输注负载胎盘滋养层细胞抗原的受者aaDC延长小鼠移植心的存活时间

目的 探讨输注负载供者胎盘滋养层细胞抗原的受者耐受性选择性激活树突状细胞(aaDC)对小鼠移植心存活时间的影响.方法 雌性Balb/c小鼠和雄性C57BL/6小鼠合笼后,获取雌鼠胎盘滋养层细胞,并提取其抗原.体外制备Balb/c小鼠的aaDC,分别与滋养层细胞抗原或者C57BL/6小鼠脾细胞抗原共培养,获得负载滋养层细胞抗原或者C57BL/6小鼠脾细胞抗原的aaDC,测定培养液中白细胞介素10(IL-10)和IL-12的含量以及aaDC表面CD40、CD80、CD86和主要组织相容性复合物(MHC)Ⅱ类分子的表达.取Balb/c小鼠,分别接受负载滋养层细胞抗原或者C57BL/6小鼠脾细胞抗原的aaDC输注(滋养层细胞抗原组和脾细胞抗原组),7 d后接受C57BL/6小鼠心脏异位移植,以注射生理盐水者为对照组,输注负载滋养层细胞抗原aaDC后接受昆明小鼠心脏移植者为第三供者对照组.术后记录移植心存活时间,并检测受者脾脏和移植心组织中IL-2、IL-10和γ干扰素(IFN-7)mRNA的表达.结果 负载脾细胞抗原的aaDC,CD86和MHCⅡ类分子的表达明显升高,而负载滋养层细胞抗原的aaDC,仅MHCⅡ类分子表达升高.滋养层细胞抗原aalDC的培养液中,IL-10为(122.6±15.4)ng/L,IL-12为(232.8±25.4)ng/L,空白对照aaDC的IL-10和IL-12分别为(35.4±8.5)ng/L和(267.3±12.5)ng/L,二者间IL-10的差异有统计学意义(P<0.05),而IL-12的差异无统计学意义(P>0.05).脾细胞抗原aaDC培养液中的IL-10和IL-12与空白对照aaDC相近,差异均无统计学意义(P>0.05).对照组移植心存活时间为(6.73±0.58)d,脾细胞抗原组为(12.28±0.76)d,滋养层细胞抗原组为(38.83±2.79)d,第三供者对照组为(6.68±0.62)d.滋养层细胞抗原组移植心存活时间明显长于脾细胞抗原组和对照组(P<0.01),而第三供者对照组和对照组间移植心存活时间的差异无统计学意义(P>0.05).结论 预先输注负载供者胎盘滋养层细胞抗原的受者aaDC,可延长小鼠移植心的存活时间.

Infusion of recipient alternative activated dendritic cells loaded with trophoblastic cells antigen prolonged survival of heart allografts

Objective To investigate the effect of recipient alternative activated dendritic cells (aaDC) loaded with trophoblastic cells antigen (TbC-Ag) on murine cardiac allograft survival Methods Female Balb/c mice were caged and mated with male C57BL/6 mice, then trophoblastic cells were obtained and its antigen was extracted. After co-cultured with TbC-Ag or spleen cells antigen of C57BL/6 mice (SC-Ag), aaDC of Balb/c mice became into TbC-Ag aaDC or SC-Ag aaDC. Concentrations of IL-10 and IL-12 in the culture and expression of CD40, CD80, CD86 and MHC-Ⅱ by aaDC loaded with antigen were detected. At 7th day after Balb/c recipients were intravenously injected with TbC-Ag aaDC (TbC-Ag group) or SC-Ag aaDC (SC-Ag group) or normal saline (control group), heterotopic vascularized heart transplantation was performed from C57BL/6 to BALB/c mice. As third donor group, heterotopic vascularized heart transplantation was performed from Kunming mice to BALB/c mice injected with TbC-Ag aaDC. The cardiac allograft survival time was compared. The expression of of IL-2, IL-10 and IFN-γ, mRNA in heart grafts and recipient spleen cells was detected by RT-PCR. Results The expression of CD86 and MHC-Ⅱ by SC-Ag aaDC was elevated, while only expression of MHC-Ⅱ by TbC-Ag aaDC was elevated. In the culture of TC-Ag aaDC, concentrations of IL-10 and IL-12 were (122.6±15.4) and (232.8±25.4) ng/L, while those of control group were (35.4±8.5) and (267.3±12.5) ng/L, respectively. There was significant difference in the IL-10 concentration (P<0.05), but no significant difference in IL-12 (P>0.05) between two groups. There were no significant differences in IL-10 and IL-12 concentrations between SC-Ag aaDC and blank control groups (P>0.05). The cardiac allograft survival time was (6.73± 0.58) days in control group, (12.28±0.76) days in SC-Ag group, (38.83±2.79) days in TbC-Ag group, and (6.68±0.62) days in third donor group, respectively. The survival time in TbC-Ag group was significantly longer than that in control and SC-Ag groups (P<0.01), but there was no significant difference in survival time between control and third donor groups (P>0.05). Conclusion Infusion of TbC-Ag aaDC results in prolonged survival of a completely MHC-mismatched heart allograft.