CD80、CD86抗体对耗竭性T细胞效应功能的影响

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、40 μg/mL 抗鼠CD80抗体和10 μg/mL GPC3多肽,C、D组分别滴加20、40 μg/mL 抗鼠CD86抗体和10 μg/mL GPC3多肽,E、F、G组仅滴加10 μg/mL 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.%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 week 11 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/mL CD80 antibody and 10 μg/mL GPC3 peptide were pipetted in the splenocytes of groups A and B;20 or 40 μg/mL CD86 antibody and 10 μg/mL GPC3 were pipetted in the groups C and D, respectively;10 μg/mL GPC3 peptide was pipetted in the groups E, F, and G.After incubation for 18 h, interferon-γ (IFN-γ) was measured by enzyme-linked 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.