表达特定免疫球蛋白可变区抗原的人B细胞系建立和其生物学特征

目的建立表达特定免疫球蛋白可变区抗原的人B细胞系。方法在29例EBV转化B淋巴细胞建立永生化淋巴母细胞系中,PCR-SSP方法检测HLA-A*0201表达的细胞系,将表达HLA-A*0201的细胞系通过有限稀释培养方法获得单克隆细胞系ZP-1。分别用PCR方法、流式细胞仪和染色体核型分析检测细胞IgHV家族、细胞表面免疫表型CD分子和染色体核型表达。结果 HLA-A*0201(+)的细胞,经单克隆化培养后获得3个细胞克隆,将其中之一命名为ZP-1。PCR检测显示ZP-1细胞表达IgHV3家族;流式细胞仪检测显示ZP-1细胞表达CD19、lambda链等B细胞免疫表型;核型分析结果显示,ZP-1细胞为正常女性染色体结构。结论成功获得表达特定免疫球蛋白可变区抗原的人B细胞系,为研发B细胞相关疾病的疫苗和分子靶向药物提供良好的实验基础。

Establishment and biological characteristics of human B cell line CZ-1 expressing specific immunoglobulin variable regions antigen

Objective Establishment of human B cell line expressing specific immunoglobulin variable regions antigen. Methods In the 29 cases of Epstein-Barr virus (EBV) transformed B lymphocytes to establish immortalized lymphoblastoid cell lines, PCR-SSP method detected the expression of HLA-A*0201. The HLA-A*0201(+) monoclonal cell line was obtained by limiting dilution culture, designated as ZP-1. The PCR, flow cytometry and chromosome analysis were used to detect the expression of IgHV family, phenotype CD molecules and chromosome, respectively. Results Three HLA-A*0201(+) cell clones were aquired by monoclone culture, one of these named ZP-1 cell. IgHV3 family was detected by PCR. The B cell phenotype was identified by flow cytometry. Such as CD19, lambda chain, et al. Karyotype analysis showed that ZP-1 cells have a normal female chromosome structure. Conclusions A human B cell line expressing specific immunoglobulin variable regions antigen was established. It provided a useful tool to develop the vaccine or molecular target drugs of B cell related diseases.