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目的:研究淋巴细胞转输对线粒体腺苷酸转位酶(ANT)合成肽诱导的自身免疫性心肌病传染性耐受的小鼠Th1/Th2亚群及血清和心肌组织细胞因子的影响。方法:用ANT多肽多次免疫Balb/C小鼠得到心肌病组,单抗组小鼠在给予ADP/ATP肽免疫的第0、1和2天,同时接受尾静脉注射400μg抗L3T4单抗;第6个月末时,将单抗组小鼠的脾细胞取出转输到正常小鼠体内(转输组),此小鼠亦同时给予多肽免疫,方法同心肌病组。对照组以不含ANT的相同免疫进行假性免疫,时间和剂量同心肌病组。采用流式细胞术检测脾T细胞内细胞因子IFN-γ/IL-4的含量;以ELISA法检测其血清中IFN-γ、白细胞介素-2(IL-2)、IL-4、IL-6和肿瘤坏死因子(TNF-α)水平;实时荧光定量PCR法检测其心肌细胞因子mRNA表达。结果:转输组Th细胞IFN-γ、IL-4含量与对照组和单抗组接近且明显低于心肌病组;转输组小鼠血清IFN-γ和IL-2水平明显高于心肌病组而低于单抗组,IL-4和IL-6水平显著低于心肌病组;TNF-α水平在转输组则最高;心肌细胞因子mRNA的表达则转输组显著低于心肌病组,与对照组和单抗组相近。结论:淋巴细胞转输能够阻断心肌病诱导过程中绝大部分细胞因子的生成,与单抗早期干预的结果类似。
AIM: To investigate the effects of lymphocyte transfusion on Th1 / Th2 subsets and cytokines in serum and myocardium of mice tolerant to mitochondrial adenylate translocase (ANT) synthetic peptide-induced autoimmune cardiomyopathy. Methods: Balb / C mice were immunized with ANT polypeptide for multiple times to get cardiomyopathy group. Mice were immunized with ADP / ATP peptide on days 0, 1 and 2, and 400 μg of anti - L3T4 mAb was injected via tail vein. At the end of the 6th month, the splenocytes of Mab-Mice were removed and transferred to normal mice (transfusion group). The mice were immunized with the same peptide at the same time. The control group was immunized with the same immunity without ANT for the same time and dose of the cardiomyopathy group. The levels of cytokine IFN-γ / IL-4 in splenic T cells were detected by flow cytometry. The levels of IFN-γ, IL-2, IL-4 and IL- 6 and tumor necrosis factor (TNF-α) were detected by real-time PCR. The mRNA expression of cardiac cytokines was detected by real-time fluorescence quantitative PCR. Results: The levels of IFN-γ and IL-4 in the Th cells of the transfused group were significantly lower than those in the control group and the monoclonal antibody group, while the levels of IFN-γ and IL-2 in the transfused group were significantly higher than those of the cardiomyopathy IL-4 and IL-6 levels were significantly lower than those in the cardiomyopathy group, TNF-αlevel was the highest in the transfusion group, and the expression of myocardial cytokines mRNA in the transfusion group was significantly lower than that in the cardiomyopathy group , Similar to the control and mAb groups. CONCLUSIONS: Lymphocyte translocation blocks the generation of most cytokines during the induction of cardiomyopathy, similar to the results of early intervention with monoclonal antibodies.