One of the stand-out papers in the archives of Immunology, one of the BSI's official journals, is a 1970 work by Martin C. Raff entitled ‘Two distinct populations of peripheral lymphocytes in mice distinguishable by immunofluorescence’.1
Martin Raff worked at University College London (UCL) for over 30 years until his retirement in 2002. He is now an Emeritus Professor at UCL, and continues to contribute to the scientific community through his work on scientific advisory boards. He trained as a clinical neurologist in Boston before pursuing an opportunity to improve his knowledge of basic science in Mill Hill, London. The move brought him to the heart of the UK's immunology community, where he worked with remarkable scientists such as Avrion Mitchison, and produced a series of papers investigating lymphocyte biology.
The remarkable paper he published in Immunology outlines Raff’s investigation into the heterogeneity of lymphocytes, then an important and relatively new field of study, and how to distinguish classes of lymphocyte. To investigate the question, Raff compared lymphocytes from normal mice and from T-cell-depleted mice, discovering that one class of lymphocyte (now known as B cells) were Ig-positive and theta (θ)-negative, while T cells were θ-positive and Ig-negative. He had shown earlier in a Nature paper that Thy-1 is a marker for mouse T cells, which enabled this field of enquiry to flourish.
Nearly 40 years after this remarkable paper was first published, Immunology’s Editor-in-Chief, Danny Altmann, braved a rainy November evening to meet Martin in his office at UCL’s MRC Laboratory for Molecular Cell Biology and talk more about T cells and B cells, a remarkable four years in the history of immunology, and the enduring importance of a good title.
Danny Altmann: It is wonderful to meet you Martin. I wondered if you could tell me more about the story behind this paper?
Martin Raff: I’d only been doing basic science and immunology for about a year and a half when I sent this paper off to the Journal of Experimental Medicine, which was quite unusual! In my view it was an important finding, but I was new and I just didn’t know that much about how you deal with the politics of science. Anyway, it was bounced right back without refereeing, saying that there is nothing novel here. I have to say, in re-reading it now, the importance of the finding was certainly not obvious from the title. It’s a pathetic title! Even in the discussion the real importance was not emphasised. The quick rejection should have pointed out to me that this was not the right title.
Titles have always been important. You know that 0.0001% of people are going to read your whole paper. 0.001% will read the abstract. And 0.01% will read the title. So, 99.9% of your readers will miss the message if it is not in the title.
Anyway, I was unsure what to do next so I went to my mentor, Avrion (Av) Mitchison, to ask for advice. He was on the editorial board of Immunology, so he suggested I submit it there.
DA: Av had the idea that the thymus was something to do with the different kinds of lymphocytes or programming subsets of lymphocytes. So where did it start from?
MR: I was a board-certified neurologist when I first started in science at Mill Hill in London. I had decided to do a few years of basic science to gain some lab experience. I chose Av’s lab because I couldn’t understand what he wrote about – I thought maybe others would struggle too and be put off applying to work with him, so I’d have a better chance of getting in his lab.
When I arrived he gave me a project. He said he’d heard Arnold Reif describe this antigen called θ (now called Thy-1) which was on thymus and brain cells, but not in peripheral lymphoid tissues. Av thought, ‘well, T cells come from the thymus, maybe this would be a good marker for them’. This was the early days of T cells and B cells, and one was really desperate for markers to distinguish them.
So that was my project, and it worked even though I knew no immunology. That was largely because I was surrounded by many immunologists who knew a hell of a lot, and I just kept asking questions and asking how to do things. My first paper was published in Nature and showed that Thy-1 marked T cells. It made use of mice that were depleted of T cells, and I found that the mice had dramatically reduced numbers of Thy-1+ cells in their peripheral lymphoid organs, so it was pretty clear that Thy-1 was on T cells. And then I wanted to use immunofluorescence to look at Thy-1 on the surface of T cells. In a control experiment, I found that fluorescent antibodies against mouse immunoglobulin (Ig) on its own stained mouse peripheral lymphocytes; and Roger Taylor, working across the hall, found the same thing using radiolabelled anti-Ig antibodies and autoradiography. We published our findings together, showing that there is Ig on mouse lymphocytes, but we didn’t know which type of lymphocytes they were.
The Immunology paper showed that the lymphocytes with Ig on their surface did not have Thy-1 on their surface, suggesting that they were B cells. The finding that the Ig-positive cells were increased in T-cell-depleted mice, whereas Thy-1-positive cells decreased, supported this conclusion.
DA: It’s good science.
MR: It’s good science, but that should have been the message in the title of the paper!
DA: That’s the joy of hindsight, isn’t it?
MR: Well, it should have been in my head, because when I sent my first paper to Nature, John Tooze, the handling editor, said ‘really interesting stuff, but you’ve got to change the title’, which should be the main message of the paper – that θ (now Thy-1) can serve was a T cell marker in mice. I can’t remember what the original title was – but it was clearly as boring as this one on the Immunology paper!
What’s interesting about that was that Schlesinger and Yron published pretty much the exact same findings about θ in Science a month before I did and reached the same conclusion. But their title was as bad as my original one, and they never received as much credit as I did, thanks to John Tooze.
DA: And Jacques Miller was onto something similar as well?
MR: Not quite. In fact, when Miller came by to visit Mitchison back then, I’d only been in science for about 6 months. Because he was a competitor of Av’s, I asked Av what I should do. Should I tell him what I was doing? Av said, ‘yes, tell him absolutely everything.’ So, I did, and, when Miller asked if he could have some anti- θ serum. Av said, ‘give him as much as you can spare’. This was Av’s way, and it turned out to be an invaluable lesson for me that served me well for the rest of my career.
DA: What were your impressions of UK immunology? What kind of crowd were they?
MR: Well, I was new to the field so couldn’t compare it with anything, but it was all terribly exciting. I mean, Mill Hill was abuzz with great immunology. The BSI was really important. That’s where I learned a lot of immunology.
DA: How were BSI meetings in the ‘70s? What was exciting to see?
MR: There were hundreds of people at the BSI meetings. They were big meetings, because immunology in Britain was big. The BSI meetings had people from all over Europe and America. They were very good meetings.
It was exciting to see the beginning of the T and B cell story. Tony Davis was one of the people working on T and B cells, Dick Gershon was doing research with Tony, and Av had attracted a lot of outstanding visiting immunologists studying T and B cell interactions – it was the beginnings of understanding these interactions. That’s why Av was in competition with Mitchell and Miller, because they were looking at these interactions in one way, and Av was doing it another way, but they were coming to similar conclusions.
For me, starting from scratch and knowing almost nothing about anything, let alone immunology and science, those three years were the most important and productive in my career. I mean, it was downhill from there!
A point that I make over and over again whenever I can is that Av would not put his name on any of my papers. That’s a remarkable thing. Av once told me, ‘I think when you’re a mentor you have an obligation to your students to promote their careers. And clearly, letting you publish on your own greatly promoted your career.’ And yes, it did. But on the other hand, the θ project originated with him, and he made the first anti-θ antiserum for me. It’s not right that his name was not on at least the first paper – it’s an inaccurate history of what actually happened.
DA: And he was a young man, it wasn’t like his career was safely established at that point.
MR: Av did all of his own experiments, and he mainly published his own results. He didn’t rely on his students’ and postdocs’ experiments. He wasn’t treating me differently from the other postdocs and students. Anyway, it’s why I’m a scientist today. I owe him big time. He is a great scientist, mentor, and person.
DA: He was very influential in my career as well. I think to young immunologists who inhabit a world of 100,000 lymphocyte subsets and mass spectrometry, it’s very hard to understand a time of only one or two lymphocyte subsets.
MR: It is so interesting to compare then with now. Those times were unusual. In part, because so little was known that almost every set of experiments I did was a discovery. The immune system is so accessible. I mean, you take the thymus out of a mouse and you get a hundred million individual cells. It’s remarkable. When I started to do neurobiology and was able to get tens of thousands of individual cells out of the optic nerve of a rat, I realised what an exception the immune system is: In some cases, I was able to do one experiment in the morning and another in the afternoon – then read the results and plan what I’d do the next day. Those were special times.
One the other hand, the tools were relatively primitive. For example, you had to make your own antibodies and label them; there were no commercial sources or kits. But the life of a scientist was relatively simple, I think. There was relatively little career anxiety: I was not aware of anyone worrying about a job, or about getting papers published, or getting grants. There were so few scientists compared with today.
DA: Lots of people who might not know you were an immunologist will know your textbook, Molecular Biology of the Cell. Decades of scientists have grown up with it. What do you think about students now who perhaps get more of their information from Wikipedia than from a book like yours? Do you think they’re missing out?
MR: That’s a very important question. As you know, the textbook publishing industry is in some disarray – partly because of the rental market and ebooks, but also because fewer students read textbooks, especially in the US. There is the added problem of ebook piracy, especially in China.
When we started writing Molecular Biology of the Cell, Bruce Alberts, Keith Roberts, and I spent a summer at Jim Watson’s house in Martha’s Vineyard. None of us apart from Jim had ever written a textbook, none of us had ever taught cell biology. We set out to emphasise concepts, trying not to load the student with facts unless there was a story to tell. As so few stories were known, we often had to make informed guesses to give the facts a plausible context. I think this helped make the book special.
Watson was a co-author on the first few editions. He already had another a very successful book, The Molecular Biology of the Gene. He really brought the idea of conceptual writing from his first book, and we carried it over into this book. The first two chapters that were written (one of them was in immunology) went out for review to university teachers and got trashed because the reviewers thought this conceptual approach would not work for an undergraduate textbook. Nonetheless, the publisher was bold, and with Watson as a famous and successful author, he borrowed a million dollars and went ahead anyway.
Are textbooks still needed in the age of Wikipedia? I think probably yes. If a student wants to find out about a cell biological subject using Wikipedia, it could be pretty tough going. The motivation and storyline are usually missing: ‘why should I be interested in this subject?’ Another reason I think textbooks will remain useful is that the amount known about most subjects is now probably hundreds of times greater than what it was when I was an undergraduate. And the brain has not increased significantly in its ability to learn. When we wrote Molecular Biology of the Cell there were very few interesting stories you could tell about a particular protein, whereas now there are many hundreds of stories for many proteins. How can you guide the student through this morass when there are well over 20,000 distinct proteins? Both the teachers and students need guidance, and a good textbook can help provide it, and, importantly, can also provide motivation for why it is worth learning the stuff, although this should be a large part of the teacher’s job.
DA: After your time working on the immune system, you moved into the field of neurobiology. If you were heading the Gates Foundation equivalent for neurobiology, what would you fund?
MR: I would probably invest in new technologies. Paul Allen, who co-founded Microsoft with Gates, set up the Allen Institute, which did a tremendous service for neuroscience by mapping the expression of most mammalian genes in the mouse brain, and, later, the human brain, which greatly accelerated our understanding of these genes in the brain. Although new technologies have already revolutionised neurobiology, given the brain’s complexity, technology is still limiting progress in understanding brain function. That is the safe answer to your question, as trying to predict the future is a mug’s game.
DA: How easy do you think it is now to have a career like yours, which has been so interdisciplinary, and has spanned so many different fields?
MR: It’s a good question. It is certainly much harder now then it was in my day. Just to keep up with progress in one narrow field is now much harder, as so much more is known and so many more scientists are now in the field. It’s great that so much of the information is readily available on your computer screen and that the tools one has to work with are so much more powerful, but thinking time is becoming limiting, which is unfortunate, as an hour of thought can save you months of work.
DA: I think we can be very technologically driven now, and not always in a positive way as we accumulate huge datasets without the brain wattage to analyse them.
MR: True. But, on the other hand, big data can be extremely important: cracking the human genome, for example, revolutionised human biology and human disease research. But big science and huge data sets don’t bypass the need for thoughtful, simple, experiments.
DA: Martin, thank you for your time. It was been wonderful to speak with you.
MR: I should say one other thing about my Immunology paper. When it was published in Immunology, the journal’s impact factor was low, especially compared to The Journal of Experimental Medicine, which was at least ten times higher, maybe more. Yet the paper became my first and only Citation Classic whereas my Nature papers from the same time did not, even though Nature had a much higher impact factor even than The Journal of Experimental Medicine. The lesson I have taken from that is that it doesn’t matter where you publish something: if it is important, it will be found, and that was well before papers were available on the worldwide web, so It is even more true today. So, hats off to Immunology.
This interview is part of 'Celebrating Immunology at 60'. Find out more here!
Immunology has been supporting the work of the BSI since 1958. Profits derived from the sale of the journal are invested back into the BSI, providing major financial support for the Society's activities – including our travel grants, Regional and Affinity group meetings, BSI Congress and more. Find out more about submitting to Immunology here: Call for Submissions.
1 Raff, MC. Two distinct populations of peripheral lymphocytes in mice distinguishable by immunofluorescence. Immunology. 1970 Oct; 19(4): 637–650.
2 A Citation Classic is a highly cited publication as identified by the Science Citation Index, the Social Sciences Citation Index, or the Arts & Humanities Citation Index. They were gathered by Current Contents between 1977 and 1993, accompanied by a commentary by the authors. The full collection can be found here.