Nataraj C, Srikumaran S
Department of Veterinary and Biomedical Sciences, IANR, University of Nebraska-Lincoln.
Viral Immunol. 1994;7(1):11-23. doi: 10.1089/vim.1994.7.11.
Difficulties in the isolation and long-term maintenance of bovine herpesvirus-1 (BHV-1) specific T-cell clones have hindered the analysis of bovine cell-mediated immune response to this virus. In an effort to identify the T-cell epitopes of the virus, bovine murine T-cell hybridomas specific for BHV-1 were generated as an alternative to T-cell clones. Peripheral blood lymphocytes from a calf immunized with BHV-1 were restimulated in vitro with the virus to generate bulk T-cell cultures. The antigen-specific T-cell-enriched bulk culture lymphocytes were fused with the T-cell receptor-deficient mutant of the murine thymoma cell line BW 5147. T-cell hybridomas were screened for their ability to produce interferon-gamma in response to BHV-1 stimulation. Hybridomas with various specificities were obtained. One of them was specific for the BHV-1 glycoprotein gI, two were specific for gIV, while three other hybridomas were specific for gIII. One hybridoma responded to stimulation with BHV-1, but not to any of the glycoproteins gI, gIII, or gIV, suggesting that proteins other than these major glycoproteins may be involved in the bovine T-cell response to BHV-1. Of these hybridomas, one was MHC Class I restricted, while all the others were Class II restricted.
牛疱疹病毒1型（BHV-1）特异性T细胞克隆的分离及长期维持存在困难，这阻碍了对牛针对该病毒的细胞介导免疫反应的分析。为了鉴定该病毒的T细胞表位，制备了对BHV-1具有特异性的牛鼠T细胞杂交瘤，作为T细胞克隆的替代方法。用BHV-1免疫的小牛的外周血淋巴细胞在体外经该病毒再次刺激，以产生大量T细胞培养物。将富含抗原特异性T细胞的大量培养淋巴细胞与鼠胸腺瘤细胞系BW 5147的T细胞受体缺陷型突变体融合。筛选T细胞杂交瘤对BHV-1刺激产生γ干扰素的能力。获得了具有不同特异性的杂交瘤。其中一个对BHV-1糖蛋白gI具有特异性，两个对gIV具有特异性，而另外三个杂交瘤对gIII具有特异性。一个杂交瘤对BHV-1刺激有反应，但对糖蛋白gI、gIII或gIV中的任何一种都无反应，这表明除这些主要糖蛋白外的其他蛋白质可能参与牛对BHV-1的T细胞反应。在这些杂交瘤中，一个受MHC I类限制，而其他所有杂交瘤都受II类限制。
