人食管癌Eca-109细胞膜抗原筛选的初步研究

The preliminary study on screening cytomembrane antigens in human esophageal carcinoma Eca-109 cell line

目的 从人食管癌Eca 10 9细胞入手 ,寻找能引起CTL免疫反应的肿瘤抗原 ,筛选出高效肿瘤抗原的大致范围。方法 采用敏感的序列特异性引物 聚合酶链反应 (PCR SSP)来确定Eca 10 9细胞HLA基因分型 ,用HLA血清学分型技术 (Terasaki改良的微量细胞毒试验 )筛选健康志愿者 ;用MTT法测定致敏T细胞的细胞毒活性。结果 Eca 10 9细胞的HLA的基因分型为A 0 3 ,A 2 4;B 3 5 ,B 3 7;DRB1 0 1,DRB1 12 ;DRB3 ;筛选到一例表型为A0 3杂合子的健康志愿者。根据MTT检测 ,Eca 10 9细胞小于 10KD膜蛋白免疫原性较未使用去垢剂的总膜蛋白的免疫原性高 ,小于 3KD膜蛋白的免疫原性最高。结论 Eca 10 9细胞膜蛋白中存在能引起特异性的抗肿瘤CTL免疫反应的肿瘤抗原 ,其中小于

Objective To search and screen out tumor antigens and the rough scope of high performance tumor antigens of Eca-109 cell line that can stimulate cytolytic T lymphocyte to produce anti-tumor immune response.Methods For HLA genotyping, the sequence specific primer polymerase chain reaction(PCR-SSP) method used allele-specific primers in the amplifications of PCR. The HLA alleles of Eca-109 cell were identified. With serological HLA typing technology, the healthy adult volunteers' HLA alleles were screened to match at least one of the HLA alleles of Eca-109 cell. By MTT reduction assay, specific cytotoxicity of CTL against Eca-109 cells was analyzed.Results The HLA alleles of Eca-109 were A *03, A *24; B *35, B *37;DRB1 *01,DRB1 *12;DRB3, and a healthy volunteer with heterozygote HLA-A03 was found. In MTT reduction assay, the immunogenicity of less than 10KD cytomembrane proteins of Eca-109 cell was more efficient than that of other cytomembrane proteins with no detergent, and the immunogenicity of the proteins less than 3KD was the most effective in the experimental groups. Conclusion In cytomembrane proteins of Eca-109 cell, there were tumor antigens that could stimulate antitumor immune response and the cytomembrane proteins less than 3KD may have tumor antigens with high efficiency against human esophageal carcinoma.