摘要
目的制备人工抗原提呈细胞(artificial antigen presenting cell,aAPC)并用它从HLA-A2阳性健康个体外周血单个核细胞(PBMC)中诱导和扩增特异性细胞毒性T淋巴细胞(CTL)。方法将HLA-A2-EBV四聚体分子和抗CD28抗体分子吸附固定在细胞大小的聚苯乙烯乳胶微球(5μm)表面制成aAPC；采用流式细胞仪表型分析；aAPC和HLA-A2阳性个体人外周血单核细胞进行混合淋巴细胞反应；用HLA-A*0201-EBV四聚体染色法检测特异性CTL的频率；应用细胞内细胞因子染色法检测特异性CTL功能性细胞因子IFN-γ的分泌；采用LDH释放法检测特异性CTL的特异杀伤活性。结果流式细胞仪分析显示微球表面吸附有HLA-A2-EBV四聚体分子和抗CD28抗体分子；四聚体检测及细胞内细胞因子染色法检测与经典细胞毒试验结果一致,结果表明aAPC在体外可诱导抗原特异性CTL的生成。结论aAPC制备成功,并在体外有效地诱导抗原特异性CTL的生成。
Objective To prepare artificial antigen presenting cells （aAPC） and to induce specific cytotoxic T cells （CTL） using aAPC from peripheral blood mononuclear cell （PBMC） from HLA-A2＊ normal donors. Methods aAPC was generated by coupling HLA-A2-EBV and anti-CD28 antibody to the surface of the cell-size microbeads. The immunophenotype of aAPC analyzed by FACS. PBMC from HLA-A2^＋ donors were co-cultured with aAPCs. Specific CTL response was demonstrated by HLA-A2-EBV tetramer staining, intracellular cytokine stain assay and cytotoxicity assay. Results The analysis of FACS shows that HLA-A2-EBV and anti-CD28 antibody are immobilized on microdeadds. The result of tetramer staining, intracellular cytokine stain assay and cytotoxicity assay show that aAPC successfully induces specific CTL successfully. Conclusion aAPC was prepared and could induce specific CTL in vitro.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2006年第10期871-874,共4页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金(30271201)
科技部973计划资助(2001CB510008)
