混合培养的供、受者骨髓细胞过继回输延长小鼠移植心脏存活时间

目的 将供、受者骨髓细胞经混合培养后过继回输,以观察其对同种异体移植心脏存活时间和受者免疫功能的影响.方法 取Balb/c小鼠和C57BL/6J小鼠的骨髓细胞,进行混合培养.配制含Balb/c小鼠和C57BL/6J小鼠脾淋巴细胞的混合淋巴细胞反应体系(MLR)以及含Balb/c小鼠和C3H小鼠脾淋巴细胞的MLR,分别加入混合培养的骨髓细胞,观察其对MLR中细胞增殖的影响.以C57BL/6J小鼠为供者,Balb/c小鼠为受者行腹腔异位心脏移植,实验分为4组:(1)移植对照组,受者仅进行心脏移植,不作其他处理;(2)实验对照组,心脏移植后给予西罗莫司灌胃;(3)实验组,移植手术结束前注射混合培养的骨髓细胞1×10~7个,术后给予西罗莫司;(4)第三方对照组,受者接受C3H小鼠的移植心脏,手术结束前注射混合培养的骨髓细胞1×10~7个,术后给予西罗莫司.记录移植心脏存活时间;移植心脏停跳当日,取受者外周血,检测CD4~+ CD25~+ T淋巴细胞的比例及供者来源的H-2K~b细胞的比例.结果 加入混合培养的骨髓细胞后,Balb/c和C57BL/6J的MLR的淋巴细胞增殖率低于Balb/c和C3H的MLR.实验组移植心脏的存活时间长于其他3组(P<0.05).实验组CD4~+CD25~+T淋巴细胞的百分率高于其他3组(P<0.05).实验组外周血中H-2K~b细胞的比例高于其他3组(P<0.05).结论 受者输注混合培养的供、受者骨髓细胞可在一定程度上调节免疫应答,延长小鼠移植心脏的存活时间,该作用具有供者抗原特异性.

Adoptive transfer of mix-cultured hone marrow cells to prolong survival of cardiac allografts in mice

Objective To investigate the effects of adoptive transfer of mix-cultured bone marrow cells (BMC) from donor and recipient on allograft and immune system after the recipients preconditioned by a non-myeloablative regimen.Methods Mix-cultured BMC were added into mixed lymphocyte reaction.The effects of mix-cultured BMC on lymphocyte proliferation and their specificity were observed.Heart from C57BL/6 mouse was transplanted into Balb/c abdominal cavity,and the BMC from donor and recipient was mix-cultured and adoptively transferred.The recipients were divided as follows:Ⅰ,no treatment;Ⅱ,total body irradiation (TBI) + rapamune;Ⅲ,TBI + rapamune + mix-cultured BMC;Ⅳ,treated by the same protocol as group Ⅲ except that C3H mouse served as the donor.The survival of cardiac allograft in all groups was observed.The percentage of CD4~+ CD25~+ T and donor-derived H-2K~b cells in peripheral blood of the recipient was measured by using FCM,when the allograft stopped beating.Results Mix-cultured BMC specifically downregulated lymphocyte reaction in vitro.Adoptive transfer of mix-cultured BMC significantly prolonged the allograft survival in group Ⅲ,but not in group Ⅳ.And in recipient peripheral blood of group Ⅲ,the percentage of CD4~+ CD25~+ T cells and H-2K~b cells was increased significantly.Specially,these effects were all acted as a donor antigen-specific manner.Conclusion Mix-cultured BMC can regulate immune response and prolong the cardiac allograft survival in a donor antigen-specific manner.