胚胎干细胞源性树突状细胞对同源性神经前体细胞脑内移植耐受诱导  

Dendritic cells differentiated from 129/svj ES-D3 embryonic stem cells prolong syngeneic neural progenitor cells survival in ischemic rat brain

作　　者：梅爱农[1] 张苏明(指导)[1] 

机构地区：[1]福建医科大学省立临床医学院干部特诊一科,福建省临床老年病研究所,福建省立医院老年医学研究室

出　　处：《中国免疫学杂志》2015年第3期318-322,328,共6页Chinese Journal of Immunology

摘　　要：目的:观察脑缺血环境下胚胎干细胞源性树突状细胞(es DCs)能否诱导同源性神经前体细胞(NPCs)的移植耐受。方法:分别自129/svj p CX-e GFP ES-D3诱生NPCs及129/svj ES-D3诱生es DCs。线栓法制作MCAo模型SD大鼠并相应分为预处理组及对照组,术后1周预处理组经尾静脉注射es DCs而对照组注射PBS,两组动物在预处理1周后侧脑室注射NPCs,2周后(MCAo术后4周)免疫组化观察纹状区CD4+、CD8+、ED1+细胞的分布差异,MTT法观察颈部淋巴细胞增殖性(PI),逆转录PCR(RT-PCR)半定量局部IL-10、IFN-γmRNA表达差异,荧光显微镜观察GFP标记NPCs海马存活率。结果:预处理组大鼠,CD4+细胞浸润显著低于对照组(134.7±36.2 vs 198.8±59.6,P<0.05),NPCs存活率显著高于对照组(P<0.05),但预处理组对ED1+(298.8±75.9 vs 302.2±88.5)、CD8+(145.8±45.4 vs 134.3±39.0)细胞浸润、局部细胞因子mRNA IFN-γ(1.697±0.273 vs 1.829±0.250)、IL-10(1.147±0.260 vs 1.264±0.119)及外周淋巴细胞增长率(PI,1.245±0.211 vs1.331±0.235)无明显影响。结论:es DC可能通过降低局部CD4+细胞反应诱导针对同源性NPCs免疫耐受,尚无确切证据表明细胞因子途径参与该过程,确切的机理尚需更深入研究。Objective：To study if embryonic stem cell derived dendritic cells （esDCs） could induce transplant tolerance to syngeneic neural progenitor cells （NPCs） in ischemic rat brain. Methods： Neural progenitor cells （NPCs） were differentiated from 129/svj pCX-eGFP ES-D3 embryonic stem cells and dendritic ceils were directly differentiated from 129/svj ES-D3 respectively. All of SD rats were accepted MCAo surgery and subdivided in two groups based on pretreatment with or without esDCs through tail vein injection 1 week after MCAo. pCX-eGFP NPCs were then injected into the lateral ventricle of animals 2 weeks after MCAo. A proliferation assay of lymphocytes dissociated from cervical lymph nodes by MTr method, counting of the survival of the grafted cells, histological evaluation of CD4, CD8 and ED1 positive cells in brain and detection of mRNA level of IL-10 and IFN-Y in ischemic lesions by reverse transcriptase-polymerase chain reaction（RT-PCR） were performed 2 weeks after graft （4 weeks after MCAo）. Results：Pre- treatment with esDCs decreased CD4 positive cells infiltration （ 134. 7 ± 36. 2 vs. 198. 8 ± 59. 6 ,P〈0. 05） and enhance the survival of NPCs in ischemic striatum,but had no effect on ED1 （298. 8 ± 75.9 vs. 302. 2＋88.5） ,CD8 positive cells （ 145.8 ±45.4 vs. 134. 3± 39. 0） distribution （P〉O. 05）. There was also no difference in lymphocytes proliferation （ PI, 1. 245 ± 0. 211 vs. 1.331 ± 0. 235 ） or mRNA expression level of IL-IO （1. 147±0.260 vs. 1.264±0. 119） and IFN-Y （1.697 ± 0.273 vs. 1.829 ±0.250） between two groups （ P〉0. 05 ）. Conclusion ： The results indicate that pretreatment with esDCs may prolong syngeneic NPCs survival though reducing CD4 positive cells reaction in ischemic striatum,which provides some evidence for the tolerogenic function of esDCs. However,there was lack of evidence for cytokine-dependent routine involving in this mode and further investigation was needed to make certain the cardinal principle.

关 键 词：胚胎干细胞 神经前体细胞 树突状细胞 移植耐受 脑缺血 

分 类 号：R392.4[医药卫生—免疫学]