various techniques for studying retroviruses, a new retrovirus, HIV, began to spread among humans, with serious consequences.

03

Between the square inch, the sky is very different

After June 1981, the United States Centers for Disease Control and Prevention reported increasing cases of AIDS. What is the cause of this terrible disease? This became an urgent problem at the time.

Gallo speculates that the new disease may be caused by a retrovirus. Gallo, a pioneer in retrovirus research, has good reason to suspect so.

His lab not only invented T-cell culture, but also isolated the first human retrovirus, human T-cell leukemia virus-1 (HTLV-1). The symptoms of htLV-1 infection in animal models are similar to those of the new disease, and htLV-1 can be transmitted through blood transfusion, sex, or mother-to-child transmission. These clues led Gallo to believe that the new disease was caused by a retrovirus similar to HTLV-1.

In 1982, Gallo’s lab detected DNA sequences similar to HTLV-1 in blood samples from patients with the new disease. In early 1983, they found HTLV-associated DNA in two of 33 samples. It turned out that this was because both patients were infected with HTLV at the same time, but the experiments misled Gallo into thinking that the pathogen of the new disease was a new member of the HTLV family.

As it turned out, Gallo was right that the new virus was a retrovirus, but not a new member of the HTLV family.

Gallo’s conjecture spread through the field, with some clinicians and immunologists. Montagnier learned of these suspicions, too, and in 1982, he teamed up with his student Franoise Brun-Vezinet to study new pathogens.

On 3 January 1983 brun-Vezinet obtained a lymph node sample from a patient numbered BRU. Montagnier himself grinded the sample and grew the T cells in it. Fifteen days later, Franoise Sinoussi detected reverse transcriptional activity in the supernatant of the culture medium, confirming that the new pathogen was indeed a retrovirus. They called the strain lymphadenopathy associated virus (LAV).

At the time, THE HTLV-1 and HTLV-2 discovered in Gallo’s lab were the only known human retroviruses. Using Gallo’s HTLV antibody, Montagnier’s lab tested their samples and found that the HTLV antibody did not bind to their newly discovered virus. This suggests that the new virus is not HTLV, as Gallo had speculated.

Luckily for Montagnier, the first sample led him in the right direction. Later, he recalled, they received a sample of a patient named MOI infected with both the new virus and HTLV. If this is the first sample they test, they are likely to be as misguided as Gallo.

Meanwhile, Montagnier’s collaborator, electron microscopist Charles Dauguet, also found that the new virus looked different from HTLV under electron microscopy. Virologist Edwald Edlinger suggested that Montagnier compare the new virus with lentiviruses found in animals. Using electron microscopy, they found that the new virus and lentiviruses looked almost identical.

Figure 4 HIV | photo source: pixabay.com

Montagnier and Gallo shared their findings, and Gallo suggested a joint publication. In May 1983, Montagnier and Gallo simultaneously published their own isolates of the new virus. However, Gallo did not know at this time that the virus he had isolated was actually a mixture of the new virus and HTLV, and that Montagnier’s LAV was a pure strain. This is probably the main reason gallo was later unable to share the Prize with Montagnier.

Between late 1983 and early 1984, Gallo lab technician Betsy Reed-Connole and collaborator Mika Popovic isolated several new strains (RF, IIIB, and MN). During this time, they published a series of papers detailing their isolation methods, in vitro amplification methods, and evidence that the virus was the cause of a new disease. These strains also became the standard for subsequent blood tests.

At this point, Montagnier’s experiment ran into problems. The virus (LAV) they isolated from BRU patients could not amplify in T cells. Gallo visited Montagnier’s lab in Paris and brought him samples of the IIIB strain, which can proliferate continuously in T cells. Gallo wanted Montagnier to compare the IIIB and LAV strains, and they agreed to hold a joint press conference if the virus they isolated was the same.

04

The controversy has rattled the French and American top brass

Somehow, however, the news leaked out. A reporter contacted Margaret Heckler, then the secretary of health and Human Services, prompting her to urgently recall Gallo and hold an early unilateral press conference in the United States.

In May 1985, the U.S. Patent and Trademark Office granted gallo’s research institute a patent for a blood test for the new virus. In fact, France had already filed an almost identical patent application before Gallo.

Between 1985 and 1986, French representatives launched four lawsuits and two years of wrangling between the two governments. Then, in 1987, President Ronald Reagan and French Prime Minister Francois Mitterrand signed a settlement that offered shared benefits. The deal, however, gave the United States more rights until it was renegotiated in 1994 and the two sides agreed on a 50-50 split.

The hastily called press conference in the United States not only caused a political storm, but also generated a lot of debate in the scientific community. Coincidences since then have intensified the debate.

Montagnier isolated a proliferative strain called LAI after LAV. Possibly because this strain of virus is highly proliferative, it contaminated montagnier’s previous LAV samples. Montagnier had sent this sample to Gallo, which led to the contamination of gallo’s lab strain. Gallo later found that IIIB isolated in his laboratory was also contaminated with LAI, and the contaminated strain spread to many laboratories. Although the contamination between these strains was gradually recognized in the 1990s, it did not affect scientists’ understanding of their cause of immunodeficiency disease in humans.

Figure 5 December 10, 2008 montagnier won the Nobel Prize in the Stockholm concert hall | photo source: nobelprize.org

After academic discussion, the new virus was finally named as human immunodeficiency virus (HIV), and the diseases it causes are collectively known as immune deficiency syndrome (AIDS), so HIV is also referred to as HIV.

Despite many twists and turns, Montagnier’s contribution to the first isolation of HIV has been recognized by the academic community. Gallo’s commentary, published in Science in 2002, also explicitly acknowledged that Montagnier’s team was indisputably the first to isolate true HIV, since his team had failed to exclude HTLV from the strain published in 1983.

In 2008, Montagny and his collaborator Franoise Barre-Sinoussi shared the Nobel Prize in Physiology or Medicine for their discovery of HIV. He later became one of the co-founders of the World Foundation for AIDS Research, contributing greatly to the understanding of HIV and AIDS.