中文摘要：

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’CDl9＋调节性B细胞（Breg）比例及移植物存活时间的影响。方法以C57BL／6d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／b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（sv），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／6S小鼠骨髓来源的imDC共同培养，得NPUVA．SPDCs。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．SPDC组、imDC组、成熟树突细胞（maDC）组和PBS对照组。于手术前7d分别从尾静脉注人1×10。个（0．2m1）PUVA．SPDC、imDC、maDC或0．2mlPBS。于移植术后观察受体小鼠的移植物存活时间、外周血IL-10’CDl9’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’CDl9’Breg细胞占CDl9’B细胞的比例在PUVA．SPDC组为7．48％，明显高于imDC组（4．12％）、maDC组（3．ol％）和PBS对照组（2．37％）。PUVA．SPDC组小鼠血清中IL．10表达水平为58．2±o．9ng／ml，与maDC组（20．1±1．6ng／m1）、imDC组（26．24-1．3ng／m1）及PBS对照组（19．0±o．6ng／m1）比较显著升高（P〈0．01）。PUVA．SPDC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．SPDCs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。

英文摘要：

Objective To investigate the effect of recipient immature dendritic cells （imDCs） loaded with PUVA-treated donor apoptotic splenic lymphocytes （PUVA-SP DC） on IL-10＋CD19＋regulatory B cells （Breg） and the survival duration of skin allograft in mice. Methods Bone marrow-derived DCs of CS7BL/6 mice were obtained from bone marrow cells by co-culturing with recombinant mouse IL-4 and GM-CSF. Spleen lymphocytes （SP） of BALB/c mice were isolated and prepared as PUVA- SP by treating the cells with 8-methoxypsoralen plus ultraviolet A irradiation. The bone marrow-derived imDCs of CS7BL/6 mice were co-cultured with PUVA-SP of BALB/c mice to obtain PUVA-SP DCs. The skin allograft model was then established. Animals were randomly grouped according to different pretreatments as follows： the control group was iv. introduction of PBS （0.2ml） alone 7 days before skin transplantation, the PUVA-SP DC group received an iv. injection of PUVA-SP DCs, the maDC （mature DC） group received recipient maDCs, and the imDC group was given recipient imDCs. Mice were monitored daily from day 6 after transplantation for signs of rejection of skin graft. The recipients＇ peripheral blood serum samples were then collected and the level of cytokines were measured by using ELISA kits. The survival time of skin allograft was evaluated every day. The expression of IL-10＋CD19＋regulatory B cells was analyzed by flow cytometry. Results After transplantation, the proportion of IL-10＋CD19±Breg in the peripheral blood of PUVA-SP DC group was 7.48%, which was obviously higher than that of imDC group （4.12%）, maDC group （3.01%） and control group （2.37%）. The serum level of cytokine IL-10 in PUVA-SP DC group was 58.2 ＋ 0.9ng/ml, and it was significantly higher than that in maDC group （20.1 ± 1.6ng/ml）, imDC group （26.2 ± 1.3ng/ml） and control group （19.0 ± 0.6ng/ ml, P〈0.01）. The survival time of allograft in PUVA-SP DC group was 62.3 ＋ 2.6d, and it was markedly longer than that in maDC gr