Cellular Analysis of Cutaneous Granuloma induced by Antigen-binding Agarose Particle and MDP

我们检测了抗原结合琼脂糖粒和MDP诱导的小鼠皮肤肉芽肿中的免疫细胞和细胞因子的产生。肉芽肿中央及周围有少量CD3+细胞。CD_4~+和CD_8~+细胞分布几乎相同。CD14+单核细胞是肉芽肿的主要细胞，但肉芽肿中央未见MMGL阳性的巨噬细胞。干扰素-γ+细胞明显多于IL-4+细胞，提示诱导的皮肤肉芽肿具有TH1型反应的特点。为了解GFP转基因小鼠外周血单核巨噬细胞系前体细胞在肉芽肿中的分布情况，制备了外周血单核细胞、单核细胞培养诱导的巨噬细胞、正常皮肤MMGL+真皮巨噬细胞、腹膜巨噬细胞、骨髓细胞、GM-CSF/IL-4诱导的骨髓细胞树突状细胞。这些细胞被皮下或静脉注射到带肉芽肿的小鼠体内。仅在注射新鲜分离的骨髓细胞时，肉芽肿中才发现GFP+细胞。为探讨肉芽肿上皮样细胞对细胞因子的反应，通过清除肉芽肿中CD3+细胞、B220+细胞和MMGL+细胞来丰富上皮样细胞。这些细胞对干扰素-γ和IL-3有反应，但对IL-13或IL-10无反应。这些结果提示骨髓中的浸润性细胞由抗原结合性琼脂糖粒和亚甲基二磷酸果糖诱导的皮肤肉芽肿组成，干扰素-γ和EL-3是肉芽肿形成的关键因素。

We examined immune cells and cytokine production in murine cutaneous granulomas induced by antigen-binding agarose particles and MDP. A few number of CD3+ cells are present in the center of granulomas and their surrounding areas. Almost same numbers of CD4+ and CD8+ cells are distributed. On the other hand, CD14+ monocytes are main cells of the granuloma, but tissure macrophages which are positive for MMGL were not present in the center of the granulomas. IFN-γ+ cells were more predominantly distributed than IL-4+ cells, suggesting that the induced cutaneous granuloma had a property of TH1 type reaction. To examine the propety of precursor cells of monocyte-macrophage lineage cells in the granulomas, the folowing cells were prepared from GFP-transgeneic mice, 1) peripheral monocytes, 2) macrophages induced by culturing monoctes, 3) MMGL+ dermal macrophages in normal skin, 4) peritoneal macrophages, 5) bone marrow cells, 6) dendritic cells induced from bone marrow cells with GM-CSF/IL-4. These cells were administered subcutaneously or intraveneously into granuloma-bearing mice. GFP+ cells were found in the granuloma only when freshly isolated bone marrow cells were injected. To investigate the response ofepithelioid cells in granuloma to cytokines, epithelioid cells were riched by eliminating CD3+ cells, B220+ cells and MMGL+ cells in the granulomas. Theses cells responded to IFN-γ and IL-3 but not to IL-13 or IL-10. These results suggest that infiltrating cells from bonw marrows consist of cutaneous granulomas induced by antigen-binding agarose particles and MDP, and that IFN-γ and EL-3 are a key factor for the formation of granulomas.