Yarchoan R, Murphy B R, Strober W, Schneider H S, Nelson D L
J Immunol. 1981 Dec;127(6):2588-94.
Cultures of human peripheral blood mononuclear cells stimulated in vitro with influenza virus were found to produce anti-influenza virus antibody. An enzyme-linked immunosorbent assay was used to measure cumulative antibody secretion into culture supernatants. Optimal antibody production was obtained with 2 x 10(6) peripheral blood mononuclear cells cultured in flat-bottomed wells for 12 days in 2 ml of RPMI 1640 medium supplemented with 10% fetal calf serum, antibiotics, and 4 mM L-glutamine. Antibody could be detected in cultures stimulated with live A/Hong Kong/68 (H3N2), purified formalin-inactivated A/Aichi/68 (H3N2), or live B/Hong Kong/68 influenza virus. Antibody of both IgG and IgM isotypes was produced. Under optimum conditions, the secreted IgG antibody was found to be directed preferentially at antigens on the stimulating virus. Antibody production was shown to require the cooperative interaction of B and T cells. Poisson analysis of B cell precursor frequency showed that approximately 1 in 40,000 B cells could be induced to produce IgG anti-influenza virus antibody in virus-stimulated cultures. Further investigation of this in vitro response should allow dissection of the cellular mechanisms involved in the regulation of antibody production in man.
研究发现，用流感病毒在体外刺激人外周血单个核细胞培养物可产生抗流感病毒抗体。采用酶联免疫吸附测定法来测量培养上清液中累积的抗体分泌量。在补充有10%胎牛血清、抗生素和4 mM L-谷氨酰胺的2 ml RPMI 1640培养基中，将2×10⁶外周血单个核细胞在平底孔中培养12天，可获得最佳抗体产量。在用活的A/香港/68（H3N2）、纯化的福尔马林灭活的A/爱知/68（H3N2）或活的B/香港/68流感病毒刺激的培养物中可检测到抗体。产生了IgG和IgM两种亚型的抗体。在最佳条件下，发现分泌的IgG抗体优先针对刺激病毒上的抗原。结果表明，抗体产生需要B细胞和T细胞的协同相互作用。对B细胞前体频率的泊松分析表明，在病毒刺激的培养物中，大约每40000个B细胞中有1个可被诱导产生IgG抗流感病毒抗体。对这种体外反应的进一步研究应有助于剖析人体中抗体产生调节所涉及的细胞机制。
