Flynn J N, Cannon C A, Beatty J A, Mackett M, Rigby M A, Neil J C, Jarrett C
Department of Veterinary Pathology, University of Glasgow, Bearsden, United Kingdom.
J Virol. 1994 Sep;68(9):5835-44. doi: 10.1128/JVI.68.9.5835-5844.1994.
The role of cellular immunity in the establishment and progression of immunosuppressive lentivirus infection remains equivocal. To develop a model system with which these aspects of the host immune response can be studied experimentally, we examined the response of cats to a hybrid peptide containing predicted T-and B-cell epitopes from the gag and env genes of feline immunodeficiency virus (FIV). Cats were immunized with an unmodified 17-residue peptide incorporating residues 196 to 208 (from gag capsid protein p24) and 395 to 398 (from env glycoprotein gp120) of the FIV Glasgow-8 strain by using Quil A as an adjuvant. Virus-specific lymphocytotoxicity was measured by chromium-51 release assays. The target cells were autologous or allogeneic skin fibroblasts either infected with recombinant FIV gag vaccinia virus or pulsed with FIV peptides. Effector cells were either fresh peripheral blood mononuclear cells or T-cell lines stimulated with FIV peptides in vitro. Cytotoxic effector cells from immunized cats lysed autologous, but not allogeneic, target cells when they were either infected with recombinant FIV gag vaccinia virus or pulsed with synthetic peptides comprising residues 196 to 205 or 200 to 208 plus 395. Depletion of CD8+ T cells, from the effector cell population abrogated the lymphocytotoxicity. Immunized cats developed an antibody response to the 17-residue peptide immunogen and to recombinant p24. However, no antibodies which recognized smaller constituent peptides could be detected. This response correlated with peptide-induced T-cell proliferation in vitro. This study demonstrates that cytotoxic T lymphocytes specific for FIV can be induced following immunization with an unmodified short synthetic peptide and defines a system in which the protective or pathological role of such responses can be examined.
细胞免疫在免疫抑制性慢病毒感染的发生和发展过程中的作用仍不明确。为了建立一个能够通过实验研究宿主免疫反应这些方面的模型系统，我们检测了猫对一种混合肽的反应，该混合肽包含来自猫免疫缺陷病毒（FIV）gag和env基因的预测T细胞和B细胞表位。用含有FIV格拉斯哥-8株gag衣壳蛋白p24的196至208位残基和env糖蛋白gp120的395至398位残基的未修饰17残基肽免疫猫，使用Quil A作为佐剂。通过51铬释放试验测量病毒特异性淋巴细胞毒性。靶细胞是感染重组FIV gag痘苗病毒或用FIV肽脉冲处理的自体或同种异体皮肤成纤维细胞。效应细胞是新鲜外周血单核细胞或体外经FIV肽刺激的T细胞系。当免疫猫的细胞毒性效应细胞感染重组FIV gag痘苗病毒或用包含196至205位残基或200至208位残基加395位残基的合成肽脉冲处理时，可裂解自体靶细胞，但不能裂解同种异体靶细胞。从效应细胞群体中去除CD8 + T细胞可消除淋巴细胞毒性。免疫猫对17残基肽免疫原和重组p24产生了抗体反应。然而，未检测到识别较小组成肽的抗体。这种反应与体外肽诱导的T细胞增殖相关。本研究表明，用未修饰的短合成肽免疫后可诱导出针对FIV的细胞毒性T淋巴细胞，并定义了一个可研究此类反应的保护或病理作用的系统。
