LysM Cre Cbl-b/c-Cbl小鼠模型的建立和免疫表型分析-东南大学学报医学版

LysM Cre Cbl-b/c-Cbl小鼠模型的建立和免疫表型分析

单位：苏州大学生物医学研究院, 江苏苏州 215123

分类号：R392.1

出版年·卷·期（页码）：2017·36·第六期（978-984）

摘要：

目的：为深入研究Cbl-b/c-Cbl在巨噬细胞分化发育中的作用，拟建立LysM Cre Cbl-b/c-Cbl小鼠模型，并初步分析其免疫表型。方法：将LysMCre小鼠和Cbl-b^-/-及c-Cbl^f/f小鼠杂交得到LysM Cre⁺ Cbl-b^-/-/c-Cbl^f/f小鼠，从而在巨噬细胞中同时敲除Cbl-b和c-Cbl。并通过PCR和Western-blot鉴定模型建立的成功，通过HE、Giemsa及天狼星红染色和流式细胞仪分析LysM Cre⁺ Cbl-b^-/-/c-Cbl^f/f小鼠免疫表型。结果：通过PCR和Western-blot鉴定LysM Cre⁺ Cbl-b^-/-/c-Cbl^f/f模型建立成功，表型分析提示Cbl-b和c-Cbl在髓样来源细胞敲除后，导致DC、粒细胞和巨噬细胞比例增多，尤其是肺泡巨噬细胞在肺泡中大量侵润，其绝对数是WT小鼠的几百倍，且最终LysM Cre⁺ Cbl-b^-/-/c-Cbl^f/f小鼠因肺纤维化而死亡。结论：Cbl-b和c-Cbl在巨噬细胞的分化发育中发挥极其重要的作用，其同时在巨噬细胞中缺失，可以导致肺巨噬细胞的急剧增多，引起炎症和肺纤维化而导致小鼠死亡。

Objective: To explore the function of Cbl-b/c-Cbl in the development and differentiation of macrophage, and to establish the mouse model called LysM Cre⁺ Cbl-b^-/-/c-Cbl^f/f which is ablation both of Cbl-b and c-Cbl. Methods: The mice of LysMCre⁺, Cbl-b^-/- and c-Cbl^f/f were breed together, then PCR and Western-blot were conducted to identify the success of establishment of mouse model. HE, Giemsa and Sirus Red staining and FACS were applied to analyze the immune phenotype of these mice. Results: We successfully established the LysMCre⁺, Cbl-b^-/- and c-Cbl^f/f mouse model confirmed by PCR and Werstern-blot. Ablation of both Cbl-b and c-Cbl in myloid-derived cells resulted in the percentage increase of cDC,granucyte and macrophage. Obviously, a lot of alveolar macrophages were immersed in the lung, thus induced severe inflammation, finally developed into idiopathic pulmonary fibrosis, and mice died 35 day about after birth. Conclusions: Cbl-b and c-Cbl play an important role in macrophage, ablation both of Cbl-b and c-Cbl results in idiopathic pulmonary fibrosis of mice.

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