Lawman M J, Griebel P, Hutchings D L, Davis W C, Heise J, Qualtiere L, Babiuk L A
Veterinary Infectious Disease Organization, Saskatoon, Saskatchewan, Canada.
Viral Immunol. 1987;1(3):163-76. doi: 10.1089/vim.1987.1.163.
To gain insight into the mechanisms of immunity to bovine herpesvirus type-1 (BHV-1) in particular the importance of the T-cell response, we attempted to clone bovine cytotoxic T lymphocytes that were specific for BHV-1 infected autologous target cells. A number of bovine T cell clones were generated by limiting dilution in the presence of bovine recombinant IL-2 and BHV-1 infected target cells as feeder layers. These clones were maintained in culture on crude IL-2 containing supernatants. In functional studies, 4 of the 16 T cell clones were shown to have high levels of cytotoxic activity specific for autolgous BHV-1 infected target cells with significantly lower cytotoxic activity against uninfected target cells and heterologous BHV-1 infected target cells. Continuous culturing of these 4 T cell clones, using either the crude IL-2 or high concentrations of recombinant bovine IL-2, resulted in the loss of both MHC restricted and BHV-1 specific cytotoxic activity. These clones now exhibit promiscuous type cytotoxic activity with the ability to lyse a variety of target cells. Using flow cytometric analysis, the phenotype of the T cell clones were shown to have bovine T lymphocyte characteristics including expression of the BoT8 marker. This is the first report of cloned bovine cytotoxic T lymphocytes reactive against BHV-1 and the generation from these clones of promiscuous cytotoxic activity against both virus-infected and non-infected bovine target cells.
为深入了解牛疱疹病毒1型（BHV - 1）的免疫机制，特别是T细胞反应的重要性，我们试图克隆对感染BHV - 1的自体靶细胞具有特异性的牛细胞毒性T淋巴细胞。通过在含有牛重组白细胞介素-2（IL - 2）和感染BHV - 1的靶细胞作为饲养层的情况下进行有限稀释，产生了许多牛T细胞克隆。这些克隆在含有粗制IL - 2的上清液中进行培养。在功能研究中，16个T细胞克隆中的4个显示出对自体感染BHV - 1的靶细胞具有高水平的细胞毒性活性，而对未感染的靶细胞和异源感染BHV - 1的靶细胞的细胞毒性活性明显较低。使用粗制IL - 2或高浓度的重组牛IL - 2对这4个T细胞克隆进行连续培养，导致MHC限制和BHV - 1特异性细胞毒性活性丧失。这些克隆现在表现出混杂型细胞毒性活性，能够裂解多种靶细胞。通过流式细胞术分析，T细胞克隆的表型显示具有牛T淋巴细胞特征，包括BoT8标志物的表达。这是关于克隆的对BHV - 1有反应的牛细胞毒性T淋巴细胞以及从这些克隆产生对病毒感染和未感染的牛靶细胞均具有混杂细胞毒性活性的首次报道。
