IL-36γ促进人CD8^+T淋巴细胞细胞因子的分泌  

IL-36γ enhances human CD8^+T lymphocytes activation and promotes cytokines production

作　　者：吕全省 马静姝 古彦铮[2] 王雪峰[1] 王明元[3] 张学光[1] 朱一蓓[1] 卢斌峰[1] 

机构地区：[1]苏州大学基础医学与生命科学学院,江苏苏州215123 [2]苏州大学附属第一医院临床免疫研究所,江苏苏州215006 [3]苏州市中心血站输血研究室,江苏苏州215006

出　　处：《江苏大学学报（医学版）》2016年第3期193-198,共6页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(81273208;31428005);苏州市科技发展计划项目(SYS201424)

摘　　要：目的:探讨白细胞介素36γ(IL-36γ)是否能够增强体外培养的人CD8+T淋巴细胞分泌γ干扰素。方法:经密度梯度离心法从健康人外周血分离获得外周血单个核细胞(PBMC),经磁珠阳性选择富集纯化CD8^+T淋巴细胞,分别用CD3单克隆抗体(m Ab)+CD28 m Ab,CD3 mAb+CD28 mAb+IL-12,CD3 mAb+CD28 mAb+IL-36γ以及CD3m Ab+CD28 m Ab+IL-12+IL-36γ4种因子组合刺激培养24 h和72 h。收集细胞,采用免疫荧光标记和流式细胞术分析分选富集的CD8^+T淋巴细胞纯度以及γ干扰素的表达;采用实时PCR检测各组细胞IL-36受体(IL-36R)、γ干扰素和颗粒酶B mRNA的表达。结果:人CD8+T淋巴细胞表达IL-36R;各组因子刺激的人CD8^+T淋巴细胞形态没有明显差异;人CD8^+T淋巴细胞经IL-36γ刺激后,γ干扰素、颗粒酶B mRNA表达上调;同时,IL-36γ和IL-12具有协同刺激CD8+T淋巴细胞的作用。结论:IL-36γ能促进体外培养的人CD8^+T淋巴细胞分泌细胞因子,并且与IL-12具有协同作用。Objective： To investigate whether recombinant interleukin-36γ（rIL-36γ） can enhance hu- man CD8 ＋ T lymphocytes secrete cytokine interferon-γ （IFN-γ） in vitro. Methods： Peripheral blood mono- nuclear cells（PBMC） was isolated from healthy donors by density gradient centrifugation, and CD8 ＋T lym- phocytes were purified by positive magnetic beads selection from PBMCs. CD8 ＋ T cells were stimulated with different combination of cytokines（CD3 mAb ＋ CD28 mAb;CD3 mAb ＋ CD28 mAb ＋ IL-12; CD3 mAb ＋ CD28 mAb ＋ IL-36γ;CD3 mAb ＋ CD28 mAb ＋ IL-12 ＋ IL-36γ） in vitro. The cells of each group were col- lected after 24 h and 72 h. CD8 ＋ T lymphocytes purity and cytokine IFN-γproduction were determined by flow cytometry. Real time PCR （qRT-PCR） was used to detect IL-36 receptor （IL-36R）, IFN-γ and granzyme B mRNA expression. Results： Human CD8 ＋T lymphocytes expressed IL-36 receptor; there was no significant difference of cell morphology among the four groups of CD8 ＋ T lymphocytes; IFN-γ and granzyme B of CD8 ＋T lymphocytes stimulated by IL-36 γ were significantly increased in mRNA levels（P 〈 0.05） ;at the same time, IL-36γ and IL-12 had a costimulation role to activate CD8 ＋T lymphocytes. Con- clusion： IL-36γ has a synergistic effect with IL-12 to enhance human CD8 ＋T lymphocytes cytokine secre- tion in vitro.

关 键 词：白细胞介素36γ CD8+T淋巴细胞 Γ干扰素 颗粒酶B 

分 类 号：R392[医药卫生—免疫学]