Tuesday, August 5, 2008

Week Eight

It's really strange, but somehow eight weeks have gone by. I can't believe I've been here for two months now! My constant adventures around the Boston/Cambridge area are going to come to an end...Here is a picture of the Charles. What a beautiful place!



Ellen has finished her time here last week, and Josh, Vanetta, Joe, and Megan have all been getting ready to head off for an annual conference in Australia at the end of this week. I helped Josh work on his presentation and slideshow that he'll be presenting next week, and later shadowed Carrie as she showed me how to do DNA extraction. We started with mRNA, made cDNA, then converted back to RNA and then DNA once again. Tuesday, we ran the PCR for this, and irradiated the mice for Wednesday's bone marrow transplant.

Journal Club was led by Josh this week, and I spent some time really trying to understand the article before the meeting. Surprisingly, I found that it was pretty straightforward, and that I really was understanding most of the data just reading it through myself. The article dealt with an immunostimulatory T cell Ig mucin (Tim)-1 specific antibody that deprograms Tregs. T-regs are a type of CD4+ helper T-cells that are resonsible for immunosuppression and anti-inflammation, which make them key players in tolerance studies as they do their best to prevent a graft-vs-host immune response. With the T-regs "deprogrammed" - or not completely functional in the presence of this 3B3 anti-Tim-1 mAb, transplant tolerance is not possible. I ended up presenting a couple of the figures, which was an initially uncomfortable but altogether rewarding experience. It's really hard to sit in front of PhDs and MDs and attempt explain a scientific figure, but I'm glad I did. Explaining the figures to others also helped me realize exactly how much of the material I had understood.

Reflecting back on everything, I can't even imagine how everything was at the lab eight weeks ago. Time has definitely moved fast. Science, and research in particular, has tremendous potential in so many different areas of study. To think that in a few years, we may have gotten much closer to finding different alternatives and better success rates for organ transplants, or cured autoimmune diseases, is an amazing thing. Although research will inevitably lead to a lot of failure, it also leads to great success, and with more breakthroughs that will follow in time to come, we are sure to progress in treatment and care of humans. Everything here starts in the mouse, but success at the smaller animal level is what leads to clinical trials and success in people. I will really miss coming here and learning something new every single day. I've realized that what I love most is talking about the science behind the lab work, and how that thought process can generate new hypotheses that we can look forward to testing and growing on. It was a wonderful experience that I'm very grateful for, and I can only hope to come back to later on. Thank you!

Week Seven


I started off the week with two final ELISAs, and the data definitely confirmed things from the Elispot and FACS. It's great to finally have this data ready, because it proves the validity of this experiment, even though the data itself is unexpected and contradicts previous findings. Because of this, we will have to closely re-examine the previous experiments dealing with the complement receptors.
Journal Club, which we've been attending regularly, was on an especially interesting topic. We studied an article which explores the mechanisms why females are more prone to autoimmune diseases than males. The journal's findings suggest that the XX chromosome complement is what actually confers greater susceptibility to autoimmune diseases, such as lupus, as compared to the XY chromosome complement. The experiments themselves used castration techniques to focus on the susceptibility of chromosome complements alone, without the added variability factor of sex hormones. The study was mostly in vivo, and done in mice models. A hypothetical conclusion that could be derived has to do with their data showing that there is a higher expression of IL-13Ra2 on XX than on XY. IL-13Ra2 is only expressed on the X chromosome, and it can downregulate Th2 responses (which are CD4+ helper T cells regulating antibody-mediated immunity, a major part of the immune system). Figuring out the mechanics of the puzzle may eventually lead to a cure for severely debilitating autoimmune diseases - that would be wonderful.
By the end of the week, I performed a mini-takedown myself. I harvested and mashed the spleen, counted the cells, diluted and resuspended, and titrated the antibodies to find the correct titer for use in future experiments.
Overall, it was a great week. Summer is going by so quickly - only one more week here!