de Vries P, Versteeg-van Oosten J P, Visser I K, van Binnendijk R S, Langeveld S A, Osterhaus A D, Uytdehaag F G
Department of Immunobiology, National Institute of Public Health and Environmental Protection, Bilthoven, The Netherlands.
J Immunol. 1989 Apr 15;142(8):2841-6.
Measles virus (MV)-specific murine helper T cell clones (Thy-1.2+, CD4+, CD8-) were generated from mice immunized with MV-infected mouse brain homogenate by limiting dilution and in vitro stimulation of spleen cells with UV-inactivated MV Ag. The protein specificity of 7 out of 37 stable T cell clones, which displayed MHC-restricted MV Ag recognition, could be assessed by using purified MV proteins. Two fusion (F) protein-specific, two hemagglutinin-specific, and three nucleoprotein- or matrix protein-specific clones were shown to be established. The F protein-specific T cell clones together with a panel of previously generated F protein-specific T cell clones were characterized for their fine specificity by using beta-galactosidase fusion products, which contained different parts of the F protein. It was shown that at least two epitopes on the major part of the F protein (amino acid 2-513) can be recognized by mouse T cells. Functional characterization of three T cell clones showed that they were able to assist MV-specific B cells and bystander B cells for antibody production. Furthermore, they were shown to produce the lymphokines IL-2 and IFN-gamma. It was also shown that these T cell clones induced a MV-specific delayed type hypersensitivity response. These observations suggest that all of the T cell clones characterized belong to the TH1 helper subset.
通过有限稀释法,并用紫外线灭活的麻疹病毒(MV)抗原体外刺激脾细胞,从用MV感染的小鼠脑匀浆免疫的小鼠中产生了麻疹病毒(MV)特异性小鼠辅助性T细胞克隆(Thy-1.2 +、CD4 +、CD8 -)。37个稳定的T细胞克隆中有7个表现出MHC限制的MV抗原识别,其蛋白质特异性可通过使用纯化的MV蛋白来评估。已证明建立了两个融合(F)蛋白特异性、两个血凝素特异性以及三个核蛋白或基质蛋白特异性的克隆。通过使用含有F蛋白不同部分的β-半乳糖苷酶融合产物,对F蛋白特异性T细胞克隆以及一组先前产生的F蛋白特异性T细胞克隆进行了精细特异性表征。结果表明,小鼠T细胞可识别F蛋白主要部分(氨基酸2 - 513)上至少两个表位。三个T细胞克隆的功能表征表明,它们能够辅助MV特异性B细胞和旁观者B细胞产生抗体。此外,它们还被证明可产生细胞因子白细胞介素-2(IL-2)和干扰素-γ(IFN-γ)。还表明这些T细胞克隆可诱导MV特异性迟发型超敏反应。这些观察结果表明,所表征的所有T细胞克隆均属于TH1辅助亚群。