【摘要】 目的 探讨肿瘤坏死因子α诱导蛋白8样分子2（TIPE2）在T细胞发育过程中的表达及作用，分析其在哮喘发病中的潜在作用。方法 选择C57BL/6J雌性野生型（WT）小鼠、C57BL/6J雄性TIPE2基因敲除(Tipe2^-/-)小鼠、C57BL/6J雄性绿色荧光蛋白敲入(Tipe2^gfp/+)小鼠为研究对象。取WT和Tipe2^gfp/+小鼠结肠组织和胸腺组织，获取肠道固有层淋巴细胞（LPLs）和胸腺细胞，采用流式细胞术检测结肠组织CD45^-细胞、CD45⁺细胞及双阴性（DN）T细胞、双阳性（DP）T细胞、CD4⁺T细胞和CD8⁺ T细胞中TIPE2的表达。取WT和Tipe2^-/-小鼠外周血、脾脏和肠系膜淋巴结，采用流式细胞术检测外周血、脾脏和肠系膜淋巴结中CD4⁺和CD8⁺T细胞水平。将10只Tipe2^gfp/+小鼠随机分为对照组和哮喘组，每组5只。哮喘组小鼠以卵清蛋白诱导哮喘模型，对照组小鼠以生理盐水代替卵清蛋白诱导；采用流式细胞术检测2组小鼠肺泡灌洗液中CD4⁺T细胞和CD8⁺T细胞中TIPE2的表达。结果 WT和Tipe2^gfp/+小鼠结肠组织LPLs中，大部分TIPE2⁺细胞表达免疫细胞的标志分子CD45,几乎所有的CD45^-基质细胞不表达TIPE2。Tipe2^gfp/+小鼠结肠组织中基质细胞和WT小鼠免疫细胞中检测不到TIPE2⁺细胞。Tipe2^gfp/+小鼠胸腺组织DN T细胞中TIPE2相对表达量显著低于CD4⁺T细胞和CD8⁺ T细胞（t=53.312、8.230,P<0.01）;DP T细胞中TIPE2相对表达量显著低于CD4⁺T细胞和CD8⁺ T细胞（t=21.094、6.405,P<0.001）;DN T细胞中TIPE2相对表达量显著低于DP T细胞（t=7.619,P<0.01）;CD4⁺T细胞与CD8⁺ T细胞中TIPE2相对表达量比较差异无统计学意义（t=1.256,P>0.05）。WT小鼠与Tipe2^-/-小鼠外周血、脾脏和淋巴结中CD4⁺T细胞百分比比较差异无统计学意义（t=0.670、0.128、0.128,P>0.05）;WT小鼠与Tipe2^-/-小鼠外周血及脾脏和淋巴结中CD8⁺ T细胞百分比比较差异无统计学意义（t=1.488、0.542、0.255,P>0.05）。哮喘组小鼠肺泡灌洗液CD4⁺T细胞中TIPE2相对表达量显著低于对照组（t=15.370,P<0.001）;对照组与哮喘组小鼠肺泡灌洗液CD8⁺ T细胞中TIPE2相对表达量比较差异无统计学意义（t=0.132,P>0.05）。结论 在T细胞的不同发育阶段TIPE2的表达水平不同；TIPE2基因缺陷对T细胞的发育过程无明显影响，可显著影响哮喘发生发展过程中CD4⁺T细胞功能。更多还原

【Abstract】 Objective To investigate the the expression and role of tumor necrosis factor-α inducible protein 8 like molecule 2（TIPE2） in the development of T cells, and analyze its potential role in the pathogenesis of asthma.Methods C57 BL/6 J female wild-type（WT） mice, C57 BL/6 J male Tape2^-/- mice, and C57 BL/6 J male Tape2^gfp+ mice were selected as research objects.The colon tissue and thymus tissue of WT and Tipe2^gfp/+mice were taken to obtain intestinal lamina propria lymphocytes（LPLs） and thymocytes.The expression of TIPE2 in CD45 cells, CD45⁺cells, double negative（DN） T cells, double positive（DP） T cells, CD4⁺T cells and CD8⁺T cells was detected by flow cytometry.The levels of CD4⁺ T cells and CD8⁺T cells in peripheral blood, spleen and mesenteric lymph nodes of WT and Tipe2^-/- mice were detected by flow cytometry.A total of 10 Tipe2^gfp/+ mice were divided into control group and asthma group, with 5 mice in each group.Asthma model of mice was induced by ovalbumin in the asthma group, and normal saline was replace ovalbumin in mice in the control group; the expression of TIPE2 in CD4⁺ T cells and CD8⁺ T cells in alveolar lavage fluid of mice in the two groups was detected by flow cytometry.Results In the LPLs in colon tissues of WT and Tipe2^gfp/+ mice, most TIPE2⁺ cells expressed the marker of immune cells CD45^-,and almost all CD45^- stromal cells did not express TIPE2.TIPE2⁺ cells were not detected in stroma cells in colon tissue of Tipe2^gfp/+ mice and immune cells of WT mice.The relative expression level of TIPE2 in DN T cells was significantly lower than that in CD4⁺ T cells and CD8⁺ T cells in thymus of Tipe2^gfp/+ mice（t=53.312,8.230;P<0.01）;the relative expression level of TIPE2 in DP T cells was significantly lower than that in CD4⁺ T cells and CD8⁺ T cells（t=21.094,6.405;P<0.001）;the relative expression level of TIPE2 in DN T cells was significantly lower than that in DP T cells（t=7.619,P<0.01）;there was no significant difference in the relative expression level of TIPE2 between CD4⁺ T cells and CD8⁺ T cells（t=1.256,P>0.05）.There was no significant difference in the percentage of CD4⁺ T cells in the peripheral blood, spleen and lymph nodes between WT mice and Tipe2^-/- mice（t=0.670,0.128,0.128;P>0.05）;there was no significant difference in the percentage of CD8⁺ T cells in the peripheral blood, spleen and lymph nodes between WT mice and Tipe2^-/- mice（t=1.488,0.542,0.255;P>0.05）.The relative expression level of TIPE2 in CD4⁺ T cells in alveolar lavage fluid of mice in the asthma group was significantly lower than that in the control group（t=15.370,P<0.001）;there was no significant difference in the relative expression level of TIPE2 in CD8⁺ T cells of alveolar lavage fluid between the control group and asthma group（t=0.132,P>0.05）.Conclusion The expression level of TIPE2 are different in the different developmental stages of T cells.TIPE2 gene deficiency does not significantly affect the development of T cells, it can significantly affect the CD4⁺ T cells function in the pathogenesis of asthma.更多还原