ROR-γt干扰腺病毒载体抑制Th17细胞功能

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

摘要:

目的：设计和构建ROR-γt特异性干扰腺病毒载体，观察其对大鼠脾脏来源的辅助性T 细胞17（T helper cell 17，Th17）功能的影响。方法：设计ROR-γt编码区干扰序列，DNA聚合酶作用下形成双链序列，与线性化pSES-HUS质粒连接。转入BJ5183，转染病毒包装体系293细胞系，收集细胞，得到病毒液。反复感染4次得到高滴度pSES-HUS-ROR-γt病毒液。将其感染大鼠脾脏来源的Th17细胞，检测其对Th17细胞功能的影响。结果：成功构建和包装pSES-HUS-ROR-γt干扰腺病毒液，Th17细胞感染pSES-HUS-ROR-γt干扰病毒后，其分泌细胞因子白介素（interleukin，IL）-17、IL-6和TNF-γ水平下降。结论：pSES-HUS-ROR-γt干扰腺病毒能够有效抑制Th17细胞功能，可能对肾移植慢性排斥反应具有抑制或者延缓作用。

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.