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

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

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）第三方对照纽，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“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H2K^6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。

Objective To investigate the effects of adoptive transfer of mix-cultured bone marrow ceils （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-2Kb ceils in peripheral blood of the recipient was measured by using FCM, when the allograft stopped heating. Results Mi^cultured BMC specifically down-regulated 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^6 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.