Establishment of antigen specific regulatory dendritic cells and treatment of VKH disease with these dendritic cells

我们分析了体外预冲的树突状细胞是否具有诱导自身免疫性疾病的能力。用抗CD 11 c磁珠法收集C57 BL/6小鼠树突状细胞，在含GM-CSF的RPMI 1640中培养。用TRP-1肽脉冲这些细胞。将这些TRP-1肽预致敏的树突状细胞与CD 40和LPS进一步培养。将预致敏和成熟的树突状细胞注射到未处理的C57 BL/6小鼠中，注射这些树突状细胞的小鼠发展为自身免疫性疾病。临床表现为皮肤色素脱失、白发。组织学上，毛囊周围及部分腺体周围可见大量炎性细胞浸润。皮损处可见皮肤溃疡，炎性细胞浸润至真皮及真皮下。在色素脱失和/或白发区，肉芽肿由含有大量色素的大细胞形成，在眼内组织中，肉芽肿形成于虹膜和脉络膜。这些结果与VKH病的表现非常相似。接下来，我们用IL-10和TGF-β培养预冲的树突状细胞，以保持未成熟阶段。将这些未成熟的细胞注射到幼稚小鼠中，并测试细胞的抑制作用。与我们的推测相反，通过注射这些未成熟的树突状细胞没有检测到抑制作用。我们正在研究这些结果的机制。

We have analyzed whether in vitro prepulsed dendritic cells have the capability to induce the autoimmune disease. The dendritic cells of C57BL/6 mice were collected with anti-CD11c magnetic beads system and cultured in RPMI1640 containing GM-CSF. These cells were pulsed with TRP-1 peptides. These TRP-1 peptides prepulesd dendritic cells were further cultured with CD40 and LPS. Thus prepulsed and matured dendritic cells were injected into naive C57BL/6 mice.The mice injected with these dendritic cells had developed autoimmune disease. The clinical findings of this disease were featured with depigmentation and poliosis of skin. However inflammatory changes of the intraocular tissues were not clearly seen by ordinally observation.Histologically, many inflammatory cells were seen around the hair follicles and some glands. Skin ulcers were seen in the lesions where the inflammatory cells infiltrate into the dermis and under the dermis. In the depigmented and/or poliosis regions, granulomas were formed with larges cells that contain many pigments.In the intra ocular tissues, granulomas were formed in the iris and choroid. All these findings were very similar to the findings of VKH disease.Next, we cultured the prepulsed dendritic cells with IL-10 and TGF-β to keep immature stage. These immature cells were injected to naive mice and the inhibitory effects of the cells were tested. In contrast to our speculation, no inhibitory effects were detected by injection of these immature dendritic cells. We are now studying the mechanism of these results.