Construction of ROR-γt interference adenovirus and its influence on Th17 cell function

Abstract:

Objective：To construct the ROR-γt interference adenovirus，to transfect the ROR-γt interference adenovirus into T helper cell 17（Th17 cell） and to detect the change of Th17 cell was after being affected by ROR-γt interference adenovirus. Methods： ROR-γt interference sequences were designed to form a double chain sequence by DNA polymerase and to connect with the linearized pSES-HUS plasmid. pSES-HUS-ROR-γt plasmid was transfected into BJ5183 by electroporation，and BJ5183 was transfected into the 293 virus package system. The ROR-γt interference adenovirus vector was achieved and named as pSES-HUS-ROR-γt. pSES-HUS-ROR-γt was infected the Th17 cell which was isolated from the spleen of rat. The changes of IL-17，IL-6 and TNF-γ were detected by ELISA. Results：pSES-HUS-ROR-γt interference adenovirus vector was successfully constructed. Th17 cytokines（IL-17，IL-6 and TNF-γ） were decreased after Th17 being infected with pSES-HUS-ROR-γt interference adenovirus. Conclusion： Th17 cell function can be inhibited by pSES-HUS-ROR-γt interference adenovirus vector，and the adenovirus vector has the potential treatment function in the chronic rejection of renal transplantion.