I asked Laurence Pearce to write a little about his time at the Cardiff CLL group. This is what he has written. Laurence worked hard and it was great to have him in the team. He is now working in the NHS in Nottingham. Good luck, Laurence.
My experience as a PhD student with the Cardiff CLL group.
In the spring of 2008 I moved to Cardiff to carry out a PhD under the supervision of three highly motivated scientists (Drs Brennan, Pepper and Fegan). Their aim was to dissect the biological mechanisms underpinning the development and progression of chronic lymphocytic leukaemia. The group had developed a particular interest in elucidating the role of the CD38 glycoprotein, whose presence on the surface of the CLL B-cell results in an unfavourable outcome in this disease.
My initial aim was to develop a technique to induce the expression of CD38 on the surface of CLL cells which primarily did not express the glycoprotein. This would provide me with the opportunity to compare cells from the same source with the only difference being the presence of the CD38 glycoprotein on the surface. Any changes in the cell’s behaviour could then be attributed to this molecule.
After investigating three different techniques I developed a highly successful method of expressing CD38 on the surface of the CLL cells using a HIV-1 derived lentiviral delivery vector. The effectiveness of this cutting edge genetic engineering technique led to the publication of a manuscript describing the work that I had carried out in the first year of my PhD.
I proceeded to carry out a range of investigations to observe the genetic and cellular changes following the induced expression of CD38 on the surface of the CLL cells. These included microarray, QRT-PCR, multi-colour flow cytometry, calcium flux assays and confocal microscopy. As a means of stimulating CD38, I introduced the CD31 molecule (the only described non-substrate ligand for CD38) in the form of a soluble antigen or bound to the surface of adherent helper fibroblast cells. Carrying out these investigations on multiple patient samples resulted in the accumulation of a great deal of data from which to generate high quality figures for my thesis.
The final stage of the project involved writing up the work and with the help of my supervisors (and copious amounts of tea) I managed to complete the thesis and hand in. The viva examination date was arranged and following a short period of intense revision and background reading I successfully defended my thesis to the panel of examiners.
I thoroughly enjoyed carrying out my PhD with the Cardiff CLL group and made a lot of great friends who I will continue to discuss science, rugby, music and many other topics with, probably over a beer in the local pub.