Maslak D M, Reynolds D L
Department of Microbiology, Immunology, and Preventive Medicine, Iowa State University, Ames, Iowa 50011, USA.
Avian Dis. 1995 Jan-Mar;39(1):1-8.
A blastogenesis microassay employing 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) was adapted to measure blastogenic responses of lymphocytes from the chicken's head-associated lymphoid tissues (i.e., the harderian gland and conjunctiva-associated lymphoid tissue) to T- and B-cell mitogens. Lymphocytes isolated from peripheral blood, spleen, and the harderian gland had highly significant (P < 0.01) responses to T- and B-cell mitogens compared with control lymphocytes cultured without mitogens. Cultured lymphocytes obtained from the harderian gland had highly significant mitogenic responses to the T-cell mitogen concanavalin A (25 to 100 micrograms/ml) and to the B-cell mitogen Salmonella typhimurium lipopolysaccharide (1.25 to 5.0 micrograms/ml) compared with the control lymphocytes. Mitogenic responses of cultured lymphocytes obtained from the conjunctiva-associated lymphoid tissue could not be measured within the given parameters of the blastogenesis microassay. This was primarily due to the low yield of lymphocytes, which proved to be a limiting factor. The ability of the MTT blastogenesis microassay to detect blastogenic responses of the harderian gland to mitogens may be indicative of its usefulness for measuring cell-mediated immunity responses to other antigens.
采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)的增殖反应微量测定法,用于检测鸡头部相关淋巴组织(即哈德氏腺和结膜相关淋巴组织)中淋巴细胞对T细胞和B细胞有丝分裂原的增殖反应。与未添加有丝分裂原培养的对照淋巴细胞相比,从外周血、脾脏和哈德氏腺分离的淋巴细胞对T细胞和B细胞有丝分裂原具有高度显著(P < 0.01)的反应。与对照淋巴细胞相比,从哈德氏腺获得的培养淋巴细胞对T细胞有丝分裂原刀豆球蛋白A(25至100微克/毫升)和B细胞有丝分裂原鼠伤寒沙门氏菌脂多糖(1.25至5.0微克/毫升)具有高度显著的促有丝分裂反应。在增殖反应微量测定法给定的参数范围内,无法检测从结膜相关淋巴组织获得的培养淋巴细胞的促有丝分裂反应。这主要是由于淋巴细胞产量低,这被证明是一个限制因素。MTT增殖反应微量测定法检测哈德氏腺对有丝分裂原的增殖反应的能力,可能表明其可用于测量对其他抗原的细胞介导免疫反应。