Quade M J, Roth J A
Department of Veterinary Microbiology, and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames 50011, USA.
Vet Immunol Immunopathol. 1999 Jan 4;67(1):33-45. doi: 10.1016/s0165-2427(98)00209-8.
Bovine peripheral blood mononuclear cells were cultured in vitro for 3 days with the mitogens concanavalin A (Con A), pokeweed mitogen (PWM), phytohemagglutinin (PHA), and anti-CD3 monoclonal antibody. Activation of T-lymphocyte subsets (CD4+, CD8+, and gammadelta T-cell receptor+) and of B-cells was measured by two-color flow cytometric analysis of subset expression of IL-2 receptor alpha (CD25) and MHC class II. Proliferation of lymphocyte subsets was directly measured by two-color flow cytometric analysis of fluorescence intensity of PKH2, a fluorescent dye that stably incorporates into cell membranes. CD4+ and CD8+ T-cell subsets were stimulated by all the mitogens to increase expression of IL2r and MHC II and to proliferate. Deltagamma+ T-cells responded to all four mitogens with increased IL2r and MHC II expression. Con A and PHA caused measurable proliferation of deltagamma+ T-cells, but PWM and anti-CD3 did not. B-cells generally responded to the mitogens with increased IL2r and MHC II expression. B-cells proliferated when incubated with Con A, but did not measurably proliferate in response to PWM, PHA, or anti-CD3.
牛外周血单个核细胞与丝裂原刀豆球蛋白A（Con A）、商陆丝裂原（PWM）、植物血凝素（PHA）和抗CD3单克隆抗体在体外培养3天。通过对白细胞介素2受体α（CD25）和MHC II类分子亚群表达进行双色流式细胞术分析，检测T淋巴细胞亚群（CD4 +、CD8 +和γδT细胞受体+）以及B细胞的活化情况。通过对稳定掺入细胞膜的荧光染料PKH2的荧光强度进行双色流式细胞术分析，直接检测淋巴细胞亚群的增殖情况。所有丝裂原均刺激CD4 +和CD8 + T细胞亚群，使其白细胞介素2受体（IL2r）和MHC II表达增加并发生增殖。γδ + T细胞对所有四种丝裂原均有反应，白细胞介素2受体（IL2r）和MHC II表达增加。Con A和PHA可使γδ + T细胞发生可测量的增殖，但PWM和抗CD3则不能。B细胞通常对丝裂原有反应，白细胞介素2受体（IL2r）和MHC II表达增加。B细胞与Con A孵育时会发生增殖，但对PWM、PHA或抗CD3则无明显增殖反应。
