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目的比较C57BL/6小鼠肝脏、肺脏、脾脏和肠系膜淋巴结中γδT细胞占所分离的淋巴细胞及其CD3+T细胞的百分比、表型和功能的特点。方法分离正常C57BL/6小鼠肝脏、肺脏、脾脏和肠系膜淋巴结的淋巴细胞,应用细胞表面分子染色的方法,使用流式细胞仪观察γδT细胞占从不同组织分离的淋巴细胞及CD3+T细胞的百分比及其表型的特点。细胞经PMA和离子霉素刺激后,应用细胞内细胞因子染色的方法,通过流式细胞仪观察γδT细胞产生IFN-γ、IL-4、IL-9和IL-17细胞因子的情况。结果γδT细胞占分离淋巴细胞中的百分含量在肝脏明显高于肺脏、脾脏和肠系膜淋巴结(P<0.05),而γδT细胞在肠系膜淋巴结CD3+T细胞的百分含量要显著的低于其他脏器(P<0.05)。不同组织器官中γδT细胞以CD4-CD8-表型为主,还存在少量CD8+γδT细胞。在肠系膜淋巴结γδT细胞中CD4+细胞的量明显高于其他器官,且明显可见一群CD4+CD8+细胞。不同组织中γδT细胞中IL-17+的细胞量要明显高于IFN-γ+和IL-4+细胞。γδT细胞基本不分泌IL-9。肺脏的γδT细胞分泌细胞因子的能力最强,其IL-17+细胞的量达到(26.6±12.1)%。IFN-γ+细胞的量在肝脏和肺脏中较高,分别为(1.36±0.37)%和(1.6±0.7)%。结论 C57BL/6小鼠肝脏、肺脏、脾脏和肠系膜淋巴结γδT细胞的含量、表型和功能方面存在显著性差异。
Objective To compare the percentage, phenotype and function of γδT cells in lymphocytes and CD3 + T cells isolated from liver, lung, spleen and mesenteric lymph nodes of C57BL / 6 mice. Methods Lymphocytes in the liver, lung, spleen and mesenteric lymph nodes of normal C57BL / 6 mice were isolated and stained with cell surface molecule. Flow cytometry was used to observe the percentage of γδT cells in lymphocytes and CD3 + T cells isolated from different tissues Percentage and its phenotypic characteristics. The cells were stimulated with PMA and ionomycin, and the cytokine production of IFN-γ, IL-4, IL-9 and IL-17 by γδT cells was observed by flow cytometry after intracellular cytokine staining. Results The percentage of γδT cells in isolated lymphocytes in the liver was significantly higher than that in the lung, spleen and mesenteric lymph nodes (P <0.05), but the percentage of γδT cells in mesenteric lymph nodes was significantly lower than that in other organs (P <0.05). ΓδT cells in different tissues and organs mainly CD4 - CD8 phenotype, there are still a small amount of CD8 γ γ T cells. The amount of CD4 + cells in mesenteric lymph node γδT cells was significantly higher than that in other organs, and a group of CD4 + CD8 + cells were clearly seen. The amount of IL-17 + in γδT cells in different tissues was significantly higher than that in IFN-γ + and IL-4 + cells. γδ T cells do not substantially secrete IL-9. ΓδT cells in lungs had the highest ability to secrete cytokines, and the amount of IL-17 + cells reached (26.6 ± 12.1)%. The amount of IFN-γ + cells was higher in the liver and lung, respectively (1.36 ± 0.37)% and (1.6 ± 0.7)%. Conclusion The content, phenotype and function of γδT cells in liver, lung, spleen and mesenteric lymph nodes of C57BL / 6 mice are significantly different.