模拟微重力对肺T淋巴细胞激活功能的影响

发布时间：2018-05-14 23:00

本文选题：模拟微重力 + 旋转生物反应器　；参考：《免疫学杂志》2007年05期

【摘要】：目的探讨模拟微重力对肺T淋巴细胞活化功能的影响及机理。方法分离肺T淋巴细胞,利用的旋转式细胞组织培养系统(RCCS,即旋转生物反应器)模拟微重力条件。培养T淋巴细胞后,采用PepTag非放射性PKC检测试剂盒测定PKC活性;采用免疫组织化学染色观察表达核因子-κB(NF-κB);酶联免疫吸附技术(双抗体夹心ELISA法)测定白介素-2(IL-2)生成;采用流式细胞技术检测表面抗原CD25。结果普通细胞培养容器组培养的T淋巴细胞,刀豆素蛋白A(ConA)组与对照组比较,T淋巴细胞PKC活性增强,NF-κB的免疫组化染色阳性细胞的百分比增加,IL-2生成增加,表面表达CD25的细胞增多(P0.01);模拟微重力条件下,与普通细胞培养容器培养的T淋巴细胞相比较,ConA刺激后,PKC活性降低,NF-κB免疫组化染色阳性细胞百分比减低,IL-2生成下降,表面表达CD25细胞减少(P0.01);模拟微重力条件下,加入PKC特异的激动剂PMA,和ConA共同培养的T淋巴细胞组,与同样条件下ConA组相比,PKC活性略增强、NF-κB的免疫组化染色阳性细胞百分比、IL-2生成的量、表面表达CD25增加(P0.01)。结论模拟微重力条件下,T淋巴细胞的激活受到抑制;PKC的活性降低,表达NF-κB细胞数减少,表面表达CD25的细胞数降低,T淋巴细胞分泌的IL-2降低;同样条件下,PMA对T淋巴细胞的上述指标有部分恢复作用。
[Abstract]:Objective to investigate the effect and mechanism of simulated microgravity on the activation of lung T lymphocytes. Methods Lung T lymphocytes were isolated and the rotating cell tissue culture system (RCCS) was used to simulate microgravity conditions. After T lymphocytes were cultured, the activity of PKC was detected by PepTag non-radioactive PKC assay kit, the expression of nuclear factor- 魏 B (NF- 魏 B) was observed by immunohistochemical staining, the production of interleukin-2 (IL-2) was detected by enzyme-linked immunosorbent assay (Elisa). Surface antigen CD25 was detected by flow cytometry. Results compared with the control group, the percentage of PKC activity of T lymphocytes in the normal cell culture container group and concanavalin A Cona group was increased, and the percentage of the positive cells of NF- 魏 B immunocytochemical staining increased with the increase of IL-2 production. The number of cells expressing CD25 on the surface was increased, and the percentage of CD25 positive cells decreased after stimulation with Cona compared with that of T lymphocytes cultured in normal cell culture container under simulated microgravity, and the production of IL-2 was decreased in the cells that were positive for NF- 魏 B immunohistochemical staining. The surface expression of CD25 cells was reduced by P0.01G, and PKC specific agonist PMA and ConA co-cultured T lymphocytes were added under simulated microgravity. Compared with the same condition, the activity of ConA increased slightly the percentage of positive cells of NF- 魏 B and the amount of IL-2, and the expression of CD25 on the surface increased P0.01. Conclusion under simulated microgravity, the activation of T lymphocytes was inhibited, the number of NF- 魏 B cells was decreased, the number of cells expressing CD25 was decreased, and the IL-2 secreted by T lymphocytes was decreased. Under the same conditions, PMA can partially restore the above indexes of T lymphocytes.
【作者单位】：武警总医院博士后流动站中国人民解放军总医院南楼呼吸科武警总医院医学急救中心武警总医院呼吸科武警总医院核医学科
【分类号】：R85

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