关键词: 猪T淋巴细胞; 增殖反应; 流式细胞术; 刀豆蛋白A; MTT法

Abstract: Recognition of MHC-peptide complexes by T lymphocytes results in robust T cell proliferation, which is regarded as one of the hallmarks of the host cell-mediated immune responses against pathogens. To conveniently detect cell-mediated immune responses of swine upon pathogen infection or vaccine immunization, we developed a method for studying peripheral blood T cell proliferation of swine via MTT assay. Firstly, peripheral blood mononuclear cells (PBMC) were collected through density gradient centrifugation, and then peripheral blood lymphocytes (PBL) were obtained by discarding adherent monocytes after co-culture overnight. Flow cytometry analysis showed that the percentage of T cells in PBL was >80%. Next, the MTT assay was used to detect effects of the concentration of ConA and cell culture density on porcine T cell proliferation. The result of MTT suggested that when 2×106/mL T cells were cultured with 5 μg/mL ConA, the highest level of T cell proliferation was achieved. The MTT assay for studying porcine T cell proliferation could facilitate assessing the T cell mediated immune response against pathogens or vaccines.

Key words: porcine T cells; proliferation; flow cytometry; ConA; MTT assay