结核病小鼠T淋巴细胞亚群及其表达的四项细胞因子分析

目的 探讨T淋巴细胞(简称T细胞)表达穿孔素(PFN)、颗粒酶B(GzmB)、γ-干扰素(IFN-γ)、白细胞介素-2(IL-2)与结核免疫的关系.方法 60只MK小鼠按随机数字表法分成结核病组和健康对照组,每组30只.以CD3PerCP、CD4FTTC、CD8APC单抗标记T细胞亚群,以藻红蛋白标记PFN、GzmB、IFN-γ、IL-2单抗标记细胞因子,以流式细胞仪检测分析总的淋巴细胞、CD3+、CD4+、CD8+、CD4+CD8+;双阳(DP)T细胞计数和各亚群占淋巴细胞百分率,观察各T细胞亚群内表达PFN、GzmB、IFN-γ、IL-2的阳性细胞计数和各自占淋巴细胞百分率.采用t检验进行统计学分析.结果 (1)CD4+CD8+、DP T细胞均能不同程度地表达PFN、GzmB、IFN-γ、IL-2.PFN、GzmB的表达以CD8+T细胞占优势.IFN-γ、IL-2的表达以CD4+T细胞占优势.(2)T细胞亚群细胞计数结果与T细胞亚群占总淋巴细胞百分率结果,所反映的T细胞免疫变化趋势可能相似,也可能不同甚至相反.(3)两组间表达PFN的T细胞差别不明显,但结核病组表达GzmB的各个T细胞亚群计数和CD3+%、CD8+%、DP%高于对照组(t值为-3.72～4.13.均P<0.05).(4)结核病组表达IFN-γ的CD3+、CD4+T细胞计数高于对照组;但结核病组表达IL-2的CD8+、DP T细胞计数和CD3+%、CD4+%、CD8+%、DP%均低于对照组(t值为2.62～3.46,均P<0.05).结论 、CD,4+、CD8+、DPT细胞均能不同程度地表达PFN、GzmB、IFN-γ、IL-2;联合评价T细胞各亚群计数和其占淋巴细胞百分率更能判断免疫学状态.

Analysis of T lymphocyte subsets and their expression of 4 cytokines in tuberculosis mice

Objective To investigate the relationship of anti-tuberculosis immunity with perforin (PFN), granzyme B( GzmB), interferon-γ (IFN-γ), and interleukin-2 (IL-2) expression by T lymphocyte subsets. Methods Sixty mice were randomly allocated into a tuberculosis group and a control group(n=30each). Surface markers of T lymphocytes were stained with CD3PerCP, CD4FTTC, CDsAPC, and intracellular cytotoxic molecules with PE-PFN, PE-GzmB, PE-IFN-γ, and PE-IL-2 multi-color-labeled monoclonal antibodies, and analyzed at the single cell level. The relation between T lymphocyte subsets expression PFN, GzmB, IFN-γ, IL-2 and antituberculosis immunity by flow cytometer. Results (1) CD4+,CD8+ and CD4+CD8+DP) T lymphocytes all expressed PFN, GzmB, IFN-γ and IL-2 to some degrees. The expressions of PFN and GzmB were much higher in CD8+T lymphocytes than those in CD4+T lymphocytes, while the expressions of IFN-γ and IL-2 were higher in CD4+T lymphocytes. (2) The counts of T lymphocyte subsets and the percentages of T lymphocyte subsets to total lymphocytes may or may not reflect the cellular immunity consistently. (3)There was no significant difference in T lymphocytes expressing PFN between the tuberculosis group and the control group. But the counts of CD3+.CD4+, DP and CD8+T lymphocytes and the percentages of CD3+ DP and CD8+cells expressing GzmB were significantly increased in the tuberculosis group( t value from -3.72 to 4. 13, all P<0.05 ). (4) IFN-γ expressing CD3+and CD4 lymphocytes were increased significantly in the tuberculosis group. The counts of CD8+and DP T lymphocytes and the percentages of CD3+, CD4+, CD8+, and DP cells that expressed IL-2 were decreased significantly in the tuberculosis group ( t value from 2.62 to 3.46, all P<0.05). Conclusion CD4+, CD8+and DP lymphocytes all can express PFN, GzmB, IFN-γand IL-2 at different degree levels.