Singh Inderpal, McConnell Ian, Blacklaws Barbara
Centre for Veterinary Science, Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge, United Kingdom CB3 0ES.
J Virol. 2006 Jan;80(2):912-9. doi: 10.1128/JVI.80.2.912-919.2006.
The lesions caused by maedi-visna virus (MVV) are known to be immune mediated with a presumed contribution by the response to viral antigens. However, very little is known about the T-cell response to individual viral proteins. We have therefore expressed the three individual gag antigens of MVV strain EV1 (p16, p25, and p14) in a bacterial expression system and used the purified recombinant proteins to analyze the antibody and CD4+ T-cell response to MVV. Plasma samples were taken from sheep after 1 year of infection with MVV. The titers for antibodies in these samples were determined by indirect enzyme-linked immunosorbent assays and were as follows: anti-p25 antibody, 1:400 to >1:3,200; anti-p16 antibody, 1:400 to 1:3,200; and anti-p14 antibody, 1:<100 to 1:3,200. When the induction of antibodies was followed over time postinfection (p.i.), samples positive for anti-p25 were seen by day 24 p.i., followed by anti-p16 by day 45 p.i., and lastly anti-p14 by day 100 p.i. T-cell proliferative responses to all three gag antigens were detected in persistently infected sheep peripheral blood lymphocytes. The antigens were therefore used to raise T-cell lines from persistently infected sheep. These T-cell lines were shown to be specific for the recombinant gag antigens and for viral antigen expressed on infected macrophages. The proliferative response was restricted to major histocompatibility complex class II HLA-DR and so was due to CD4+ T lymphocytes. All three gag antigens may therefore play a role in immune-mediated lesion formation in MVV disease by presentation on infected macrophages in lesions.
已知梅迪-维斯纳病毒(MVV)引起的病变是免疫介导的,推测病毒抗原的反应起了作用。然而,关于T细胞对单个病毒蛋白的反应知之甚少。因此,我们在细菌表达系统中表达了MVV毒株EV1的三种单个gag抗原(p16、p25和p14),并使用纯化的重组蛋白分析对MVV的抗体和CD4+ T细胞反应。在绵羊感染MVV 1年后采集血浆样本。通过间接酶联免疫吸附测定法测定这些样本中的抗体滴度,结果如下:抗p25抗体,1:400至>1:3200;抗p16抗体,1:400至1:3200;抗p14抗体,1:<100至1:3200。当在感染后(p.i.)随时间追踪抗体诱导情况时,在感染后第24天观察到抗p25呈阳性的样本,随后在感染后第45天观察到抗p16呈阳性的样本,最后在感染后第100天观察到抗p14呈阳性的样本。在持续感染的绵羊外周血淋巴细胞中检测到对所有三种gag抗原的T细胞增殖反应。因此,使用这些抗原从持续感染的绵羊中培养T细胞系。这些T细胞系显示对重组gag抗原和感染巨噬细胞上表达的病毒抗原具有特异性。增殖反应限于主要组织相容性复合体II类HLA-DR,因此是由CD4+ T淋巴细胞引起的。因此,所有三种gag抗原可能通过在病变中的感染巨噬细胞上呈递,在MVV疾病的免疫介导病变形成中发挥作用。