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目的探讨吞噬了供者凋亡淋巴细胞的未成熟树突细胞(imDC)对皮肤移植受体小鼠外周血IL-10+CD19+调节性B细胞(Breg)比例及移植物存活时间的影响。方法以C57BL/6小鼠作为受者,BALB/c小鼠为供者,建立小鼠皮肤移植模型。分离C57BL/6小鼠骨髓细胞,经小鼠重组白细胞介素4(IL-4)和粒-巨噬细胞集落刺激因子(GM-CSF)共同诱导,制备并培养imDC。分离BALB/c小鼠脾淋巴细胞(SP),经光化学照射方法(PUVA)处理,得到供者小鼠脾淋巴细胞(PUVA-SP)。在体外将PUVA-SP与C57BL/6小鼠骨髓来源的imDC共同培养,得到PUVA-SP DCs。根据受体小鼠术前接受的静脉输注成分将其随机分为4组(n=14):PUVA-SP DC组、imDC组、成熟树突细胞(maDC)组和PBS对照组。于手术前7d分别从尾静脉注入1×l06个(0.2ml)PUVA-SP DC、imDC、maDC或0.2ml PBS。于移植术后观察受体小鼠的移植物存活时间、外周血IL-10+CD19+Breg比例及IL-10的表达情况。结果移植术后,受体小鼠外周血IL-10+CD19+Breg细胞占CD19+B细胞的比例在PUVA-SP DC组为7.48%,明显高于imDC组(4.12%)、maDC组(3.01%)和PBS对照组(2.37%)。PUVA-SP DC组小鼠血清中IL-10表达水平为58.2±0.9ng/ml,与maDC组(20.1±1.6ng/ml)、imDC组(26.2±1.3ng/ml)及PBS对照组(19.0±0.6ng/ml)比较显著升高(P<0.01)。PUVA-SP DC组移植物存活时间为62.3±2.6d,显著长于maDC组(20.7±1.9d)、imDC组(12.1±1.0d)和PBS对照组(11.0±1.3d,P<0.01)。结论移植术前输注PUVA-SP DCs可显著延长移植物存活时间,增加受者体内IL-10的表达水平,诱导产生较多分泌IL-10的调节性B细胞。
Objective To investigate the effects of imDC phagocytes (imDC) engulfing donor apoptotic lymphocytes on the ratio of IL-10 + CD19 + regulatory B cells (Breg) and graft survival in peripheral blood of skin graft recipients. Methods C57BL / 6 mice were used as recipients and BALB / c mice as donors to establish mouse skin transplantation model. Bone marrow cells of C57BL / 6 mice were isolated and induced by mouse recombinant interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF) to prepare and culture imDC. Splenic lymphocytes (SPs) of BALB / c mice were isolated and treated with PUVA to obtain splenic lymphocytes (PUVA-SP) from donor mice. PUVA-SP was co-cultured with imDC derived from C57BL / 6 mouse bone marrow in vitro to obtain PUVA-SP DCs. The recipient mice were randomly divided into 4 groups (n = 14): PUVA-SP DC group, imDC group, mature DC group and PBS control group, according to the preoperative intravenous infusion component of the recipient mice. One hundred and sixty (0.2 ml) PUVA-SP DC, imDC, maDC, or 0.2 ml PBS were injected from the tail vein 7 days prior to surgery. The graft survival time, the ratio of IL-10 + CD19 + Breg in peripheral blood and the expression of IL-10 in recipient mice were observed after transplantation. Results After transplantation, the ratio of IL-10 + CD19 + Breg cells to CD19 + B cells in peripheral blood of recipient mice was 7.48% in PUVA-SP DC group, significantly higher than that in imDC group (4.12%), maDC group %) And PBS control group (2.37%). The level of IL-10 in serum of PUVA-SP DC group was 58.2 ± 0.9ng / ml, which was significantly higher than that of maDC group (20.1 ± 1.6ng / ml), imDC group (26.2 ± 1.3ng / ml) and PBS control group ± 0.6ng / ml) was significantly higher (P <0.01). PUVA-SP DC group graft survival time was 62.3 ± 2.6d, significantly longer than the maDC group (20.7 ± 1.9d), imDC group (12.1 ± 1.0d) and PBS control group (11.0 ± 1.3d, P <0.01). Conclusions Transplantation of PUVA-SP DCs before transplantation can significantly prolong graft survival, increase IL-10 expression in recipients and induce more B-regulatory cells that secrete more IL-10.