Dillender M J, Lunney J K
Helminthic Diseases Laboratory, Agricultural Research Service, USDA, Beltsville, MD 20705.
Vet Immunol Immunopathol. 1993 Jan;35(3-4):301-19. doi: 10.1016/0165-2427(93)90041-2.
MHC restricted, antigen specific, cytokine dependent T-cell lines were established from peripheral blood mononuclear cells (PBMC) of NIH minipigs that were challenge inoculated with Trichinella spiralis. Swine lymphocyte antigen (SLA) inbred NIH minipigs of the SLA a/a (aa) haplotype can be divided into Responder (R) and Non-responder (NR) phenotypes on the basis of their ability to destroy encysted muscle larval (ML) forms of the parasite T. spiralis. When orally inoculated with 300 infectious ML and challenge inoculated with 10,000 ML, R pigs are able to mount an immune attack against encysted ML resident in host muscle cells since the primary inoculation. Because T-lymphocyte mediated mechanisms are likely to be operative in this response, a series of T-lymphocyte cell lines was established from PBMC of ten aa minipigs 10 days after the challenge infection with T. spiralis. These T-lymphocyte lines showed antigen specific, cytokine dependent proliferation in response to T. spiralis antigen stimulation. T-lymphocyte cell lines from R and NR pigs were compared and found to be similar in the magnitude of proliferative responses, in the duration of antigen specificity, and in the alterations in T-lymphocyte subsets and cell surface phenotype after antigen stimulation. In vitro antigen stimulation of T-lymphocyte cell lines and evaluation of small lymphocytes and lymphoblasts by density gradient centrifugation, immunofluorescent staining and flow cytometry, revealed that up to 55% of the cells were lymphoblasts at 4 days after stimulation, and that the majority of the lymphoblast population expressed CD4 and CD8, as well as the interleukin-2 receptor and the Class II alloantigen, SLA-DR (DR+). These results indicate that pig T-lymphocytes of the CD4+CD8+ phenotype may be involved in the anamnestic cell mediated immune response to T. spiralis.
从经旋毛虫攻击接种的NIH小型猪外周血单个核细胞(PBMC)中建立了MHC限制、抗原特异性、细胞因子依赖性T细胞系。SLA a/a(aa)单倍型的NIH小型猪在猪淋巴细胞抗原(SLA)近交系中,可根据其破坏旋毛虫包囊化肌幼虫(ML)形式的能力分为反应者(R)和无反应者(NR)表型。当口服接种300条感染性ML并攻击接种10000条ML时,R猪自初次接种后就能对宿主肌肉细胞中驻留的包囊化ML发动免疫攻击。由于T淋巴细胞介导的机制可能参与此反应,在经旋毛虫攻击感染10天后,从10只aa小型猪的PBMC中建立了一系列T淋巴细胞系。这些T淋巴细胞系在受到旋毛虫抗原刺激后表现出抗原特异性、细胞因子依赖性增殖。对来自R猪和NR猪的T淋巴细胞系进行比较,发现它们在增殖反应强度、抗原特异性持续时间以及抗原刺激后T淋巴细胞亚群和细胞表面表型变化方面相似。通过密度梯度离心、免疫荧光染色和流式细胞术对T淋巴细胞系进行体外抗原刺激以及对小淋巴细胞和成淋巴细胞进行评估,结果显示刺激后4天高达55%的细胞为成淋巴细胞,并且大多数成淋巴细胞群体表达CD4和CD8,以及白细胞介素-2受体和II类同种异体抗原SLA-DR(DR+)。这些结果表明,CD4+CD8+表型的猪T淋巴细胞可能参与对旋毛虫的回忆性细胞介导免疫反应。