Role of DR5 in TRAIL induced apoptosis of Jurkat cells

    BAI Huiª²Ling, GONG Feiª²Li, MA Yuanª²Fang

    1Department of Immunology Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China, 2Laboratory of Cellular and Molecular Immunology, Medical College , Henan University, Kaifeng 475001, China

    ¡¾Abstract¡¿ AIM: To study the role of DR5 in TRAIL apoptotic signal transduction in Jurkat cells. METHODS: cDNA containing fullª²length extracellular domain of human DR5 was cloned into pGAPZ¦Á. Recombinant Pichia pastoris clone generated via homologous recombination secreted high levels of sDR5. BALB/c mice were immunized with sDR5 in CFA to prepare antiª²DR5 mAb. The binding of antiª²DR5 mAb was measured for surface expression of TRAIL receptor by flow cytometric analysis. Jurkat cells were tested for their susceptibility to apoptosis induced by TRAIL with TRAIL apoptosis kit so as to study the correlation between DR5 expression level and TRAIL sensitivity. The level of DR5 expression on Jurkat cell line was examined by flow cytometry. The rates of TRAILª²induced apoptosis and antiª²DR5 mAb blocking on Jurkat cells were tested by flow cytometry with TRAIL apoptosis kit. RESULTS: The killing role of TRAIL was blocked in Jurkat cells pretreated with antiª²DR5 mAb. The average percentage of blocking was 90.49%. CONCLUSION: Antiª²DR5 mAb can specifically bind to DR5 and DR5 is expressed at high levels on Jurkat cells. DR5 plays a very key role in TRAIL induced apoptosis in Jurkat cells. DR5 expression is important for the induction of apoptosis by TRAIL. ......