Isolation and characterized of the porcine NKT cells

在人和小鼠中，NKT细胞是一个能被糖脂抗原刺激的群体，如CD1d分子提出的α-半乳糖神经酰胺(α-GalCer)。活化的NKT细胞既能产生细胞因子(干扰素-γ和IL-4)，又能产生细胞溶解蛋白(穿孔素)，提示该细胞群在先天免疫和获得性免疫中均有重要作用。本研究试图对猪NKT细胞进行分离、培养和功能分析。流式细胞仪分析表明，猪NK细胞(CD8、CD16)群中存在T细胞标志物(CD3)表达细胞。此外，我们还在猪CD3细胞群中检测到小鼠CD1d四聚体反应性细胞。此外，我们还从猪外周血单核细胞中获得了人NKT细胞标志物TCRVα24和TCRVβ11的同源序列。与α-GalCer和CD1d结合的氨基酸序列在获得的猪TcR序列中是保守的。这些结果有力地提示了猪NKT细胞的存在。

In human and mouse, NKT cells are characterized as a population that can be stimulated by glycolipid antigen, such as α-Galactosylceramide (α-GalCer), which is presented by CD1d molecule. Activated NKT cells can produce both cytokines (IFN-γ and IL-4) and cytolytic protein (perforin), suggesting that this cell population have an important role in both innate and acquired immunity. In this study, we tried to conduct isolation, culture and functional analysis of porcine NKT cells. Flow cytometry analysis showed that there are T cell marker (CD3^+)-expressing cells in porcine NK cell (CD8^+CD16^+) population. In addition, we detected mouse CD1d tetramer-reactive cells in porcine CD3^+ cell population. Furthermore, we obtained the homologous sequence of human NKT cell markers, TCRVα24 and TCRVβ11 from pig PBMC. The amino acid sequences that are important for NKT cell binding to α-GalCer and CD1d were conserved in the obtained porcine TcR sequences. These results strongly suggested the existence of porcine NKT cells.