ICOS联合IFN-γ单抗对活化T细胞诱导的RINm5F细胞凋亡的保护作用  

Protection against apoptosis of anti-ICOS combined with anti-IFN-γ monoclonal antibodies on activated T cell-induced RINm5F cells

作　　者：徐宽枫[1] 董丽[1] 刘翠萍[1] 武晓泓[1] 茅晓东[1] 刘超[1] 

机构地区：[1]南京医科大学第一附属医院内分泌科,210029

出　　处：《中华内分泌代谢杂志》2008年第1期39-42,共4页Chinese Journal of Endocrinology and Metabolism

基　　金：江苏省政府医学重点学科资助项目（SK200214）

摘　　要：目的 探讨可诱导性协同刺激分子（ICOS）单抗和γ-干扰素（IFN-γ）单抗联合应用对T细胞活化诱导的胰岛细胞凋亡及其相关凋亡机制。方法植物血凝素（PHA）活化的T细胞与胰岛细胞株RINm5F共同培养，联合应用ICOS单抗和IFN-γ单抗48h后，AnnexinV／PI法观察RINm5F细胞凋亡率，RT—PCR法观察凋亡相关和胰岛功能相关基因表达，放射免疫法检测观察胰岛素分泌水平。结果ICOS单抗与IFN-γ单抗联用，T细胞活化诱导的RINmSF细胞株凋亡率（％）显著下降（IgG对照71．89±3．29，ICOS单抗39．66±2．05，IFN-γ单抗45．65±1．59，ICOS单抗＋IFN-γ单抗20．49±0．95），这可能与死亡受体途径FAS／FAS配体，肿瘤坏死因子及其受体和肿瘤坏死因子相关凋亡诱导配体等相关的基因表达下调有关；同时两者联合应用后，RINm5F细胞株INS1、INS2及PDX-1基因表达量显著上调，胰岛素分泌水平也显著升高。结论ICOS单抗和IFN-γ单抗联合应用降低胰岛细胞凋亡的同时，对胰岛细胞的胰岛素分泌功能具有一定的保护作用，是一种有效的诱导T细胞对胰岛细胞无反应性的手段。Objective To investigate the effect of combining anti-ICOS （ inducible costimulator） and antiinterferon-γ （IFN-γ） on activated T cell-induced islet cell apoptosis and related gene expressions of the apoptosis pathway. Methods After adding anti-ICOS and anti-IFN-γ, PHA-activated T cells and RINm5F cells （ islet cell line） were cultured together for 48 h, the apoptosis rate of RINm5F cells was assayed by Annexin V/PI staining, and expressions of death receptor pathway related genes, INS1, INS2 and PDX-1 genes were determined by RTPCR The level of insulin secreted from RINm5F cells was measured by radioimmunoassay. Results Combined use of anti-ICOS and anti-IFN-γ significantly reduced apoptosis rate （ % ） of RINm5F induced by activated T cell （ igG control ： 71.89 ± 3.29, anti-ICOS： 39.66 ± 2.05, anti-IFN-γ： 45.65± 1.59, anti-ICOS ＋ anti-IFN-γ： 20. 49 ± 0.95）, which was related to down-regulation of FAS/FASL, tumor necrosis factor and its receptor and tumor necrosis factor-related apoptosis-inducing ligand pathway related genes. Furthermore the expressions of INS1, INS2 and PDX-1 genes and the secretion of insulin were increased in these cells. Conclusion The combined use of anti- ICOS and anti-IFN-γ not only protects RINm5F cells against apoptosis but also enhances insulin secretion of these cells.

关 键 词：RINm5F胰岛细胞 T细胞 可诱导性协同刺激分子 细胞凋亡 

分 类 号：R587.1[医药卫生—内分泌] R392.12[医药卫生—内科学]