摘要
目的:建立猪外周血来源的未成熟树突状细胞(im-DC)体外诱导、扩增方法。探讨负载猪源性pCTLA4-Ig重组腺病毒的猪imDCs对大鼠脾细胞增殖的影响。方法:家猪外周血单个核细胞(PBMC),以rpGM-CSF(30μg/L)和rpIL-4(20μg/L)体外诱导培养,收集悬浮细胞经电镜、流式细胞仪鉴定。利用Adv-pCTLA4-Ig转染猪imDCs,RT-PCR鉴定。对比转染组与未转染组imDCs刺激SD大鼠脾细胞增殖的能力。结果:获得第4～5天悬浮细胞具有典型的imDCs形态特征,表达猪源性DC的特异性标志SLA-DR、CD172a,而CD80/CD86表达较低。Adv-pCTLA4-Ig转染猪imDCs行RT-PCR出现pCTLA4-Ig、吲哚胺2,3-二氧化酶(IDO)基因的表达。混合淋巴细胞培养转染组与未转染组imDCs刺激指数有显著性差异(P<0.05)。结论:联合应用rpGM-CSF和rpIL-4可诱导猪PBMC衍生成大量的imDCs。负载pCTLA4-Ig基因的猪imDCs,能够表达IDO,降低对异种抗原的免疫应答,显著抑制异种脾细胞增殖。
AIM：To establish a method for induced and amplified porcine monocyte-derived immature dendritic cells（imDCs）.The aim of this study was to assess the pCTLA4-Ig gene-modified porcine imDCs could benefit for prevent the proliferation of xenogeneic spleen cells.METHODS：Poricne peripheral blood mononuclear cells（PBMC） were cultured with rpGM-CSF（30 μg/L） and rpIL-4（20 μg/L）.To evaluated the suspending cells by transmission electronic microscope and flow cytometer.Porcine imDCs were transduced with adenovirus mediated pCTLA4-Ig gene.The mixed lymphocyte reaction（MLR） was used to examine the proliferation of spleen cells of SD rat to gene-modified imDCs.RESULTS：With rpGM-CSF and rpIL-4 induced porcine PBMC for 4-5 days,it was exhibited typical morphological characteristics and immunological phenotype of imDCs with high expression of SLA-DR,CD172a and low expression of CD80/CD86 on the cellular surface.The gene-modified imDCs were able to amplified the pCTLA4-Ig and IDO gene by RT-PCR.The stimulate index of gene-modified imDCs group were markedly inferior to that in unmodified imDCs group（P0.05）.CONCLUSION：The pCTLA4-Ig gene-modified imDCs can significantly inhibit the proliferation of xenogeneic spleen cells,and there might was correlated with the expression of IDO reduce the immune response of xenogeneic antigen.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2010年第5期434-437,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学青年基金资助项目(30700772)
