不同时期应用抗L3T4单抗治疗的自身免疫性心肌病小鼠免疫球蛋白G亚型分析

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目的:探讨不同阶段应用抗L3T4单抗治疗自身免疫性心肌病对小鼠血清抗线粒体ADP/ATP载体抗体和免疫球蛋白G(IgG)亚型的影响。方法:将24只小鼠随机分为4组,每组6只。用含有人线粒体ADP/ATP载体肽的免疫液免疫近交系Balb/C小鼠建立类扩张型心肌病(DCM)模型(DCM组);以不含肽免疫液免疫小鼠作为对照组;在同时用含有ADP/ATP载体肽的免疫液的第0、1、2天连续3d以400μg抗L3T4单抗免疫小鼠获得早期治疗组;在给予ADP/ATP载体肽的免疫液第4个月初始连续3d给予抗L3T4单抗治疗获得中期治疗组。以ELISA法检测其血清中抗ADP/ATP载体抗体水平和IgG亚型(IgG1、IgG2a、IgG2b和IgG3)的变化;采用流式细胞术检测小鼠脾脏中CD4+T细胞内γ干扰素(IFN-γ)/白细胞介素(IL)-4的表达。结果:早期治疗组与对照组小鼠抗ADP/ATP载体抗体检测均为阴性,而中期治疗组与DCM组抗体均为阳性。DCM组各亚类水平均较对照组明显升高;早期治疗组IgG1、IgG2b和IgG3的产生均受到抑制,其水平接近对照组;中期治疗组IgG1水平介于DCM组和早期治疗组之间,IgG2b和IgG3的生成与DCM组差异无统计学意义,但显著高于早期治疗组;而治疗组与DCM组IgG2a水平则差异无统计学意义。早期治疗组小鼠IFN-γ/IL-4百分含量与对照组相比差异无统计学意义,但明显低于DCM组;中期治疗组IFN-γ的百分含量较DCM组更高,IL-4的百分含量介于DCM组和早期治疗组之间。结论:抗L3T4单抗治疗能够抑制DCM小鼠血清中的抗ADP/ATP载体抗体及IgG1亚型的生成和Th2相关细胞因子IL-4的百分含量,且以早期治疗的效果更明显。 Objective: To investigate the effects of anti-L3T4 monoclonal antibody on anti-mitochondrial ADP / ATP carrier antibody and immunoglobulin G (IgG) subtype in mice treated with autoimmune cardiomyopathy at different stages. Methods: 24 mice were randomly divided into 4 groups with 6 mice in each group. The model of dilated cardiomyopathy (DCM) was established by immunizing inbred Balb / C mice with the immunogenic solution containing human mitochondrial ADP / ATP carrier peptide. The mice immunized with peptide-free immunized solution were used as the control group. At the same time, mice were immunized with 400μg of anti-L3T4 monoclonal antibody on days 0, 1 and 2 of the ADP / ATP carrier peptide-containing immunization solution for three consecutive days to get the early treatment group. At the beginning of the fourth month of immunization with ADP / ATP carrier peptide Three consecutive anti-L3T4 mAbs were given to the medium-term treatment group. The serum levels of anti-ADP / ATP carrier antibody and IgG subtypes (IgG1, IgG2a, IgG2b and IgG3) were detected by ELISA. The levels of IFN-γ and IFN-γ in CD4 + T cells were detected by flow cytometry -γ) / Interleukin (IL) -4 expression. Results: The detection of anti-ADP / ATP carrier antibody in the early treatment group and the control group was negative, while the antibody in the intermediate treatment group and the DCM group were both positive. The level of each subclass of DCM group was significantly higher than that of the control group. The production of IgG1, IgG2b and IgG3 in the early treatment group was inhibited, and the level was close to the control group. The IgG1 level in the intermediate treatment group was between the DCM group and the early treatment group, The formation of IgG2b and IgG3 was not significantly different from that of DCM group, but significantly higher than that of the early treatment group. There was no significant difference in IgG2a level between the treatment group and the DCM group. The percentage of IFN-γ / IL-4 in the early treatment group was not significantly different from that in the control group, but significantly lower than that in the DCM group. The percentage of IFN-γ in the intermediate treatment group was higher than that in the DCM group -4 was between the DCM group and the early treatment group. Conclusion: Anti-L3T4 monoclonal antibody treatment can inhibit the generation of anti-ADP / ATP carrier antibody and IgG1 subtype and the percentage of Th2-related cytokine IL-4 in serum of DCM mice, and the effect of early treatment is more obvious.
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