Gansuvd Balgansuren, Hubbard William J, Hutchings Anne, Thomas Francis T, Goodwin Jeanine, Wilson S Brian, Exley Mark A, Thomas Judith M
Department of Surgery, Division of Transplantation Immunobiology, University of Alabama, Birmingham, AL 35294, USA.
J Immunol. 2003 Sep 15;171(6):2904-11. doi: 10.4049/jimmunol.171.6.2904.
Natural killer T cells are immunoregulatory cells, which have important roles in tolerance and autoimmunity, as demonstrated primarily in mice and humans. In this study, we define the phenotype and function of Valpha24(+) T cells derived from the spleens of rhesus macaques, a species increasingly used in models of immune tolerance. Valpha24(+) cells were isolated and expanded with monocyte-derived immature dendritic cells in the presence of alpha-galactosylceramide, IL-2, and IL-15. Rhesus NKT cells were stained with mAbs against both Valpha24 and the invariant complementarity-determining region 3 epitope of the human Valpha24/JalphaQ TCR. The cells were CD4, CD8 double negative and expressed CD56. Rhesus NKT cells also exhibited moderate to high expression of CD95, CD45RO, CD11a, and beta(7) integrin, but did not express CD45 RA, CD62L, CCR7, CD28, and other activation, costimulatory molecules (CD69 and CD40L). By intracellular staining, >90% of unstimulated rhesus NKT cells expressed IL-10, but not IFN-gamma. However, the latter was strongly expressed after stimulation. Rhesus NKT secreted large amounts of TGF-beta, IL-13, and IL-6, and modest levels of IFN-gamma, whereas IL-10 secretion was negligible and no detectable IL-4 was observed either intracellularly or in culture supernatants. Functionally, the NKT cells and their supernatants suppressed T cell proliferation in allogeneic MLR. We conclude that long-term cultured rhesus macaque spleen-derived Valpha24(+) T cells are semi-invariant double-negative cells with effector memory phenotype. These cells are semianergic, polarized to a uniquely Th3 > T regulatory-1 regulatory cell phenotype, and have regulatory/suppressive function in vitro.
自然杀伤T细胞是免疫调节细胞，主要在小鼠和人类中得到证实，其在耐受性和自身免疫中发挥重要作用。在本研究中，我们定义了恒河猴脾脏来源的Vα24(+) T细胞的表型和功能，恒河猴这一物种越来越多地用于免疫耐受模型。Vα24(+)细胞在α-半乳糖神经酰胺、白细胞介素-2和白细胞介素-15存在的情况下，与单核细胞来源的未成熟树突状细胞一起分离和扩增。恒河猴NKT细胞用抗Vα24和人类Vα24/JαQ TCR的恒定互补决定区3表位的单克隆抗体进行染色。这些细胞为CD4、CD8双阴性，并表达CD56。恒河猴NKT细胞还表现出CD95、CD45RO、CD11a和β7整合素的中度至高度表达，但不表达CD45 RA、CD62L、CCR7、CD28以及其他激活、共刺激分子（CD69和CD40L）。通过细胞内染色，>90%未受刺激的恒河猴NKT细胞表达白细胞介素-10，但不表达干扰素-γ。然而，后者在刺激后强烈表达。恒河猴NKT分泌大量转化生长因子-β、白细胞介素-13和白细胞介素-6，以及适量的干扰素-γ，而白细胞介素-10的分泌可忽略不计，在细胞内或培养上清液中均未观察到可检测到的白细胞介素-4。在功能上，NKT细胞及其上清液在同种异体混合淋巴细胞反应中抑制T细胞增殖。我们得出结论，长期培养的恒河猴脾脏来源的Vα24(+) T细胞是具有效应记忆表型的半不变双阴性细胞。这些细胞处于半失能状态，极化为独特的Th3>T调节性1调节细胞表型，并在体外具有调节/抑制功能。
