CCR7基因修饰未成熟树突状细胞诱导小鼠皮肤移植免疫耐受的研究

Research on induction of immune tolerance to murine skin allograft by CCR7 gene modified immature dendritic cells

目的研究CCR7基因重组腺病毒体外转染未成熟树突状细胞(imDCs)对小鼠同种异体皮肤移植免疫耐受的影响。方法小鼠骨髓细胞经梯度离心及rmIL-4、rmGM-CSF诱导培养未成熟树突状细胞;CCR7重组腺病毒经增强离心法转染未成熟树突状细胞;体外观察转染细胞混合淋巴细胞反应;取CCR7修饰imDCs于皮肤移植前2d、移植后第5天,经腹部注入受体小鼠,观察皮肤移植物存活时间和病理变化。结果CCR7基因成功转染入imDCs;腺病毒转染后imDCs刺激T细胞增殖的能力较imDCs组增强但弱于mDCs组,IL-10可以明显降低刺激能力;CCR7腺病毒转染组皮肤存活时间长于转染前,IL-10能显著延长皮肤存活时间。结论CCR7重组腺病毒转染imDCs后,imDCs刺激T细胞增殖的能力部分增强,皮肤移植物存活时间明显延长。

Objective To investigate the influence of immune tolerance to murine skin allograft transfected with CCR7 recombnant adenovirus in vitro. Methods Immature dendritic cells generated in the presence of recombinant murine interleukin-4 and granuloeyte/macrophage colony-stimulating factor from murine bone marrow by gradient eentrifugation method; CCR7 recombinant adenovirus vector was transfected to immature dendritic cells by centrifugal force method; The mixed lymphocyte reaction of imDCs were observed after transfected with CCR7 recombinant adenovirus; The imDCs modified by CCR7 gene were infused into receptor routine through abdominal cavity before 2 days and after 5 days of graft, the MST and pathological change of skin graft were observed. Results The CCR7 gene had successfully transfected into imDCs; The ability of stimulate T cell proliferation of imDCs after transfection was enhanced but lower than group mDCs, IL-10 could descended the stimulate ability obviously; The MST of CCR7 transfeeted group was longer than untransfected group,IL-10 could significant prolongated the MST of skin graft. Conclusion The imDCs transfects with CCR7 recombinant adenovirus,the ability of stimulate T cell proliferation improves slightly,the MST of skin graft prolongated obviously.