Wang C Y, Al-Katib A, Lane C L, Koziner B, Fu S M
J Exp Med. 1983 Nov 1;158(5):1757-62. doi: 10.1084/jem.158.5.1757.
The expression of HLA-DC/DS antigen detected by the monoclonal antibody Leu 10 was studied in three human precursor and pre-B cell lines (Josh 7, Reh, and Nalm 12). Flow cytometric analysis showed that none of these cell lines stained for the HLA-DC/DS antigen. In the presence of 1.6 X 10(-9) M of 12-O-tetradecanoylporbol-13-acetate (TPA), expression of this antigen was detected. The expression was completed after 168 h of incubation. Iodination of cell surface, immunoprecipitation by Leu 10 antibody, and two-dimensional gel analysis revealed that TPA-treated Josh 7 cells synthesized and expressed a 29,34 kD bimolecular complex with both alpha and beta chains different from those of HLA-DR antigen. Quantitative absorption experiments with cell lysates indicated a greater than 25-fold increase in HLA-DC/DS antigen in TPA-treated cells. With the induction of HLA-DC/DS antigen expression, there are concomitant decreases in the expression of the common acute lymphoblastic leukemia antigen (CALLA) and the enzymatic activity of terminal deoxynucleotidyl transferase. No appreciable changes in HLA-DR and Ig expression were observed. There was also no change in HLA-SB expression as detected by antibody ILR-1. However, DNA synthesis was markedly inhibited by TPA treatment. These results indicate that precursor and pre-B cell lines can be induced to mature in vitro. They also suggest that the expression of HLA-DC/DS antigen which precedes the expression of membrane Ig and follows the HLA-DR expression is relevant to human B cell development and cell interaction.
用单克隆抗体Leu 10检测HLA - DC/DS抗原在三个人类前体和前B细胞系(Josh 7、Reh和Nalm 12)中的表达情况。流式细胞术分析显示,这些细胞系均未检测到HLA - DC/DS抗原染色。在存在1.6×10⁻⁹ M的12 - O - 十四烷酰佛波醇 - 13 - 乙酸酯(TPA)时,可检测到该抗原的表达。孵育168小时后表达完成。细胞表面碘化、Leu 10抗体免疫沉淀及二维凝胶分析表明,经TPA处理的Josh 7细胞合成并表达了一种29.34 kD的双分子复合物,其α链和β链均与HLA - DR抗原不同。用细胞裂解物进行的定量吸收实验表明,经TPA处理的细胞中HLA - DC/DS抗原增加了25倍以上。随着HLA - DC/DS抗原表达的诱导,常见急性淋巴细胞白血病抗原(CALLA)的表达及末端脱氧核苷酸转移酶的酶活性随之降低。未观察到HLA - DR和Ig表达有明显变化。用抗体ILR - 1检测时,HLA - SB表达也无变化。然而,TPA处理显著抑制了DNA合成。这些结果表明前体和前B细胞系可在体外被诱导成熟。它们还提示,在膜Ig表达之前且在HLA - DR表达之后的HLA - DC/DS抗原表达与人类B细胞发育及细胞相互作用相关。