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Investigation of apoptotic related genes in SEB induced human PBMC module
Author(s) -
Mendis Chanaka,
Jett Marti
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.648.5
Subject(s) - gene , apoptosis , gene expression , peripheral blood mononuclear cell , biology , caspase , microbiology and biotechnology , programmed cell death , genetics , in vitro
Our study is to characterize a set of apoptotic related genes that are key in the pathogenesis of Staphylococcal Enterotoxin B (SEB) induced lethal shock in human peripheral blood mononuclear cells (PBMC) and then utilize the gene expression pattern to further investigate the effectiveness of potential signal blockers. By using a set of genes previously identified through differential display (DD) and micro‐array techniques we carried out a conformational study of SEB induced apoptotic events in the following manner. First, we carried‐out an extensive literature search to characterize apoptosis related genes identified by DD and micro‐arrays. Second, we designed primers using stringent parameters, third we confirmed the gene expression pattern using RT‐PCR and then validated the gene expression patterns by further investigation of protein expression using ELISA assays and finally utilized the known gene expression patterns to evaluate the efficacy of potential signal blockers. We were able to confirm the previously observed gene expression patterns of heparanase precursor, Caspase 3, Caspase 6, Caspase 7, Caspase 8, Ubiquitin specific protease and SOD genes through RT‐PCR, protein expression pattern of a set of caspases through ELISA assays and correlated the functions of genes to SEB induced apoptosis related events. Our study also confirmed the effectiveness of the inhibitors as potential target to block SEB induced apoptotic events. We believe that our investigation may reveal crucial information to better understand the mechanisms as well as the regulation/inhibition of SEB induced apoptotic events.