Wyckoff J H, Confer A W
Department of Veterinary Parasitology, College of Veterinary Medicine, Oklahoma State University, Stillwater 74078-0353.
Vet Immunol Immunopathol. 1990 Dec;26(4):367-83. doi: 10.1016/0165-2427(90)90120-h.
Regulation of the bovine immune response to immunization with Brucella abortus Strain 19 (S19) was investigated through application of a modification of an assay to measure suppressor T lymphocyte activities in humans and through development and characterization of antigen-stimulated T lymphocyte lines in vitro. A total of nine of steers were alloted into two groups: control (n = 4) and S19-immunized (n = 5). Peripheral blood mononuclear cells (PBMC) from each animal were cultured in vitro with mitogens (concanavalin A (Con A) and pokeweed mitogen (PWM], B. abortus antigens (B. abortus soluble antigen (BASA) and whole heat-killed B. abortus cells (HKC)) and media alone periodically from days 4 through 49 of the experiment. Supernates from these cultures were assayed for immunomodulatory activity(s) by addition to indicator cultures stimulated with suboptimal concentrations of Con A. Supernates from PBMC of S19-immunized steers generated with B. abortus antigens significantly (P less than 0.05) suppressed indicator cell responses as compared to those from control steers on days 35 and 49 post-immunization. This suppressive activity from PBMC of immunized cattle with respect to that of control cattle could also be induced through mitogenic stimulation with Con A or PWM. On day 49 of the study, suppressive activity was spontaneously released from the PBMC of immunized cattle. T lymphocyte lines were initiated from two S19-immunized steers at 2 and 9 weeks post-immunization. These T cell lines were characterized with respect to proliferative responses to B. abortus antigens through in vitro assay and surface marker expression through indirect immunofluorescence with a limited panel of monoclonal antibodies. Results from the present study indicated that S19 immunization induces a subpopulation(s) of cells in the PBMC of cattle capable of regulating the in vitro response to B. abortus. This regulatory activity is detectable by in vitro assay as early as 7 weeks post-immunization. Furthermore, the regulatory cell(s) appear to involve BoCD8+ T, lymphocytes which are specific for B. abortus antigens.
通过应用一种改良的检测方法来测量人类抑制性T淋巴细胞活性,并通过体外培养和鉴定抗原刺激的T淋巴细胞系,研究了牛对布鲁氏菌19号菌株(S19)免疫的免疫反应调节。总共9头公牛被分为两组:对照组(n = 4)和S19免疫组(n = 5)。在实验的第4天至第49天,定期将每只动物的外周血单核细胞(PBMC)与丝裂原(刀豆球蛋白A(Con A)和商陆有丝分裂原(PWM))、流产布鲁氏菌抗原(流产布鲁氏菌可溶性抗原(BASA)和全热灭活流产布鲁氏菌细胞(HKC))以及单独的培养基在体外培养。通过将这些培养物的上清液添加到用亚最佳浓度的Con A刺激的指示性培养物中,来检测其免疫调节活性。与对照组公牛在免疫后第35天和第49天的情况相比,用流产布鲁氏菌抗原培养的S19免疫公牛的PBMC产生的上清液显著(P小于0.05)抑制了指示细胞的反应。免疫牛的PBMC相对于对照牛的这种抑制活性也可以通过Con A或PWM的丝裂原刺激来诱导。在研究的第49天,免疫牛的PBMC自发释放出抑制活性。在免疫后2周和9周,从两头S19免疫的公牛中启动了T淋巴细胞系。通过体外检测对流产布鲁氏菌抗原的增殖反应以及通过使用一组有限的单克隆抗体进行间接免疫荧光来表征这些T细胞系的表面标志物表达。本研究结果表明,S19免疫诱导了牛PBMC中能够调节体外对流产布鲁氏菌反应的细胞亚群。这种调节活性在免疫后7周即可通过体外检测到。此外,调节细胞似乎涉及对流产布鲁氏菌抗原具有特异性的BoCD8 + T淋巴细胞。