Isolation and characterized of the porcine NKT cells

在人类和小鼠中，NKT细胞被表征为可以被糖脂抗原刺激的群体，如α-半乳糖神经酰胺（α-GalCer），其由CD 1d分子呈递。活化的NKT细胞可以产生细胞因子（IFN-γ和IL-4）和细胞溶解蛋白（穿孔素），表明该细胞群在先天性和获得性免疫中具有重要作用。本研究尝试对猪NKT细胞进行分离、培养和功能分析。流式细胞仪分析表明，在猪NK细胞（CD 8 ^+ CD 16 ^+）群体中存在表达T细胞标志物（CD 3 ^+）的细胞。此外，我们在猪CD 3 ^+细胞群中检测到小鼠CD 1d四聚体反应细胞。此外，我们还从猪PBMC中获得了人NKT细胞标志物TCRVα24和TCRVβ11的同源序列。获得的猪TcR序列中保守的氨基酸序列是NKT细胞与α-GalCer和CD 1d结合的重要氨基酸序列。这些结果有力地证明了猪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.