Weynants V, Walravens K, Didembourg C, Flanagan P, Godfroid J, Letesson J J
Laboratoire d'Immunologie-Microbiologie, U.R. Biologie Moléculaire, Facultés Universitaires Notre-Dame de la Paix, Namur, Belgium.
Vet Immunol Immunopathol. 1998 Dec 11;66(3-4):309-20. doi: 10.1016/s0165-2427(98)00205-0.
Peripheral blood mononuclear cells (PBMC) isolated from cattle infected with Brucella secreted gamma-interferon (IFN-gamma) after antigen-specific stimulation with Brucellergene, which is a mixture of cytoplasmic proteins of rough Brucella melitensis B115. Following the depletion of the monocyte-macrophages from the PBMC, the enriched lymphocyte populations stimulated with Brucellergene did not produce IFN-gamma. Two-colour immunofluorescence staining of intracellular IFN-gamma and bovine cell surface molecules identified the cells producing IFN-gamma among the PBMC stimulated with Brucellergene. Moreover, this method could be used to estimate the number of T-cells specifically producing IFN-gamma. For a given animal, there is a significant correlation (p < 0.05) between the production of IFN measured by an ELISA of the supernatant of whole blood stimulated with Brucellergene and the number of T-cells producing IFN-gamma after in vitro stimulation with Brucellergene. The development of the immunofluorescence staining technique provides a new tool for analysing and for measuring the T-cell immune response in cattle.
从感染布鲁氏菌的牛中分离出的外周血单个核细胞（PBMC），在用粗糙型布鲁氏菌羊种B115的胞质蛋白混合物布鲁氏菌素进行抗原特异性刺激后，分泌γ-干扰素（IFN-γ）。从PBMC中去除单核细胞-巨噬细胞后，用布鲁氏菌素刺激富集的淋巴细胞群体不产生IFN-γ。细胞内IFN-γ和牛细胞表面分子的双色免疫荧光染色确定了在用布鲁氏菌素刺激的PBMC中产生IFN-γ的细胞。此外，该方法可用于估计特异性产生IFN-γ的T细胞数量。对于给定的动物，用布鲁氏菌素刺激全血上清液的ELISA法测得的IFN产量与用布鲁氏菌素体外刺激后产生IFN-γ的T细胞数量之间存在显著相关性（p < 0.05）。免疫荧光染色技术的发展为分析和测量牛的T细胞免疫反应提供了一种新工具。
