Dr. Janet Nicholson

CHAMBERLAND: This is Dr. Mary Chamberland, and I'm here with Dr. Janet Nicholson at the Centers for Disease Control and Prevention [CDC] in Atlanta, Georgia. Today is Monday, August 14, 2017. I'm interviewing Dr. Nicholson as part of the oral history project, The Early Years of AIDS: CDC's Response to an Historic Epidemic. Jan, welcome to the project. Do I have your permission to interview you and to record this interview?

NICHOLSON: Yes.

CHAMBERLAND: Jan, your career at CDC spans some three decades. You came to CDC in 1979, so a year or so before the June 1981 publication of the first MMWR [Morbidity and Mortality Weekly Report] report on Pneumocystis carinii pneumonia among homosexual men. Your work in the laboratory of the late Dr. [John] Steven McDougal resulted in some of the early seminal discoveries and publications about the human immunodeficiency virus or HIV. You then went on to assume a 00:01:00series of senior leadership positions within CDC's Center for Infectious Diseases before retiring in 2013.

Before we explore your CDC work in more detail, let's talk a little bit about your background. Can you tell us where you grew up and about your early family life?

NICHOLSON: Sure. I grew up on a farm in Iowa, and I have five sisters. There were six girls, and my father was a farmer; to his chagrin, no boys. Then I went on to college about 20 miles away at Buena Vista College, it's now Buena Vista University in Storm Lake, Iowa. I got my bachelor's degree there in biology with a minor in math and general science, basically. I wanted to be a math teacher 00:02:00when I went to college, and when I first got into calculus one, two, and three and learned that I couldn't tutor others in that subject, I decided maybe math isn't for me. Biology was my other love, so I got a degree in biology. After I graduated, then I took a job in Omaha, Nebraska, at the Eppley Institute for Cancer Research, working for what we called a mad Hungarian who was doing research in basically hydrazine compounds and their effect on small laboratory animals. That was not a very happy job, because all we did was feed these animals compounds, chemicals and then monitor their health and do autopsies on them (or necropsies) on them. I got tired of that and tired of the cold in the 00:03:00Midwest and went south to Houston, where I had an aunt and uncle who lived in Houston, and applied for jobs in the Houston area.

The first job I was offered was in Galveston, Texas. I worked in the lab there of [Dr.] Alan Goldstein, who was doing research in thymosin. Thymosin is a hormone made by the thymus gland, and for which he was well known. He brought a research crew from New York to Galveston. Then they assembled more workers, and I was one of those that they assembled. I worked there for about three years or so and decided I wanted to go to graduate school. I had a friend who worked at Morehouse College who invited me to Atlanta to look at graduate schools.

I came to Atlanta and went to school at Emory University in the program, at that time it was called Experimental Pathology. They take two students a year, and it was actually the pathology department that sponsored that particular program. I started with Dr. Mariano LaVia, who was working on Fc receptors on B cells. I stayed there for a while. I finished my course work there. I was working in the lab all this time, as well on some research projects. Then when he took a job in Charleston, South Carolina, at the Medical College of South Carolina, I elected not to go with him. He made arrangements for me to finish my research, because that was all that was left for me, at CDC, and that's how I got in the door at CDC.

CHAMBERLAND: So, that's quite a story coming from a farm in the Midwest and ending up in the big city of Atlanta. Do you know what got you interested in science and math?

NICHOLSON: I really enjoyed biology in high school and that just clicked with me. I enjoyed biology in college, and that really clicked with me as well. I enjoyed the studying, the organisms and the systems, and all those things that go into life basically.

CHAMBERLAND: And you liked the whole idea of being a laboratory researcher, doing bench laboratory work?

NICHOLSON: I like working with my hands. I think that was the big thing too. I 00:06:00like doing things with my hands, and that was really helpful. I'm also fairly detail-oriented, so that all fit together to work in a laboratory in great detail.

CHAMBERLAND: So, when your mentor moved from Emory to Charleston and arranged for you to work on finishing out your Ph.D. affiliated with CDC, did that end up changing the focus of your research for your Ph.D.?

NICHOLSON: Yes and no. My mentor was very interested in B cells and the receptors on B cells. When I came to CDC, T cells were the focus. In that way, yes it did, but I think I was in a much better place working in the area that I 00:07:00was working, because it appeared that the story that he was trying to spin on Fc receptors didn't really pan out. Whereas the utility of the work on the T cell receptors was a lot more viable.

CHAMBERLAND: So, maybe we should have you, because I know this is going to come up, be a constant theme in our conversation today, and it might be good to take a step back and have you explain very briefly what B and T cells are. They are types of white cells, types of lymphocytes?

NICHOLSON: Right. There are a number of different kinds of white cells in the blood. One of those kinds is lymphocytes. About a third of the white cells in the blood are lymphocytes. There are basically three kinds of white cells: there are T cells, B cells, and natural killer cells. The B cells are the ones that 00:08:00make antibodies, so when your body produces antibodies, those are the cells that make it. T cells, there are very many different kinds of cells, but they're generally known as helper cells. They help the B cells make antibody, and they help in other kinds of immune responses. The natural killer cells are a much smaller proportion of cells, and they do as described. They are natural killer cells, nothing special about the things that they kill. They just are there.

CHAMBERLAND: So, combined the T and B cells sort of team up, if you will. Their function is to allow us to fight off, be it infections or cancers or whatever.

NICHOLSON: Exactly.

CHAMBERLAND: This is the late '70s, and this field of immunology is really still 00:09:00very new, nascent. Is that correct?

NICHOLSON: That's very true. We knew about B cells; we could identify B cells. We couldn't identify T cells. It was the absence of the markers or the antigens you would find on a B cell that made it a T cell or a natural killer cell. We didn't know about at that time either, but it would make it a T cell. There were some markers on cells that they thought were associated with T cells, but it wasn't until probably the late '70s that it was identified that there were special ways that you could identify T cells.

CHAMBERLAND: So, this is interesting because this sort of effort to unravel the secrets of the immune system and the properties and functions of these various cells is just starting to happen before, actually the first cases of what would 00:10:00become known as AIDS were being recognized in the United States. So, then when you switched over to finish off your Ph.D. at CDC, CDC was working on T cells and who was your mentor at CDC?

NICHOLSON: It was Steve McDougal. Actually, he was working on immune responses in mice. The group that we were with that became the Immunology Division, for a short time anyway, was really interested in leukemias. So, the vast majority of the people in that group were studying leukemia. Steve was an arthritis guy--

CHAMBERLAND: Immunologist.

NICHOLSON: --an immunologist, and the basic research that he was doing was in mice, looking at T cell subsets in mice. On the human side, it wasn't very long 00:11:00before the mouse work that the T cell antigens were beginning to be developed and then reagents to those markers were beginning to be developed. It was like a perfect storm in a way in the field.

CHAMBERLAND: A good storm.

NICHOLSON: A good storm in a good way. The technology was becoming available, the tools to use for that technology were becoming available, and the way of looking at things was all coming together about the same time in the late '70s, very early '80s.

CHAMBERLAND: After you tied up your Ph.D. thesis, did you want to stay on at CDC? Could that be arranged?

NICHOLSON: Of course. I spent about a year and a half on my thesis for my Ph.D., 00:12:00and it was looking at a mouse model. Then everybody wants to come to CDC, so I was actually offered a post doc [doctoral] position to do similar sorts of work in the lab with Steve McDougal. I engaged in that for about a year, but it was quite interesting. I told you 1980 was a particularly--

CHAMBERLAND: That was a big year for you.

NICHOLSON: It was a big year for me. I was pregnant with my first child, I was finishing up my Ph.D., and I was offered a job at CDC. This was all in September and October of 1980. It was interesting, because the way the federal government works, there's end-of-year funding. What they do at the end of the year is get 00:13:00everybody's wish list for things to fund, because if there was extra money, they would fund some things. They often looked for big-ticket items, and one of the big-ticket items that somebody had put on the list was an FACS machine, which was a fluorescent-activated cell sorter, which was a huge piece of equipment. Very expensive piece of equipment. That arrived in September of 1980 as well. When I had agreed to stay on at CDC, was offered to stay on at CDC, I was offered to learn how to run that piece of equipment.

CHAMBERLAND: It was ordered without an operator.

NICHOLSON: That's right. I was trained as the operator and went away to learn how to do that.

CHAMBERLAND: You had to go away?

NICHOLSON: I went to Mountain View, California, for a week to learn how to 00:14:00operate that piece of equipment.

CHAMBERLAND: The manufacturer sponsored these training sessions?

NICHOLSON: Yes. Part of the purchase included the training.

CHAMBERLAND: You were the designated FACS [operator], or flow cytometer, is that another name?

NICHOLSON: Actually, at the time it was called a cell sorter.

CHAMBERLAND: Cell sorter.

NICHOLSON: --because it was a unique piece of equipment that would do cell sorting. The term "flow cytometry" came along a little bit later, after those machines. There were machines that were developed that didn't sort cells but had the same kind of way of looking at cells.

CHAMBERLAND: I think I'm going to have to ask you again to give us a little bit of a primer on the FACS machines or cell sorters. They're sorting cells. What does that mean? What are they sorting them into and how, in a simplified explanation, is that done, if there is a simplified?

NICHOLSON: It's not simple, it's not simple. This machine actually took up a whole room. We had to have a room modified for it. It had two 4-foot lasers on it. It had a computer unit that was the size of a refrigerator. The lasers were water-cooled, so we had to have cooling water that was pumped into that room. The bench on which this sat fit those two 4-foot lasers and the front end of it, if you will, that did the work. What would happen is that it would analyze individual cells. It would take the cells through a little tube that shot it into a nozzle with a very small tip on it, so that the stream that came out of 00:16:00that nozzle was smaller than a hair, the diameter of a hair. What it did was, it caused the cells to flow through that nozzle, single cells, one at a time, and that stream then was interrogated by one of the laser lights. The way the laser looked at the cells was what was collected on the computer. All cells have different sizes, they have different granularity, and those characteristics diffract light differently. What we did was, we added antibodies that were labeled with a fluorescent compound to the cells. Those cells that reacted with those labeled antibodies, when they passed in front of the laser light, were excited by the laser light and then emitted a color, which was picked up by the 00:17:00computer as well.

CHAMBERLAND: Because the antibody was attaching, if you will, if it found a matching antigen on the cell surface.

NICHOLSON: Yes. On the cell surface.

CHAMBERLAND: It sounds like a very complicated process, but it helped you unlock some information, vital information about white cells.

NICHOLSON: Right. The advantage of the cell sorter is, number one, you don't have to look through a microscope and hunt for cells. You don't have to determine whether they're a little bit bright or a lot bright or somewhat bright, because the flow cytometer, or the cell sorter as it was called at the time, would tell you how much fluorescence the cells had. For most things that you labeled, there were cells that were not labeled, there were cells that were labeled, and you could easily distinguish those using the computer on the cell sorter. Now, it was called a cell sorter at the time because not only did it 00:18:00analyze those cells, but if you told it you wanted to sort only the cells that had a label on it, it would send an electrical charge to only those cells. Then the stream would be put in front of two magnets, and those charged streams would then be diverted because--it's very complicated. But there was a way in which little droplets were made, because they were shaking that nozzle in which the cells were coming through. The cells came out of the nozzle in droplets, and the droplets were then-- The cells that were charged went in one direction, and the cells that were not charged went in another direction. The little droplets that had nothing in them went straight into the waste receptacle. That was a way you could physically separate individual cells. We didn't really do that; you didn't 00:19:00have to do that for the studies that we did, we simply analyzed the cells, but [the machine] had the capability to do that.

CHAMBERLAND: So T cells, cells in general, have on their cell surface various proteins and whatever. I know we'll talk a little bit more about this. You and your colleagues at CDC and elsewhere were really at the cutting edge of trying to identify what some of these structures were on top of the cells, and it was through the use of these antibodies. Now, this is probably the last technical thing I'm going to ask you to explain because, again, it's so foundational to the research that ensued about early studies on the HIV virus, but you had to make these antibodies?

NICHOLSON: No. The antibodies were finally commercially available in the early 00:20:00'80s, about 1979, 1980, 1981; there were companies that were making these antibodies.

CHAMBERLAND: And these are the so-called "monoclonal antibodies"?

NICHOLSON: These are monoclonal antibodies. You could get them either as straight monoclonal antibodies, or you could get them with a fluorescent label on them. Or you could do what they call an indirect stain, and you could take the unlabeled antibodies and then add a labeled anti-antibody that's fluorescent stained.

CHAMBERLAND: It sounds incredibly tedious. Did it take a long time to run your experiments? I mean, it just sounds like--

NICHOLSON: Not really. To simply label the cells didn't take long at all. Generally, that was a 30-minute incubation. Preparing the cells was another issue. At that time, the way lymphocytes were prepared was to put all the blood 00:21:00then, anticoagulated blood, over what they called a density gradient. That was a gel-like substance that you layered the blood onto; you put it into a centrifuge, so all the things that went to the bottom were heavier. In that case, it was the red blood cells, because they had hemoglobin in them and it made them heavier. The white blood cells sat at the top of this interface. You harvested the cells off the top of this gel-like substance and washed them to remove the things that you didn't want. Then we incubated them with the antibodies and then prepared them for the flow cytometer.

CHAMBERLAND: As a non-laboratorian, I find this fascinating, but I'm just also really taken aback by the complexity and, as you say, the level of detail. But 00:22:00it's very foundational to understand the studies that followed. Now at the time, CDC was one of the few places, relatively few places, that had one of these cell sorters?

NICHOLSON: There were probably about a dozen around the country at the time that we first got ours. They were mostly used for research. There were some that were used for cancer studies, but we were still learning more about the human immune system at that time.

CHAMBERLAND: They were apparently so relatively uncommon that I understand that when VIPs would come to CDC for tours of the facilities and laboratories, that they often were taken to the room where the cell sorter was. There's a very 00:23:00famous photo that appeared in the Atlanta Journal and Constitution of one of your more famous visitors, is that correct?

NICHOLSON: Yes. [President] Jimmy Carter came to visit us. This was after he was out of office; he was a fascinating guy. We gave him a tour of the cell sorter. You're right, a lot of higher-level people came to visit, including--I don't think this was in the Journal Constitution--- but it was [Ms.] Margaret Heckler, who was then the Secretary of Health and Human Services, came through. But, yes, we did a nice dog-and-pony show for a number of people.

CHAMBERLAND: All right, so now you've prepped us, you've armed us with this foundational background information. Let's get on to how you used it to uncover some of the secrets of this mysterious new disease. In June 1981 the MMWR comes out, this unusual opportunistic infection being diagnosed in homosexual men, 00:24:00initially in California and New York City. How did Steve McDougal's lab first get involved in investigating and doing studies about this new syndrome? It didn't have a name at the time. How did that happen?

NICHOLSON: That's a very good question. Because we were the Immunology Branch, specimens were sent to our branch to do some T and B cell work on them. At that time, that was in probably the summer of 1981. Between the time that we had received the cell sorter and that event, I had been working with another guest researcher that was doing work on infectious mononucleosis; we were looking at 00:25:00the subsets of T cells in mono. So, we had the technology available, we had the reagents available, and we were set up to do whatever came in the door related to HIV.

CHAMBERLAND: Oh, perfect, well timed.

NICHOLSON: The timing was great.

CHAMBERLAND: These specimens come in to CDC from these cases of this unknown disease, so you crank up the cell sorter and start running their blood cells through it. What do you observe? What are you finding?

NICHOLSON: We confirmed what the reports had reported, and that was that there were decreases in CD4 cells in these people.

CHAMBERLAND: These are the T helper cells?

NICHOLSON: These are the T helper cells, in these people with increases, concomitant increases, in CD8 cells--

CHAMBERLND: The killer cells--

NICHOLSON: --the killer cells, the cytotoxic suppressor cells.

CHAMBERLAND: This is bad, right? This is a bad thing.

NICHOLSON: This is a bad thing.

CHAMBERLAND: Because it affects your ability to fight off infections.

NICHOLSON: Right. Of course, then there was a whole lot more to know, because we were seeing people that were at the end stage of HIV infection. The HIV-positive AIDS patients, as we learned later, were those that we were testing. Of course, that spawned a lot of questions, certainly on the epidemiology side. This is where our big connection with epidemiologists came in, to learn more about the whole syndrome.

CHAMBERLAND: At the end stage, some of these individuals didn't have any T helper cells?

NICHOLSON: Very few, yes.

CHAMBERLAND: What was your reaction? What did people think? Had they come across anything like this previously?

NICHOLSON: No, this was something completely new. It was completely different, as far as I can remember, from anything else. With infectious mono, for example, there were increases in cells and not decreases in cells. That was-- it was very much different.

CHAMBERLAND: There's profound immunosuppression, if you will--

NICHOLSON: Exactly.

CHAMBERLAND: -- allowing these opportunistic infections to take hold, and this is what cases were succumbing to in those days, as you say, with end-stage AIDS. Before we talk a little bit more about some of the studies, I want to get a little bit of a fix on-- it sounds like a pretty small operation.

NICHOLSON: It was.

CHAMBERLAND: There's you and Steve McDougal. Are you working with anybody else at that time at CDC? Were there other immunologists or people? I'm just curious.

NICHOLSON: On the technical side, the technicians in the laboratory in the Immunology Branch or Division, as it became known for a short period of time anyway, all were very adept at doing the marker work, because they had done some of that for the leukemia studies that they had been in. [Dr. Thomas] Tom Spira and Steve McDougal and I, and to some extent [Dr. Steven] Steve Shore, a physician, were involved in the early HIV days. Steve Shore left not long after that, I believe, and went on to do infectious diseases in a hospital in Atlanta. So, it was a small group.

CHAMBERLAND: It really was a small contingent. I'm just trying to get a sense of what was the atmosphere like. You're getting these astounding results. Are 00:29:00people excited, dumbfounded? Are ideas popping out of your head about what you want to do next in terms of studies?

NICHOLSON: At the very beginning, there was such an onslaught of samples that came in that we were completely overwhelmed. Our lab actually expanded a good bit just to process all the specimens that came in. I told you the blood had to be put over a density gradient in order to separate the lymphocytes. The laboratory had great experience in freezing lymphocytes, and so a good portion of these samples was then frozen away for future studies. Then the flow cytometry was done on just a portion of that blood. It wasn't until we developed technologies or used technologies to remove the red cells without the density gradient that we were able to process much faster. It was several years of just 00:30:00an onslaught of specimen testing.

CHAMBERLAND: Is this because clinicians and researchers in other parts of the country are sending specimens to CDC, because you're just one of the few places that can do--.

NICHOLSON: We really didn't do it that way. The specimens that came to us were those that were through studies for the epidemiology side of the house.

CHAMBERLAND: Did that happen fairly quickly, that there was a joining up of the lab and the epi side of this early HIV research?

NICHOLSON: Yes. I remember that they started Task Force meetings, and a number of the studies were developed and described in the Task Force studies. We were invited-- we the immunology group-- were invited to come and participate in those biweekly meetings, I believe, to learn about what studies were coming and 00:31:00to report back on studies that we had participated in as well.

CHAMBERLAND: That's good to hear, because I know oftentimes at CDC sometimes there's an inadvertent disconnect between the laboratory and the epidemiologic sides of the picture, but it sounds like from early on that there was a natural joining up. for any type of study that CDC did, be it a case-control study or a cohort, a long-term follow up, they obviously wanted to characterize the T cell counts in these individuals; how many helpers, how many killers or suppressor cells. I can see then that you would be overwhelmed with just supporting our own in-house studies.

NICHOLSON: The other thing is that looking at basic research in human 00:32:00immunology, there was more and more information, or more and more studies showed that you could actually find other kinds of cells, using monoclonal antibodies, that subdivided in a functional way -- the lymphocytes, the T helper cells. There were helper-inducer and helper-suppressor cells, and those reagents to those particular markers became available. It was probably in the late 1980's that we started asking questions about-- are there ways that we can better characterize the immune system, or are there markers for progression that we can identify in these patients. We did a number of studies trying to figure out if we could answer some of those basic immunology questions.

CHAMBERLAND: The field of immunology is evolving at the same time--

NICHOLSON: Exactly.

CHAMBERLAND: -- that the AIDS epidemic is evolving.

NICHOLSON: Yes.

CHAMBERLAND: Do you think that the AIDS epidemic was an impetus? Did it help push immunology along maybe a little bit faster?

NICHOLSON: It probably did. It was a great boom for the technology. The flow cytometers that were developed were largely developed to help with that. Looking at the immune system and how it works was probably very instrumental through the AIDs efforts.

CHAMBERLAND: Let's talk a little bit about some of these immunologic studies that you did and others in the lab did in conjunction with the epidemiologic studies. The initial finding is the shocking lack of T4, T helper cells and an 00:34:00increase in the T8 or suppresser cells that had not been seen before. Again, it's hard to remember there was a time that we didn't actually know this, but there were a number of different of studies that CDC undertook to try and characterize this phenomenon in different populations. At the same time, for example, it wasn't long after full-blown AIDS, as it became known as, the end stage was appreciated, that [CDC received] reports of primarily gay men, but also intravenous drug users, having this unexplained what was called "lymphadenopathy syndrome." They weren't really sick, but they just had a lot of palpable lymph nodes. This was an area that CDC was interested in learning more 00:35:00about. Can you talk a little bit about some of the laboratory studies that tried to characterize what was lymphadenopathy syndrome all about, from an immunologic perspective?

NICHOLSON: It was interesting, because Tom Spira, who worked in our group, was the principal investigator for a study in Atlanta of homosexual men. He enrolled, I don't know how many, probably much more than a hundred, I think, men, in his study. His intent was to follow them longitudinally over time to see how they progressed.

CHAMBERLAND: If they progressed.

NICHOLSON: If they progressed, and that collection of specimens over time was a great treasure trove for us to go back and look at what it was that we could-- if there was anything unique about the immunology of those men. We looked not 00:36:00only at the markers that they had on their cells, there were also studies done on viral load and immune function. It was a great way for looking at the immune system.

CHAMBERLAND: Because people are really wanting to know if there's any way you can "predict" who's going to go on. So, what were you learning at the time?

NICHOLSON: We learned that there were subsets of the T helper cells that may have had some role in prediction. At the end of the day, all of those studies basically showed that the CD4 cell alone was probably the most interesting and predictive of progression.

CHAMBERLAND: Of progression of the disease. As the CD4 counts started getting lower and lower, you had information about their clinical [status], because Tom 00:37:00was collecting data about their clinical illnesses and diseases and also epidemiologic information about behavior-type information, you could correlate all of that.

NICHOLSON: We tried in a lot of different ways to correlate immune function with clinical picture, also with virology, as we knew more and more about the virology.

CHAMBERLAND: Right, because the virus was identified a couple of years later, in 1983. Then there becomes available serologic testing and culture and all of that.

NICHOLSON: Right. Culture, viral load, and--

CHAMBERLAND: Now you're working with another laboratory component of CDC, the virology group?

NICHOLSON: We did work with them as well.

CHAMBERLAND: OK. So, you're all in parallel, multiple streams of research, the epidemiologic, the virologic, and then your immunologic, all trying to put together a composite picture?

NICHOLSON: Right.

CHAMBERLAND: Interesting. At the end of the day, as you said, the decline in the counts of CD4 was the most predictive of someone going on to develop AIDS. There was another-- in preparing for the interview, I looked at a number of the other studies and this was actually, just kind of took my breath away. Before the discovery of the virus and the ability to test for it, CDC was embarking on studies, the classic case control study, trying to identify risk factors as to who would go on to develop the disease. So, comparing at that time gay men who had the disease with so-called controls, men who didn't appear to have the disease, and you went back and tested those control samples, or I'm not sure if it was you personally, but that was one of the laboratory's projects, was to go back and look at some of these controls and found that in reality they weren't 00:39:00really controls, that lots of them were actually infected.

NICHOLSON: That's right.

CHAMBERLAND: What were some of the other immunologic functions that you looked at in these study populations? You talked about functionality. What were some of those studies?

NICHOLSON: We looked at their ability to respond in the test tube to various antigens or just a general stimulus, and they had a decreased ability to do that. That tells us that their immune system isn't functioning the way it should. What would normally happen is that, if those cells were exposed to some substances that would stimulate B cells, you would get an increase of cells in 00:40:00culture or proliferation, if you will, of those cells and culture. Those kinds of studies were in a way rather nonspecific, but in a way specific. Not really specific, but they were general studies that tell you a little bit about whether or not the immune system is functioning well.

CHAMBERLAND: These parallel work streams were all sort of coming together to show a picture of declining CD4 cells, increasing viral load, and antibody titer, if you will. You also looked at whether other cells, cells other than lymphocytes, could be infected, correct?

NICHOLSON: Yes, yes. Because CD4, which was shown to be the entry point, the CD4 00:41:00antigen on the T cells was the entry point for HIV into the cells, we also knew that monocytes--

CHAMBERLAND: Another type of white cell.

NICHOLSON: --another type of white cell had CD4 on its cell surface, though not nearly as abundantly found on the cell surface. We asked the question whether or not monocytes could also be infected by HIV. It was a hard road to go down, but eventually we were able to show that there is some infection of monocytes with HIV, though not to the level as you would see in T cells.

CHAMBERLAND: Does that have any practical clinical significance, if monocytes become infected?

NICHOLSON: I don't think it really had that much of an impact. Monocytes are cells that are involved in fighting infections. They are not the primary cells, 00:42:00but one of the kinds of cells. The neutrophils, who are also involved in just general fighting infections, seemed to be in relatively normal numbers.

CHAMBERLAND: I want to go back to the point you mentioned-- so there's lots of specimens coming in, in relationship to the studies and since these were coming in from the field, from clinical investigators, or other institutions, did questions arise about the validity, if you will, of your testing of lymphocytes, if it was impacted by various storage conditions, and was this area that was important to pursue because I assume if, a lot of times tests are affected by 00:43:00storage conditions, room temperature, refrigeration, frozen. So, what were your findings there?

NICHOLSON: Exactly. It was interesting. I guess early on, it was always assumed that if a laboratory did CD4 cell testing and that same specimen was tested in your lab, that those results would be the same. Well, that wasn't the case. There were a number of issues that had to do with how valid a CD4 cell result was. That included when the specimen was collected, what anticoagulant or what color the tube was that it was collected in, if you would, and how long before it was tested. It wasn't until probably later in the 1980's that the questions as to whether or not the results were valid from lab to lab were asked. There 00:44:00were a number, or at least a couple, of studies that were done, where specimens were shipped around the country and various labs were asked to analyze those and test for the T cell markers to determine whether or not those answers were all the same. It was pretty quickly learned that there were some labs that were pretty good about doing this and other labs that weren't so good.

We embarked on a number of studies just to look at what the optimal conditions were for specimen management and learned that there were some anticoagulants that were good, much better than others, and that you could hold them for a number of days and certain anticoagulants under certain temperature conditions, 00:45:00because temperature had an impact as well. You don't want them heated up, and you don't want them frozen. We did a number of studies looking at those parameters. It's also interesting because monoclonal antibodies varied; monoclonal antibody to a CD4 antigen from one company and another company might not be the same. We found differences in CD4 monoclonal antibodies early on as well, which was interesting; disconcerting in a way, because we learned there were some antibodies that just didn't react with some people's T cells, and we may have been calling people AIDS when they weren't. The interesting reason was 00:46:00we suspected that something was wrong, because when you look at T cells-- the T cells are generally called CD3 cells because that's the number designation for the marker that was on all T cells-- are comprised of primarily CD4 and CD8 cells. When you add those two values from CD4 and CD8 together, for example, 20% and 80%, you should get 100% CD3 cells. If you were getting a missing population that couldn't be explained, that's when we suspected that there must be some antibodies that are not reacting.

CHAMBERLAND: That are on the cell but aren't being picked up.

NICHOLSON: They aren't being picked up by the reagents that we were using. So, we learned a lot of things in the '80s about CD4 cell testing.

CHAMBERLAND: So, did this allow you to make recommendations to people who were out collecting blood specimens to really lay out, this is the specimen we want in this kind of a tube and stored under these types of conditions? Was that a practical outcome of all of that?

NICHOLSON: Yes, it was. In fact, in the late '80s there were standards organizations that were beginning to put out recommendations for what was then called, and is still called, immunophenotyping, which is the testing that is done to identify these CD4 T cells--

CHAMBERLAND: So, the markers on these T cells--

NICHOLSON: The markers on those particular cells. CDC was one of the organizations that also put out guidelines specific for HIV testing, 00:48:00immunophenotyping in HIV infection, that was published in the MMWR in 1993. There were a couple of revisions that came later, as the technology matured and we learned to do it with fewer reagents in a much simpler way. The NIH [National Institutes of Health] also put out recommendations for testing, based in large part from studies we and others had done, looking at what's the optimal way for collecting specimens, sending specimens, holding specimens, etc. There was a boom, if you will, in the late '80s and early '90s about specific ways in which to do this type of testing.

CHAMBERLAND: I'm going to come back to a couple of things that came up in our 00:49:00conversation, but I want to carry this idea of quality control and best practices, laboratory practices. I would imagine that with the boom in immunology related to AIDS and unrelated to AIDS, cancer and whatever, that more and more entities, laboratories, start acquiring this pretty sophisticated machinery, the cell sorter, and the like. Was there any effort to--I know certainly for simple things like serology studies and Western blot testing that came up for testing for HIV, CDC had a big role in training and education of public health laboratories and quality control. Was there a similar effort under way at CDC about trying to maintain some sort of quality control surrounding 00:50:00these studies using cell sorters?

NICHOLSON: The quality control in and of itself, when we wrote guidelines for how to do the test, we wrote into those guidelines ways in which you could quality control your own laboratory. The training, interestingly, the training that was done was generally done not by CDC in this particular area, but it was done by manufacturers or professional organizations that would hold courses and workshops to help the people who were just new to this technology learn how to do it best.

I think maybe one of the biggest things that was helpful in this area was quality assurance cell send-outs, I think it's what they would call it. A blood, 00:51:00generally a lot was divvied up into many different aliquots, and each one was sent to a different laboratory around the country and then the laboratory could test itself. The results would come back, and they would know not only if they were falling within the range of results they should be getting, but there were some laboratories that got remedial assistance from either NIH or--

CHAMBERLAND: They were flunking.

NICHOLSON: They were flunking--or from the manufacturer or from CDC or from whoever was providing those particular cell send-outs. That was a good way for labs to figure out how well they were doing.

CHAMBERLAND: That seems very fundamental because we spent some time talking about the predictive value of decreasing CD4 counts being very predictive of the development of AIDS, and so these are, in a sense, clinical tests that are being provided to individual patients, let alone forming the core of some research studies, so you want to be pretty confident of your result. So, this was something that you played a role in it sounds like, and very interested in this whole--

NICHOLSON: Right. I was involved with the advisory group for the AIDS Clinical Trials Group that NIH had put on, and also for the Canadian quality control assurance group. They were all doing the same thing, trying to ensure that the 00:53:00laboratory results that people were obtaining were right.

CHAMBERLAND: You mentioned NIH a couple of times and I have a couple of NIH-related questions. First of all, what you've just related, I think some people in terms of this early immunologic research, cutting edge research, cutting edge bench research, that CDC was doing, I think some people might be surprised at that, that they might think well that's kind of what NIH does or did, and CDC, their lab work it tends to be much more applied, if you will. I was struck by that, that CDC was a real player, a real leader in this area?

NICHOLSON: I think in the early days we had a lot of access, if you will, 00:54:00through the epidemiologic studies, to specimens, and it provided materials for us to ask some basic questions. I think later, as money became a lot more available to universities and other organizations, if you will, that they got into the game as well. There was a lot of activity in the later '80s and early '90s, and a lot of advances were made in HIV from the funding that was coming in this area.

CHAMBERLAND: So, in these early days, which is the focus of our series, then it is very gratifying to see CDC trail blazing in this area. In those early days, or did it take a while, was there any collaboration with NIH laboratories on some of this immunologic work, T cell studies that you were doing or was it 00:55:00really just coming out of the lab here?

NICHOLSON: There were some colleagues that we were engaged with. I wouldn't say that we working side by side. There were some in the area of immunophenotyping, if you will, at NIH that we did work with, to look at the test itself. The AIDS Clinical Trials Group-- I was involved in that group from the very beginning, I think in large part because when they had established the AIDS clinical trials, they had set up an immunologic and a virologic component at NIH to look at the testing and follow all of that. The man who was involved in that on the 00:56:00immunologic side was [Dr.] Jonathan Kagan. He and I had met actually at a meeting for the, what is now APHL's, [the Association of Public Health Laboratories] concern about T cell testing for HIV infection as well. I did a lot of collaborative work. In fact, some of the work that we had done looking at the quality assurance and the conditions for collection of specimens was done in collaboration with NIH. The basic research, not so much. It was the applied research part that was.

CHAMBERLAND: I also wanted to come back to an important piece of research that came out of the lab here at CDC that you just mentioned very briefly in passing, and this was the series of experiments that CDC undertook under Steve McDougal's 00:57:00leadership that provided insight into how the HIV virus infected T cells because the reason these CD4 cells declined over time is that the AIDS virus basically got into the cell, took it over, and goodbye normal CD4 T helper cells. So, a lot of interest in trying to understand how it is that the virus gets into the CD4 cell. So, can you take us a little bit through some of the ways in which this was approached. I mean this was groundbreaking work to try and establish how the virus came into contact if you will with the CD4 cell.

NICHOLSON: I guess the first challenge was getting the virus.

CHAMBERLAND: Oh, really?

NICHOLSON: I believe the virus that we got came from [Dr.] Luc Montagnier from 00:58:00France. Steve McDougal in our laboratory was the one who decided to take it upon himself to do many of these studies. He squirreled himself away in a little laboratory in the bowels of CDC somewhere in the basement and started culturing the virus. Then [he was] trying to concentrate that virus, because it became apparent that he wasn't going to be very successful without having concentrated it in a way. Then he looked at what it was on the virus, if you could block viral entry by covering those CD4 receptors with antibody so that the virus wouldn't be able to bind. That's how eventually he was able to show that it was the CD4 receptor that in fact was the way that the virus attached to the cell, 00:59:00and of course it has to attach before it can enter the cell.

CHAMBERLAND: And a very specific protein. He identified --

NICHOLSON: Exactly.

CHAMBERLAND: a protein called--

NICHOLSON: --gp120, right.

CHAMBERLAND: glycoprotein number 120, was the very precise--

NICHOLSON: Exactly.

CHAMBERLAND: --point of contact for the virus to--

NICHOLSON: --enter, to hook on to--

CHAMBERLAND: -- for the virus to hook on, if you will to the CD4 cell. And there was a very well-regarded paper that came out in Science, you were a senior author, Steve was first author. This was a big deal wasn't it?

NICHOLSON: It was a very big deal. It was something that people had believed was probably the way it happened, but to have that actually demonstrated was a landmark find.

CHAMBERLAND: Because this was, I think, viewed as the definitive proof that this is how the virus gained entry.

NICHOLSON: Right. Since there were a number of viral proteins, and that being 01:00:00the one that was primarily responsible for attachment, then therapeutics could be designed.

CHAMBERLAND: So, that's the motivation? Can we find, yeah can we find a way--

NICHOLSON: Right. But we don't do the therapeutic side. iI provides information for those people who do.

CHAMBERLAND: The basic research that would hopefully lead on. I don't think in the end, this paid off maybe as well as people might've expected but it gave people promise and hope that ok if we can block this. So, now you've told us how the virus hooks on to the cell, it's up to others, pharmacy industry and others to try and figure out a way to block this. It was published as I said in Science. I read somewhere that this particular paper in Science was cited more than 120 times in the first 18 months of its publication. It also won the CDC 01:01:00Charles Shepard Award which is presented annually to the best manuscript on original research that's published by a CDC scientist or scientific group. What was your reaction, your colleagues' reaction to this? Did you really appreciate the enormity of what this was all about in your work or did it take you time to process that over the years?

NICHOLSON: We knew it was important at the very beginning, but we knew there was a long road ahead to take advantage of the finding in fighting the AIDS epidemic. So, yes and no.

CHAMBERLAND: I saw a sketch, the Charles Shepard Award, one of the CDC artists 01:02:00[Ed Biel] would always draw a sketch of the award recipients, and apart from Steve everyone on the poster is a woman.

NICHOLSON: Yes. Steve and his harem.

CHAMBERLAND: Is that what you became known as? You mentioned that it was a pretty small group when you first started. Obviously, it expanded, a lot of women. Who were some of the people that you worked with apart from Steve, as time marched on?

NICHOLSON: Steve and Tom Spira, as I mentioned.

CHAMBERLAND: Yes. Tom was an immunologist and a physician immunologist.

NICHOLSON: Yes. Yes. There were a lot of them. Bonnie Jones and Marjorie Hubbard were from the old immunology group and were particularly valuable in doing the immunophenotyping studies. I worked with them very closely. Carol Aloisio came 01:03:00on. [Dr.] Linda Martin was another investigator; she actually looked at the inactivation of HIV. She did a lot of the studies looking at what it took to do that.

CHAMBERLAND: Ultimately, I think it was heat.

NICHOLSON: It was heat, yes, looking at the heat inactivation.

CHAMBERLAND: In terms of --to look at the inactivation of virus in these hemophiliac factor VIII --

NICHOLSON: We actually had looked at fixatives that would kill HIV in the preparation of the specimen. But some of the other people on the virology side were Susan Kennedy, obviously, and Donna Sasso was involved. There were a number early on. David Kickliter, actually, David Kickliter, who didn't stay much longer after I had come on, was trained to run the cell sorter, and he never 01:04:00ended up running the cell sorter. David Cross worked for me, and Caroline Dawson did some of the flow cytometry studies as well. [Dr.] Janine Jason, who was in the hematology group, came over to our group. I think that was probably about 1990 or so.

CHAMBERLAND: She had some immunology training.

NICHOLSON: She had some immunology training; she was a pediatrician.

CHAMBERLAND: When I look at the lab, your lab cranked out in a relatively short period of time in the mid- to late-'80s--again we're talking about the early response, the work clearly went past that time. But there was a lot of papers that came out, a lot of research that came out, and at the beginning of the interview you mentioned you were a new mom when you first started. So, this is 01:05:00obviously happening at a time when there's a lot going on in your life apart from work. What was it like in the laboratory environment then to keep up with the pace? Were you exhilarated, exhausted? I'm trying to get a sense of what that must've been like.

NICHOLSON: For me it was satisfying. It was a lot of work but a lot of opportunities. A lot of opportunities, because we had funding. We finally had funding to do some of this work. We had the specimens to do it with, and we had all kinds of ideas that came up. It was a very satisfying time of my career. A lot of collaborations, you're right. We did do a lot of collaborations, both within CDC as well as outside of CDC.

CHAMBERLAND: Not being a laboratorian, I've always been curious about how laboratorians design their studies, their research. So, again, when AIDS comes along, this is unchartered territory. Immunology is evolving, your tools are evolving, your technology, your machines, the computers. So, would you sit around and banter among yourselves, like "Okay, here's a question we want to answer; what do we think is the best way to go about approaching this?" Or is it more that you sort of individually would think this through. I'm just very curious how--

NICHOLSON: I think it's a combination of both. We had lab meetings. Novel idea. But we had lab meetings on Tuesday mornings when we all gathered in the hallway. 01:07:00Steve put a chalkboard up in the hallway, and he would bring Dunkin' Donuts. We would bring our lab chairs and line them up and down the hallway and sit, and he would talk, or we would talk, have presentations. One of the things we talked about was interesting questions that we think we could answer, or questions that should be answered. There was a lot of banter in those forums. Also, interestingly, we would go to the cafeteria en masse, it seemed like, sometimes for a coffee break and for an afternoon break and for lunch. I guess people in the cafeteria must have thought we were just all together all the time.

CHAMBERLAND: Joined at the hip.

NICHOLSON: Joined at the hip. But we got a lot of good ideas there as well.

CHAMBERLAND: That's interesting because I think so often people are accused of working in silos or the intensity of the work often means that people don't take time to have a little bit of down time, a little bit of socialization. So, it sounds maybe some of the more creative ideas came about in some of these more relaxed, if you will, environments of cafeteria--

NICHOLSON: I think so. Then not only that, but if you had read a paper and think that an extension of that or another question, or a different way they could have asked a question, or whatever, and then to have that conversation in the group. We were a very congenial group. I think everyone pretty much got along with everybody else, and we socialized together some. It was just a good 01:09:00atmosphere in which to be working.

CHAMBERLAND: Well, sadly your chief Steve McDougal passed away some three years ago and his stories cannot be captured in this oral history project. And you worked with Steve for a long time. What can you tell us about Steve?

NICHOLSON: He was a brilliant scientist. I was astounded at the very beginning. He took me under his wing, I told you, as a graduate student finishing up my thesis. One of the things that just impressed me about him was, he had a manuscript that had been returned to him. He had submitted a manuscript to the Journal of Immunology for publication and had the manuscript returned to him with nothing but comments on it and [it was] published immediately. This was unheard of. Usually, you had a reviewer that wanted you to make revisions. No revisions were needed. That I don't think was the only publication that he had 01:10:00no revisions requested for. He was just such a brilliant guy and funny. He had a big personality.

CHAMBERLAND: You mentioned he was your Branch Chief, I guess ultimately. I know you've gotten reorganized a few times. But he was a Branch Chief, but he did bench work.

NICHOLSON: He did.

CHAMBERLAND: Is that unusual?

NICHOLSON: I think it was unusual. He was an MD. He wasn't a Ph.D. He did a lot of bench work, and his office was the bench. We were in a hallway where there were two offices. Tom Spira had one office, and Steve had an office at the other end for a short period of time. He gave up his office and gave it to me. So I was in the office at the beginning of the hallway. Steve's office was the lab.

CHAMBERLAND: Really? You had to go into the lab to find Steve.

NICHOLSON: You had to find him in the lab, and he was among our-- the pieces of souvenirs that were part of our souvenir contest, "best souvenir."

CHAMBERLAND: From where? When you took a trip somewhere?

NICHOLSON: Anywhere you took a trip, you came home with a souvenir. Whoever came home with the best souvenir--we had some very interesting souvenirs-- but they lined the shelf above Steve's office at the bench. It was a fun group.

CHAMBERLAND: It sounds like it was a really good work environment.

NICHOLSON: It was tacky souvenir, not best. Tacky souvenir, best tacky souvenir.

CHAMBERLAND: Looking back, are there any aspects of CDC's laboratory response to 01:12:00the epidemic that you think we fell short or could've done a better job? And conversely, where did we excel?

NICHOLSON: Oh, that's a good question. I think we excelled at getting information out. I think that was probably very helpful. I think in the virology area there were many places where they excelled. The use of PCR [polymerase chain reaction], the introduction of PCR for testing in HIV, was a huge step forward.

CHAMBERLAND: That was the ability to test for antibodies for the virus.

NICHOLSON: That was the virus. That was testing for the virus itself, because then it went on to doing quantitative viral load.

CHAMBERLAND: Because previously it had just been culture, which was very tedious and laborious.

NICHOLSON: Exactly. It was very tedious. I think that [PCR] was great. There was a lot of good research applied, and some basic, mostly applied research that came out of CDC that was-- particularly in the early days, that helped guide future investigators in lines of research as well. I don't know if there were any particular shortcomings that I can think of.

CHAMBERLAND: So, this occurred-- you're working in the early laboratory response to AIDS in your field. This occurred at a very early point in your career at CDC and also at a point in time when you were just beginning to raise a young 01:14:00family. Over the years, have you had an opportunity to reflect on how this period of time in the work that you did influenced you both personally and professionally? AIDS-- there's been obviously new pathogens down the road, but AIDS was a special deal.

NICHOLSON: AIDS was very special. It's interesting. When I worked in Galveston, I worked among a group of scientists that were all focused on the same problem, but they all came at it from different ways. It was a team effort. We had about 20 people or so in our team. I liked that environment, and that was the same environment that was created at CDC. We had a team, if you will, and we're all doing something a little bit different, but it all came together to ask and 01:15:00approach a question of interest. I've always appreciated that kind of atmosphere. It was instrumental probably in me moving to other areas at CDC, because I was no longer necessarily a face that nobody saw anywhere or didn't know anything about. I probably became a little bit more visible to folks at CDC than I would have otherwise.

CDC has been a great career for me. I couldn't have asked for a better career. I think it's because of the idea that we work in public health. That was something I never dreamt about doing when I was growing up. I wanted to be a marine biologist or a mathematician or something else. To end up in public health was 01:16:00probably the best thing that could have happened. It has such a humanitarian aspect to it, is what I really learned to appreciate.

CHAMBERLAND: Any closing thoughts, anything that we haven't covered?

NICHOLSON: Oh, probably not.

CHAMBERLAND: Well that's a good place to end. Thanks so much, Jan. Thank you for sharing your memories and your thoughts with us.

NICHOLSON: Sure. It was fun.