Mouse lymphocyte study

Dr. Paulson's group investigates the roles of carbohydrate-binding proteins involved in immune regulation and human disease, through their interaction with carbohydrate groups expressed on cell surface glycoconjugates. This laboratory is currently focused on how sialoside ligands modulate Siglec functions and on understanding the molecular basis of glycosylation changes following differentiation and activation of leukocytes. Evaluation of Glycosyltransferases and CBP expression in B cells, CD4+ and CD8+ T cells following activation. The overall goal of this study is to understand the molecular regulation of cell surface glycosylation in mouse lymphocytes before and after activation. Initial studies using lectins and antibodies showed differences in glycan expression following activation, which included a marked increase in PNA binding. Interestingly, this increase was observed in B, CD4+ and CD8+ cells after 72h of activation. Our aim is to elucidate the molecular basis for such changes in B and T lymphocytes (CD4+ and CD8+), focusing on the mRNA expression of glycogenes before and after activation of mixed spleenocytes. In brief, the experimental protocol is as follows: splenocytes are isolated from C57Bl/6 mice (6-10 wks old) and cultured for 0, 24, 48 and 72 hrs. Cultures containe either anti-CD3/IL2/IL4 or anti-IgM/IL4 to activate T cells and B cells respectively. At each time point, B, CD4+ and CD8+ cells are purified by positive selection using magnetic beads and total RNA is prepared from each cell type.