淋巴细胞转输对线粒体腺苷酸转位酶诱导的自身免疫性心肌病小鼠细胞因子的抑制作用

Transferred splenic lymphocytes suppressed cytokine expression in mice with experimental autoimmune cardiomyopathy induced by ANT peptides

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。

Objective：To explore the T helper-1 （Th1）/T helpe-2 （Th2） subset balance of splenic T cells, serum cy tokine levels, and myocardial cytokine mRNA levels in mice with experimental autoimmune cardiomyopathy induced by adenine nucleotide translocator （ANT） peptides after transferring toleranced lymphocytes. Method： Mice immunized several times with the synthesized ANT peptides were used as the cardiomyopathy （DCM） group. Mice in the anti-L3T4 McAb treated group were immunized with same peptides, followed by tail vein injection of 400 μg of anti-L3T4 on days 0, 1 and 2 post-immunization. On the 180th day, the splenic lymphocytes from the anti-L3T4 group were transferred into syngeneic recipients （transfer-group）. These mice were also immunized with same peptides as the DCM group. Control mice were immunized with the same solution as that injected into the DCM group, without the peptides. We examined the percentages of interferon （IFN-γ） and interleukin-4 （IL-4） producing cells in splenic CD4-positive lymphocytes using flow cytometry analysis. Serum IFN-γ, IL-2, IL-4, IL-6 and TNF-α levels were measured by enzyme-linked immunosorbent assay （ELISA） and the levels of myocardial cytokine were detected by reai-time PCR. Result： IFN-γ and IL-4 levels in Th cells in the transfer-group were almost the same as in the control and anti-L3T4 groups, which were significantly lower than the DCM group. In the transfer-group, serum IFN-γ and IL-2 levels were higher than the DCM group and lower than the anti-L3T4 group, while IL-4 and IL-6 levels were lower than the DCM group. TNF-α level significantly increased in transfer group as compared with the former three groups. Cytokine production in the myocardium was dramatically inhibited in the transfer and the anti-L3T4 groups, both of which were similar to the control group. Conclusion： These results suggest that cytokines production in mice immunized with ANT peptides may be suppressed by transferred splenic lymphocytes. Transferred splenic lymphocytes and anti-L3T4-antibody immunization produce a similar result.