The lab’s research into human retroviral pathogenesis was going equally well. Judy was working on successful projects as a postdoctoral fellow, such as developing the first molecular clone of HTLV-1, which was useful in determining the gene expression required for malignant T-cell growth, and the role of the naturally occurring defective (incomplete) viruses during viral infection. As a technician, her projects included the role of methylation (which alters gene expression) in retroviral life cycles; anti-viral mechanisms of type 1 interferon; and other cytokines. Judy and Dave Derse gave presentations at the 1992 Cold Spring Harbor RNA Tumor Virus meeting on the characterization of these defective viruses.
Scientific progress seemed to be running smoothly, but in the late 1990s, a turn for the worse was brewing. Derse was becoming irritated that Judy was wearing two hats. Remember all of her salary was being paid by my lab budget and she still had to conduct scientific studies outside of her postdoctoral studies to receive her salary. Thus, she was not able to be fully dedicated to his instructions. But he was able to recruit two other fellows who were. I always felt that Derse did not want to share the HTLV-1 podium with me and that was the root of the problem.
Whatever the reasons, Derse made enough loud and unfair complaints about being in my lab that he received permission to find a new lab. While these events were occurring, Derse had submitted a paper on defective viruses with Judy as senior author. After Derse left my lab, he took Judy’s name off the paper, a thoroughly unethical act. In science, credit and publication are the coins of the realm, and nobody needs it more than a junior researcher.
I was completely disgusted by this theft of Judy’s work by Derse.
***
I’d published many papers concerning the regulation of blood cell development, concluding with Steve Bartelmez’s paper on regulation of stem cells with TGF-beta.6 The use of the anti-TGF-beta antibody was a crucial reagent in the success of any investigation.
I got a call from a former colleague, now working at Biogen, who told me that Biogen had bought the antibody, and I was now forbidden to use it in my research. I then produced a signed material transfer agreement which allowed me to use the antibody. I told him further that the antibody producing cells were available from American Type Culture Collection, for all investigators to use.
The cells were removed from the American Type Culture Collection within forty-eight hours after my phone call. I was then informed I would get no more TGF-beta reagents, particularly the humanized antibodies they were making.
So, at a key point, after almost a decade of studies, we needed to find a new technology, particularly to translate our findings from mouse studies to humans. Bartelmez began a wonderful collaboration with Pat Iversen, who was pioneering a new technology using phosporodiamidate mortpholino oligomer (PMOs). PMOs are safe, synthetic antisense molecules which bind to RNA molecules in a sequence-specific manner to prevent a given protein from being made. He made several PMOs against TGF-beta that were very efficient.
Pat Iverson, using PMO antisense molecules as antivirals, has shown that PMOs were very effective against the lethal Ebola virus and SARS-1 coronavirus, as well as several other viruses.
Why nobody was listening and didn’t try it against COVID-19 is beyond me and a clear example of the myopic nature of science.
The success of the Derse revolt emboldened other members of the lab to voice that they wanted to leave the lab. The report from the 1998 site visit contained stunning requests to leave.
It was stunning because none of these people ever told me they were unhappy. The request from a female investigator was a shock because I had spent lots of time giving her the ability to get tenure, when the site visit itself had recommended eliminating her position. Ironically, the only investigators who did not complain were the ones who had performed excellently in the site visit. To this day, I do not know what I did so wrong to warrant the horrible reviews from those I’d attempted to help.
Furthermore, three other scientists moved on from the lab to have very good scientific careers.
Well, at least hard-working clever Judy didn’t leave the lab, at least not voluntarily. She was forced out by several people, the most vociferous of which were the Derse lab members who stole her work.
Judy published a seminal paper that HIV infection upregulated DNA methyltransferase, which caused de novo methylation of the gamma interferon gene, which downregulates the production of gamma interferon.7 This mechanism is one way the virus evades T-cell immunity. Furthermore, she showed that infection with an HIV virus which could not integrate, and therefore could not be infectious, was still able to stimulate de novo methylation, suggesting that a virus can cause pathogenic changes without productive infection.8
In January 1999, Judy was made head of the Lab of Antiviral Drugs Mechanisms. I felt it was done because it was thought that separating us would make us both weaker. I believe the NCI hierarchy fully expected and wanted her to fail. They also believed it would make my lab weaker and less creative.
One of Judy’s hires at the Lab of Antiviral Drug Mechanisms was from Shanghai and did not want to take orders from a woman. Judy would write up the experiments she needed to be done, and then I’d give them to the doctor, who would do the work.
Needing to find a replacement for Judy, I hired an experienced retrovirologist who was working in Baltimore. Gallo was coming to work at her university, and she wanted to get out of there as quickly as possible.
With the cloning of the chemokine receptors, it became obvious that chemokine receptors were a cofactor for HIV cell entry. Peptide T specifically inhibited viral strains which used CCR5 receptors for entry into the macrophage, microglia, and CD4 T-cells, but had no effect on lab-adapted strains using CXCR4. This was vindication we were right.
But as usual, nobody was listening. We hoped there would be other applications for this reagent.
***
Being absolutely sure that Peptide T would be a useful reagent, I wrote a grant between Candace Pert, Michael Ruff, and their company to study how to make Peptide T a commercially viable product.
With reviews straight out of 1986, my request to study Peptide T was denied.
In 2001, Judy permanently left Frederick, the NCI, and the Lab of Antiviral Drug Mechanisms. She accepted a scientific research position with EpiGenX Pharmaceuticals in California. This move allowed her to pursue her three loves; methylation, viruses, and David Nolde, whom she married on October 7, 2000. Her instincts were right about methylation.
It is my feeling that epigenetic therapy to eliminate the pre-metastatic niche before tumor cells can use these niches to metastasize to other body parts will be the next major breakthrough in cancer treatment. One of the mechanisms involved is fascinating to us. The epigenetic therapy induced endogenous retroviruses expression which stimulated the production of interferon-alpha in the host, which is part of the immune response to eliminate the pre metastatic niches. That is to prevent the cancer from spreading.
For her farewell party, several of her close girlfriends and me (the designated driver) celebrated at Tauraso’s restaurant (our usual spot for lab Christmas parties). Nick Tauraso, a former virologist turned sommelier/restaurateur, paired excellent wines with food. I drove all the women home, with Judy being the last stop.
As I was walking to my car, I heard a loud crash, which made me turn around. Judy had fallen over the railing and landed on top of the Japanese maple, recently planted by her brother-in-law. I rescued her and made sure she got to her apartment without further incident.
All of her belongings had already been shipped to California, so that next day she flew out wearing the same pair of jeans she’d worn the previous night, which now had a large rip in them. We never told Judy’s brother-in-law how his Japanese maple had been destroyed.
A result of my laboratory’s next site visit in 2002 was the recommendation that my laboratory be closed. Maybe it was inevitable. Three of the four independent investigators had de
manded to leave the lab and the fourth had already left the lab.
It was easy for the site inspectors to claim the lab was in decline. But from my perspective it was a different story, and one all too common in government. Having two hands tied behind my back, I have never felt I was given a fair chance to succeed. It was a bitter pill to swallow, considering the years I’d dedicated to government service.
CHAPTER NINE
Millennium Changes
I worry that, especially as the millennium edges
nearer, pseudoscience and superstition will seem year by
year, more tempting. The siren song of unreason more
sonorous and attractive.
—Carl Sagan1
Two decades into the millennium, several studies have demonstrated that when compared to other developed countries, more Americans prefer dictatorship over democracy.
There could be many reasons for this, but one is the complete failure of the public health apparatus, the FDA, the CDC, the Environmental Protection Agency (EPA), NIAID, NIH, and the HHS, who are charged with making the regulatory decisions to protect our public health. Their mixed messages in any crisis, the many decisions that enrich their friends and regulators, and the utter contempt with which they treat the public lead to a crisis of confidence. Most citizens, including those who research these issues deeply, do not know who to believe.
As I reached another crossroads with my laboratory torn asunder, could I continue and rebuild my lab again? With all the corruption, I knew that no matter what, I must live my life within the bounds of my value system. My obligation was to continue to simply focus on the science as I have always done.
Luckily, as long as I had a budget, I could work on projects without having to justify the funding, at least until the next four-year program review. Elsewhere, as a Nobel laureate wrote on the 2014 death of Fred Sanger, a two-time Nobel Prize winner:
A Fred Sanger would not survive in today’s world of science. With continuous reporting and appraisals, some committee would note he published little of note between insulin in 1952 and his first paper on RNA sequencing in 1967, with another long gap until DNA sequencing in 1977. He would be labeled as unproductive, and his modest personal support would be denied. We no longer have a culture that allows individuals to embark on long term and what would be considered risky projects.2
Most of the scientists who perform experiments agree with me that this change in culture was led by the rise in dominance of the bureaucrats/physician-bureaucrats who are themselves fundamentally unproductive, and that this is a serious long-term threat to science. However, the correct use of science can still be a “candle in the dark.”
The exploration into uncovering the causes which are sure to be multi-factorial in the initiation and progression of chronic diseases has always intrigued me. I would continue to investigate the role of the innate immune system in the pathogenesis of retroviruses, particularly the cells which regulate type I interferon, the body’s natural antiviral, and the use of transient inhibition of TGF-beta in stem cell therapies in chronic diseases.
The concept that macrophages could support productive, restricted, and latent expression from Judy’s thesis persuaded us that we needed to examine the innate immune system to fully understand pathogenesis and develop therapeutic strategies. The innate immune system (consists of rapid physical, chemical, and cellular nonspecific defenses against pathogens) is the first line of defense against invading pathogens.
In the innate immune system, sensors like pattern recognition receptors (PRRs) detect specific viral components such as viral RNA, DNA, or intermediate products which stimulate production of interferon alpha/beta and activate other immune cells. Recent studies show interferon lambda directly stimulates restriction factors that inhibit HIV replication and also upregulates intracellular expression of type I interferon. It also shows that interferon has both extracellular and intracellular antiviral mechanisms which differ. To use interferon alpha therapeutically, one must control the potential detrimental effects.
Never having been comfortable with the fact that cell-free HTLV-1 virions are poorly infectious in vitro, I asked my new staff scientist to examine infectivity in innate immune cells. She showed HTLV-1 could efficiently infect myeloid and plasmacytoid dendritic cells (DC) and even more importantly, could rapidly and efficiently transfer infectious virus to autologous primary T-cells which became infected.3
We also found that both types of dendritic cells could act as a true viral reservoir. The first cells to be infected in vivo.4
These studies suggested to us that impairment of the dendritic cells function after HTLV-1 infection plays a role in pathogenesis.5 Emphasis was placed upon HTLV-1 induced alteration of type I interferon shown by several investigators. It was our hypothesis that the development of acute T-cell leukemia (ATL) is a struggle within the body between HTLV-1 and type I interferon production. The HTLV-1 p30 protein is required for infectivity in vivo and for dendritic cells in vitro, but not for T-cells.
The p30 protein was shown to dampen interferon responses in monocytes and dendric cells, representing an early step for HTLV-1 infection. Interferon type I and AZT treatment of acute T-cell leukemia can result in complete remission.6 Again, this suggests that exogenous type I interferon, in contrast to endogenous interferon type I, can act therapeutically in HTLV-1 infection.
What about other viruses?
Kaposi’s sarcoma associated herpes virus, in which B lymphocytes are the major infected reservoir in living organisms, cannot be infected by the virus in lab culture. We demonstrated that the dendric cells mediate infection of primary B-cells.7 We point out in the interferon chapter in Part Two that, for some unknown reason, Fauci, despite praising interferon use in AIDS patients and NIAID running a successful trial in 1989,8 failed to recommend interferon and AZT trials when AZT was the only approved drug and obviously was working alone.
Why is Fauci again ignoring using interferon for COVID-19, where an imbalanced host response, defined by low levels of type I and type III interferons (antiviral defenses), next to elevated chemokines and IL-6 (inflammatory marker) are driving the viral inflammatory response? It is clear as several papers have shown that there are benefits to exogenous interferon treatment using either interferon alpha or lambda.9
The second project involved a collaborative effort between Steve Bartelmez, Pat Iverson, and me, involving the ex vivo inhibition of endogenous TGF-beta in purified hematology stem cells (HSC). The astonishing results permitted complete functional transplant with as few as sixty HSCs, instead of a thousand cells, which could rescue mice from lethal irradiation.10
Both the anti-TGF-beta antibody and PMO antisense TGF-beta treated HSCs produced accelerated engraftment, resulting in high levels of donor cells, and a durable graft. Also, never seen before, the early donor cells were neutrophils, which protect a transplant against infection. This was truly miraculous and paradigm-changing.
After twenty years, we can still not get it published in a good journal because the reviewers don’t believe it. It’s not religion. It’s science. It was important enough that I wanted to compromise with the reviewers to get it published, but Steve never wanted to do this. Without telling me, Steve submitted it to a journal (Stem Cell Research and Therapeutics), which few could ever read because it is not available online.11 If a scholar’s obligation is to share knowledge, in this instance we have failed dismally. It is published, but nobody is reading it.
To continue these cell therapy studies, we needed to show that this therapy would work on human cells. Steve wisely chose diabetic retinopathy, a common cause of vision loss and blindness. Healthy human HSCs can go to the retina and help repair vascular leaks, but HSCs of diabetics cannot do this vascular repair. We demonstrated that human specific PMO antisense TGF-beta can reverse the dysfunction of diabetic HSC so that they are now able to repair the endothelium in the retina.12
Since the only clinical therapy is anti-VEGF, which fails 50 p
ercent of the time, I believe this additional therapeutic approach should be an attractive approach for those patients who need a therapy. I have a great deal of frustration that none of these observations have led to clinical trials despite a decade of effort.
I believe ex vivo manipulation of stem cells have a bright future, if the funds are provided for the appropriate investigations.
***
Many of my collaborators and friends seemed to be passing out of my life, and I didn’t understand why.
One of the most lamentable losses was Dr. Kendall Smith. I always praised his world-class work on the biochemical mechanisms of IL-2, which he originally called “Frankie’s factor.” He often invited me to his lab and I’d stay in his home, having long conversations about science and enjoying his wife’s amazing French cooking.
One of the best AIDS meetings we ever attended was in Reims, France in April, 1995. It was the first AIDS meeting that acknowledged the importance of cytokines in HIV pathology and treatment. Kendall knew French cuisine, so Judy, Kendall, and I went to an expensive restaurant during the conference, at which the Prix Fix meal was an all-truffle menu. None of us knew that the menus left the last digit off the prices, and that the women’s menu had no pricing at all.
Knowing Judy was still a graduate student on a technician’s salary, Kendall and I tried to eat less expensive meals and give Judy the opportunity to try the truffle menu. Even with the truffle ice cream, the price appeared to be $150.
Judy gave Kendall her American Express card to pay for the bill and Kendall took care of everything since he spoke French fluently. When Kendall returned to the table, he seemed uncomfortable.
Ending Plague Page 13