ABSTRACT

First Person is a series of interviews with the first authors of a selection of papers published in Journal of Cell Science, helping early-career researchers promote themselves alongside their papers. Lotte de Winde is the first author on ‘C-type lectin-like receptor 2 (CLEC-2)-dependent dendritic cell migration is controlled by tetraspanin CD37’, published in Journal of Cell Science. Lotte conducted the research in this article while a PhD student at the Radboud Institute for Molecular Life Sciences, The Netherlands, under the supervision of Prof. Annemiek van Spriel and Prof. Carl Figdor. She is now a Postdoctoral Research Fellow in the lab of Dr Sophie Acton at the MRC Laboratory for Molecular Cell Biology, UK. Here, she studies the plasma membrane organisation and dynamics of podoplanin and its partner proteins on fibroblastic reticular cells (FRCs), and how changes in this organisation shape downstream signalling and FRC function.

Lotte de Winde

How would you explain the main findings of your paper in lay terms?

Dendritic cells (DCs) play an important role in activating an immune response. They take up an unknown, foreign particle, that is, from a bacteria or virus, and bring this to the lymph node to show it to other immune cells, T and B cells. Activated T and B cells will then attack and work to remove the bacteria or virus, and any infected cells. When DCs take up a foreign substance, they become activated and begin to travel to lymph nodes. One of the changes that allows DCs to move is the expression of C-type lectin receptor CLEC-2 on their surface; CLEC-2 interacts with another protein, podoplanin, on a cell type called lymphoid stromal cells during DC migration. It was unknown how the CLEC-2–podoplanin interaction was controlled. Our data showed that another type of protein, a tetraspanin named CD37, sits next to CLEC-2 and helps DCs to migrate on lymphoid stromal cells. CD37 controls both CLEC-2 recruitment on the DC surface when binding to podoplanin and also how the lymphoid stromal cells respond to the DCs crawling over them. The latter is an important first step in lymph node swelling.

“This project has taught me that you cannot do science alone, that you need to collaborate and take every opportunity to learn and bring your ideas and scientific story to a higher level.”

Were there any specific challenges associated with this project? If so, how did you overcome them?

This project started off as a side-project in the first year of my PhD, because my main project was on hold for a few months. At that time, we only had the co-immunoprecipitation data from our collaborator Dr Mike Tomlinson and his PhD student Alexandra Matthews, showing a specific interaction between CLEC-2 and CD37. I did some initial experiments looking at CLEC-2 expression and cytokine production in Cd37-deficient immune cells. After a few months, I could continue with my main project, which left this project on hold for two years. However, I really liked this project and had a lot of ideas for experiments, but I didn't have the tools or experience to do it. I discussed this with my PhD supervisor and she gave me the opportunity to apply for travel grants to visit and do some experiments in the lab of a collaborator, Prof. Steve Watson in Birmingham, UK. In the final year of my PhD, I went there, which was a challenge in itself as it felt like I had to start from scratch in a new environment. But after the first week, with a lot of help from Steve Watson, I made connections with Dr Helen McGettrick (Birmingham) and Dr Sophie Acton (London) and had a research plan for the next 2 months. The results of these experiments all proved my hypotheses, and after those 2 months, I had two figures to go into a manuscript. The last challenge of this project was in the final stages, where I was finishing my PhD and moved to London for a postdoc, but with the help of my colleagues and collaborators, our story was finished and is now published. This project has taught me that you cannot do science alone, that you need to collaborate and take every opportunity to learn and bring your ideas and scientific story to a higher level.

CLEC-2+ dendritic cells (DCs; smaller black cells) interacting with podoplanin-expressing fibroblastic reticular cells (FRCs; magenta) havestrong phenotypic effects on both cell types. DCs use FRCs to migrate along, thereby increasing their cell area. At the same time, FRCs will lose actomyosin contractility, which is visualised by loss of actin stress fibres, stained with phalloidin (black), and presence of lamellipodia. Cell nuclei are stained with DAPI (blue).

CLEC-2+ dendritic cells (DCs; smaller black cells) interacting with podoplanin-expressing fibroblastic reticular cells (FRCs; magenta) havestrong phenotypic effects on both cell types. DCs use FRCs to migrate along, thereby increasing their cell area. At the same time, FRCs will lose actomyosin contractility, which is visualised by loss of actin stress fibres, stained with phalloidin (black), and presence of lamellipodia. Cell nuclei are stained with DAPI (blue).

When doing the research, did you have a particular result or ‘eureka’ moment that has stuck with you?

I was very happy with the results I obtained during my lab visits in Birmingham and in London. They proved my hypotheses and from that moment I could work towards a manuscript.

Why did you choose Journal of Cell Science for your paper?

Despite studying an immunological process, we decided that we wanted to publish this in a cell biology journal, because this study really focuses on molecular and cell biological mechanisms. I wanted to publish in Journal of Cell Science, as it publishes high-quality research with a clear message and the scientific articles are broadly accessible to a wide range of scientists across many fields of research.

Have you had any significant mentors who have helped you beyond supervision in the lab?

My PhD supervisor Annemiek van Spriel has been and still is a great mentor. She taught me to think independently and take ownership of my projects, which helped me specifically in finishing this project. Steve Watson has been a great mentor in this project. He not only hosted me in his lab, but also brought me into contact with Helen McGettrick, so I could do the experiments I came to Birmingham for. Furthermore, he set up a meeting with Sophie Acton in London during my first week in his lab. For me, this had an unexpected outcome, as it resulted not only in a fruitful collaboration for this project, but is also the lab where I am now doing my first postdoc, and she is now also an important mentor for me. If I look back on my scientific career so far, I've always had great supervisors and mentors along the way who have helped me onto the next step(s), and I'm really thankful to them.

What motivated you to pursue a career in science, and what have been the most interesting moments on the path that led you to where you are now?

I always wanted to become a medical doctor, but I didn't make it through the lottery to study medicine in The Netherlands. I decided to study molecular life sciences at Maastricht University, The Netherlands, and during my second year I realised I really enjoyed solving the molecular puzzle behind a disease. The final decision for a career as a scientist was during a 10-week internship at the end of my Bachelor's – I actually enjoyed doing science in a lab, and I still do!

Who are your role models in science? Why?

My role models are scientists who pursue their ideas with a lot of passion, have fun doing science and help others to improve their research and guide them in their careers.

What's next for you?

My NWO Rubicon fellowship runs until May 2019, but I hope to stay in Dr Sophie Acton's group until 2020, so I have enough time to finish and publish my current projects. Simultaneously, I'm preparing to apply for new postdoctoral fellowships to continue my own research plans and ideas. And if the next years are successful scientifically, I hope to start of my own research group one day.

Tell us something interesting about yourself that wouldn't be on your CV

Every year, together with friends, I volunteer to be a staff member on a children's summer camp. We spent one week camping in the woods in primitive conditions and the days are filled with a lot of sports and games. It is great to be fully away from the rest of the world (i.e. no emails!), and I always return fully reloaded.

Lotte de Winde's contact details: MRC Laboratory for Molecular Cell Biology, University College London, Gower Street, London WC1E 6BT, UK.

E-mail: l.winde@ucl.ac.uk

Reference

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C-type lectin-like receptor 2 (CLEC-2)-dependent dendritic cell migration is controlled by tetraspanin CD37
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J. Cell Sci.
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jcs214551
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