【摘要】 目的以CD80、CD86抗体作为激动剂、脾细胞为研究对象,观察CD80、CD86抗体对耗竭性T细胞效应功能的影响。方法将42只8周龄雌性HLA-A11DR1转基因小鼠随机分为7组各6只,分别在8、11、14周龄时,A~F组臀部皮下接种磷脂酰肌醇蛋白聚糖3(GPC3)多肽,G组接种等体积无热源PBS;A~E组在24周龄和F、G组在17周龄时脱颈处死,取脾脏并制成脾细胞悬液;A、B组分别滴加20、40μg/m L抗鼠CD80抗体和10μg/m L GPC3多肽,C、D组分别滴加20、40μg/m L抗鼠CD86抗体和10μg/m L GPC3多肽,E、F、G组仅滴加10μg/m L GPC3多肽。孵育18 h后,采用酶联斑点分析仪检测干扰素γ(IFN-γ),以此判断T细胞效应功能。结果 A~G组脾细胞IFN-γ阳性斑点数分别为(80.61±48.91)、(207.67±60.41)、(1.67±0.97)、(1.33±0.49)、(2.33±1.53)、(38.17±5.18)、(2.33±1.53)个。其中,F组高于E、G组(P均<0.01),E、G组间比较无统计学差异;A组>B组>C、D、E组(P均<0.01),C、D、E组间比较差异无统计学意义;而且,IFN-γ阳性斑点数随着CD80抗体浓度升高而增加,二者呈正相关(r=0.760 5,P<0.01)。结论 CD80抗体能够刺激耗竭性T细胞恢复产生细胞因子,且呈剂量依赖的动力效应;而CD86抗体未能检测到该功能。更多还原

【Abstract】 Objective To investigate the effects of CD80 and CD86 antibody on the effector function of exhausted T cells. Methods Forty-two female HLA-A11DR1 transgenic mice were randomly divided into 7 groups(n = 6). The mice in groups A-F were subcutaneously inoculated with phosphatidylinositol 3(GPC3) at the age of 8 weeks(week 8),at week11 and week 14,respectively. Mice in the group G were inoculated with the same volume of heat-free PBS as controls.Mice in the groups A-E were sacrificed at week 24,and groups F and G at week 17. Splenocytes suspension was made.Then,20 or 40 μg/m L CD80 antibody and 10 μg/m L GPC3 peptide were pipetted in the splenocytes of groups A and B;20 or 40 μg/m L CD86 antibody and 10 μg/m L GPC3 were pipetted in the groups C and D,respectively; 10 μg/m L GPC3 peptide was pipetted in the groups E,F,and G. After incubation for 18 h,interferon-γ(IFN-γ) was measured by enzymelinked spot analyzer for analysis of T cell function. Results IFN-γ positive spots in the groups A-G were 80. 61 ± 48. 91,207. 67 ± 60. 41,1. 67 ± 0. 97,1. 33 ± 0. 49,2. 33 ± 1. 53,38. 17 ± 5. 18 and 2. 33 ± 1. 53,respectively. Among them,group F was significantly higher than groups E and G(both P < 0. 01). There was no significant difference between groups E and G(both P < 0. 01). Group A was more than group B,and group B was significantly more than group C,D and E(all P < 0. 01). There was no significant difference among groups C,D and E. In addition,the number of IFN-γ positive spots was positively correlated with CD80 antibody concentration(r = 0. 7605,P < 0. 01). Conclusions Anti-CD80 could restore the exhausted T cells to secret IFN-γ with a dose-dependent manner,while anti-CD86 could not be detected for the same effect.更多还原