From: ZA Yazici, Pathology & Microbiology (Z.A.Yazici@bristol.ac.uk)
Date: Sat Nov 08 2003 - 16:09:08 EST
Dear Flowers, I have been analysing murine splenocytes for their intracellular IL-2 expression within the CD4 positive (TCR transgenic) population. As expected, there is proliferation of CD4+ cells plus terget epitope, and minimal or no proliferation with medium alone. The supernatants of the proliferating cultures contain high levels of IL-2 as measured by ELISA. As can be seen in the attached powerpoint slides, when I look at the events by FACS analysis after staining with flourochrome conjugated anti-CD4 (FITC) and anti-IL-2 (PE), the proliferating events shift towards the double positive region in a diagonal fashion. (The blasting events become CD4 "bright".) My interpretation of the data is that the blasting events become (weakly) IL-2 positive as they become CD4 bright. However, the diagonality of the cycling cells can also mean that the compensations are not set correctly and that what seems weakly positive is actually increased autofluorescence of blasting cells. (There is no isotype control or FcR blocking in the assays). What are your comments? Thanks in advance. Zihni ---------------------- ZA Yazici (PhD), Pathology & Microbiology Department, School of Medical Sciences, University of Bristol, University Walk, Bristol, BS8 1TD, UK. Phone (W): +441179267584
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