Genes Differentially Expressed in Activated B Cell Isolated By Substractive cDNA Hybridisation
Posted on: Thursday, 17 June 2004, 06:00 CDT
CD40 is a cell surface receptor that belongs to the tumor necrosis factor-R (TNF-R) family, and that was first identified and functionally characterized on B lymphocytes. In recent years it has become clear that CD40 is expressed much more broadly, including expression on monocytes, dendritic cells, endothelial cells and epithelial cells. It is a 45-50 kDa glycoprotein of 277aa. CD40 binds to a ligand (CD40L also called CD154 or gp39) which is a 35 KDa glycoprotein of 261 aa, a member of the tumor necrosis factor family too. This CD40L is expressed on activated T cells, mostly CD4^sup +^ but also some CD8^sup +^ as well as basophils/mast cells. Extensive studies on CD40 activation of B cell in vivo have demonstrated that CD40 activation has major effects on many steps of the B cell natural history. CD40 activates proliferation, differentiation and Ig production of immature and mature B cell subsets. In addition, CD40 can induce re-expression of telomerase activity in memory B cells, thereby contributing to an expanded lifespan of these cells. CD40 activation guides the B cell through their differentiation program, including rescue from apoptosis, differentiation into germinal center cells, isotype switching, selection and maturation into memory cells. However, CD40 prevents the terminal differentiation of activated mature B cells into plasma cells. Most of these processes take place in specialized anatomical strucutres: germinal centers. To study the molecular basis of CD40 function, we used a cDNA subtraction approach. We report here on genes, Fig1 (interleukin-four induced gene 1), first characterized immediate-early IL4 inducible gene from B cells., CLAST2 (CD40 ligand activated specific transcripts) and BID (BH3 interacting domain death agonist). The effective differential expression of these genes was confirmed by northern blotting. We analysed expression of the gene under-investigation in germinal center cells from mice immunized with Ti and Tdepenclent antigens. To investigate the expression of these genes in different B cell development stages, we are perform an RT-PCR on different cell lines considered at different stage of differentiation. Finally we are producing monoclonal mouse antibodies against FIGl and CLAST2 proteins to be employed in differential protein expression studies.
E. Betto1,2, S. Merluzzi1,2, O. D'Orlando1,2, R. Bruno1,2 , S. Altamura1,2, C. Pucillo1,2
1 Department of Biomedical Sciences and Technologies, University of Ucline, Udine, Italy; 2 M.A.T.I Center of Excellence University of Udine, Udine, Italy
Copyright Edizioni Minerva Medica Mar 2004
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