细胞因子诱导的imDCs鉴定及免疫学功能研究  

Immunological function and identification of imDCs induced by cytokine

作　　者：杨麦贵[1] 杨阳[2] 岳波[3] 郭树忠[4] 庄然[5] 张小宁[1] 郝晓柯[1] 

机构地区：[1]第四军医大学西京医院全军临床检验医学中心,西安市710032 [2]中国人民解放军第252医院整形科,河北省保定市071051 [3]第四军医大学西京医院全军耳鼻咽喉头颈外科中心,西安市710032 [4]第四军医大学西京医院全军整形外科研究所,西安市710032 [5]第四军医大学基础部免疫学教研室,西安市710032

出　　处：《中华实用诊断与治疗杂志》2011年第5期466-469,共4页Journal of Chinese Practical Diagnosis and Therapy

基　　金：国家自然科学基金重点项目(30830102);国家自然科学基金面上项目(81071581)

摘　　要：目的:分析未成熟树突状细胞(immature dendritic cells,imDCs)的免疫学功能,诱导同种异体复合组织移植局部免疫耐受性。方法:采用细胞因子诱导培养黏附法,取大鼠骨髓前体细胞,粒细胞-巨噬细胞集落刺激因子联合白细胞介素-4细胞因子诱导下培养7 d,收集贴壁细胞光学显微镜观察imDCs的形态学特征,用质量分数2%锥虫蓝染色检测活性细胞率;应用流式细胞仪分析imDCs表面标志分子;MTT比色法测定imDCs与同种异体T淋巴细胞混合培养后T淋巴细胞的增殖能力。结果:制备诱导培养的imDCs具有形态不规则、表面粗糙层叠状皱褶、胞质突起短而多、树枝样分叉明显等典型形态特征;活性细胞率高达93%;表面较特异标志分子OX62为(61.08±4.86)%,OX62阳性细胞表达CD80、CD86、组织相容性复合体-Ⅱ表达率分别为(3.56±0.73)%、(5.38±1.09)%、(6.45±0.74)%,均为较低表达;imDCs对同种异体T淋巴细胞基本无刺激增殖能力。结论:用细胞因子诱导培养黏附法取骨髓前体细胞,可较快分选出活性和纯度较高、数量足够的imDCs,为同种异体复合组织移植诱导免疫耐受防止排斥反应的研究奠定良好的基础。Objective To analyze the immunological function of immmature dendritic cells and research local immunologic tolerance of composite tissue allotransplantation induced by immature dendritic cells.Methods The bone marrow precursor cells were cultured for 7 days in rats induced by granular-macrophage colony stimulating factor and interleukin-4.The adherent cells were collected to study the morphological characteristics of immature dendritic cells under optical microscope,scanning microscope and transmission microscope.The activity rate of cell was detected with 2% trypan blue.The marker molecule of immature dendritic cells was analyzed with flow cytometry.T cells＇ proliferation potential was detected with MTT after immature dendritic cells were cultured and mixed with allograft T lymphocyte cells.Results Immature dendritic cells exhibited typical morphological characteristics,such as irregular shapes,rough surface with multi-layer ruga,various short cytoplasmic processes and obvious arboreal bifurcations.The viable cells accounted for 93%.The expression rates of surface specific marker molecular OX62,CD80,CD86,MHC-Ⅱ were（61.08±4.86）%,（3.56±0.73）%,（5.38±1.09）%,（6.45±0.74）% respectively,and they all lowly expressed.Immature dendritic cells almost had no stimulation on the proliferation of allogenic T lymphocytes.Conclusion Bone marrow precursor cells can be separated out enough immature dendritic cells with a high activity and purity,which provides a good foundation for preventing immunologic tolerance rejection reaction induced by composite tissue allotransplantation.

关 键 词：同种异体复合组织移植 细胞因子 骨髓 未成熟树突状细胞 局部免疫耐受 

分 类 号：R392.4[医药卫生—免疫学]