Graft-versus-host disease (GVHD). The heart of my supervisor’s research are these four words, much like a riddle in its entirety, it would be better to say the heart of this research relies in solving this particular riddle. However this puzzle goes beyond the typical, with its involvement of tissue culturing, fluorescent microscopy, something rather brilliant known as FACS, and much much more. I felt like one of the Doctor’s companions, journeying on a mission, but instead of bow ties and a fez, I had lime gloves and a lab coat, and instead of a Tardis, we had a centrifuge- not nearly as big but it does consider time and relative dimensions (just not in space). The beginning of this mission involves understanding what GVHD actually is; the scenario is as so: bone marrow transplantation is an upheld treatment for curing some blood cancers, but use of this treatment is limited by GVHD. This is when the donor’s T cells attack not only the recipient’s mutated cells, but also their healthy cells. My supervisor Clare Bennett, and her team have a particular focus, which are immune cells known as dendritic cells. Dendritic cells have a vital role in activating donor T cells to attack patient’s cells.
How? This one question brings about a collaboration of procedures, advanced technology and talented scientists, which I had the honour to work with. Amongst the many techniques I had a chance to witness, the most detailed of which was the tissue culturing of tumour cells. A minor fraction of which entailed using several pipettes (much fancier than the plastic ones in school) to get rid of the tumour cells’ waste, feed the tumour cells the necessary nutrients in order for them to grow and incubating the tumour cells. One of the highlights of tissue culturing for me was using a centrifuge; up to that point the existence of a centrifuge was only known to me through biology classes and drawings in textbooks. So to see this in actuality was like seeing a real life unicorn, but only better. See, a centrifuge is a superb machine, which applies centrifugal force to its content; it takes into account the different densities of the substances that are contained within this fluid, and separates them into a pellet and a supernatant each time the fluid is spun – things, which a unicorn most certainly cannot do. And what a great coincidence that the five marker in my biology exam was on this particular machine…
Now onto this brilliance known as FACS- fluorescence activated cell sorting is a specialised type of flow cytometry. It specifies a method by which a mixture of biological cells can be separated into different containers based upon the fluorescent features and specific light scattering of each cell, which is determined by the markers and the proteins the cell contains, and how abundant it is in each particular cell. As part of my tissue culture experiment, I analysed B16.F10 cells (a melanoma tumour cell line). I put tumour cells in six different mediums, the first of which was a negative control. The other five contained a media known as CASM; CASM contains a cytokine known as interferon gamma (IFN-g), which stimulates the tumour cells to express components known as MHC (major histocompatibility complex [class II]), which they normally wouldn’t do in order to avoid the immune system. My association with FACS featured tagging MHC class II with fluorescent dyes, and processing each of the six samples of tumour cells and seeing which samples expressed the components MHC class II. The results are as follow:
Beyond the tissue culture, the FACS and even the awesome portal door that you have to spin clockwise in, in order to reach the dark room to carry out fluorescent microscopy, the best part of this work placement has to be the people I got to work with. Despite being involved with research that is fundamental to an inevitable breakthrough in cancer research in the upcoming future, everyone welcomed me with open arms. There was Clare who warmly received my presence into her lab, Alastair who helped me carry out the entire tissue culture process, Anjum and Sophie who offered invaluable advice on applying to medical school and Anna and Lia who openly shared their research with me, and were always there to share great conversations with. The research, technology and experiments I witness were undeniably fascinating. However the people that make this happen everyday are even better; they have further fuelled my aspiration to becoming a medic, as well as inciting a spark in me to pursue cancer research alongside.