Department of Critical Care Units, The First Affiliated Hospital, Zhejiang University, School of Medicine, Hangzhou, China.
State Key Laboratory for Diagnosis & Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University, School of Medicine, Hangzhou, China.
Front Cell Infect Microbiol. 2021 Jan 18;10:606340. doi: 10.3389/fcimb.2020.606340. eCollection 2020.
The macrophage stimulating protein (MSP)-Recepteur d'origine nantais (RON) signaling pathway regulates macrophage function. Here, we verified RON receptor expression in bone marrow-derived dendritic cells (BMDCs) by real time-PCR, Western blot, and flow cytometry. Flow cytometry was used to detect the changes in MHC II and CD86 expression following the inhibition of RON in BMDCs and splenic dendritic cells (DCs). Immunoprecipitation and Western blot were used to detect the level of MHC II and CD86 ubiquitination. An enzyme-linked immunosorbent assay was used to detect cytokine release, and a mixed lymphocyte reaction was performed to evaluate DC maturity. The results show that the inhibition of RON leads to an increase in March-1 transcription, which intensifies the ubiquitination of MHC II and CD86 and ultimately leads to a decreased level of these two molecules. The mixed lymphocyte reaction provided evidence that RON inhibition decreased the ability of DCs to promote the proliferation of T cells. The MSP-RON signaling pathway may play an important role in lipopolysaccharide (LPS)-stimulated DC maturation through March-I and may protect DC differentiation following LPS stimulation.
巨噬细胞刺激蛋白 (MSP)-纳坦受体 (RON) 信号通路调节巨噬细胞功能。在此，我们通过实时 PCR、Western blot 和流式细胞术验证了 RON 受体在骨髓来源树突状细胞 (BMDC) 中的表达。流式细胞术用于检测 BMDC 和脾树突状细胞 (DC) 中 RON 抑制后 MHC II 和 CD86 表达的变化。免疫沉淀和 Western blot 用于检测 MHC II 和 CD86 泛素化水平。酶联免疫吸附试验用于检测细胞因子释放，混合淋巴细胞反应用于评估 DC 成熟度。结果表明，RON 的抑制导致 March-1 转录增加，从而增强 MHC II 和 CD86 的泛素化，最终导致这两种分子水平降低。混合淋巴细胞反应提供的证据表明，RON 抑制降低了 DC 促进 T 细胞增殖的能力。MSP-RON 信号通路可能通过 March-I 在脂多糖 (LPS) 刺激的 DC 成熟中发挥重要作用，并可能在 LPS 刺激后保护 DC 分化。
