Role of the Cytokine Profiles Produced by iNKT Cells in the Initial Phase of Cyclophosphamide-Induced Tolerance^1

环磷酰胺（CP）诱导的耐受是一种基于混合嵌合体的耐受诱导方案。最近，我们报道了恒定的自然杀伤T（iNKT）细胞在该系统中的耐受诱导是必不可少的。在这项研究中，我们评估了iNKT细胞产生的细胞因子的作用。DBA/2（H-2_d）小鼠和BALB/c（H-2_d）野生型（WT）或iNKT敲除（KO）小鼠用作供体和受体。WT接受者接受三次剂量（第-7天）。-4、-1或35、38、41）或单剂量（第-1或0天）的α-半乳糖神经酰胺（GC）与我们的预处理方案联合，预处理方案由第0天10^8个供体脾细胞（SC）和第2天200 mg/kg CP组成。为了研究iNKT细胞功能，将iNKT KO受体用细胞因子（IFN-γ）重建。IL-4或IL-10）KO iNKT细胞并接受供体SC和CP。在WT受体中观察到混合嵌合体，但在iNKT KO受体中减少。然而，在第-7、-4、-1天给予GC的WT受体中不存在混合嵌合体，但在第35、38、41天给予GC的WT受体中不存在。当混合嵌合体减少时，供体皮肤移植物被慢性排斥。在用来自IFN-γ、IL-4或IL-10 KO小鼠的iNKT细胞重建并接受我们的预处理方案的iNKT KO受体中接受皮肤移植物。在嵌合体诱导的初始阶段需要iNKT细胞。我们的研究结果表明，已知的主要细胞因子产生的iNKT细胞的调节功能的iNKT细胞。

Cyclophosphamide (CP) -induced tolerance is a mixed chimerism-based tolerance induction protocol. Recently, we reported that invariant natural killer T (iNKT) cells were essential for the tolerance induction in this system. In this study we evaluated the roles of the cytokines produced by iNKT cells. DBA/2 (H-2^d) mice and BALB/c(H-2^d)wild-type (WT) or iNKT knockout (KO) mice were used as donors and recipients. WT recipients received three doses(days -7. -4, -1 or 35, 38, 41)or a single dose (day -1 or 0) of a-galactosylceramide (GC) in conjunction with our conditioning regimen, which consisted of 10^8 donor spleen cells (SC) on day 0 and 200 mg/kg CP on day 2. To investigate the iNKT cell function, iNKT KO recipients were reconstituted with cytokine(IFN-y. IL-4,or IL-10)KO iNKT cells and received donor SC and CP.Mixed chimerism was observed in WT recipients, but was reduced in iNKT KO recipients. However, mixed chimerism was absent in WT recipients given GC on days -7, -4, -1, but not in WT recipients given GC on day 35, 38, 41. Donor skin grafts were chronically rejected when mixed chimerism was diminished. Skin grafts were accepted in iNKT KO recipients reconstituted with iNKT cells from IFN-y, IL-4, or IL-10 KO mice and receiving our conditioning regimen. iNKT cells were required in the initial phase of the induction of chimerism. Our results indicated that known major cytokines produced by iNKT cells were dispensable for the regulatory function of iNKT cells.