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Heterogeneity and plasticity of porcine alveolar macrophage and pulmonary interstitial macrophage isolated from healthy pigs in vitro

机译：从健康猪中分离出的猪肺泡巨噬细胞和肺间质巨噬细胞的异质性和可塑性

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This study investigated the heterogeneity and plasticity of porcine alveolar macrophages (PAM) and pulmonary interstitial macrophages (IM) isolated from healthy pigs, including phenotype, function and gene expression. Dynamic changes of nitric oxide (NO) levels secreted by PAM and IM with stimulation of different doses of lipopolysaccharide (LPS) were investigated by Griess method, and the viability of the PAM and IM cells was investigated by MTT assay. Flow cytometry, fluorescence quantitative PCR and ELISA techniques were used to measure cell phenotype, gene expression and cytokine secretion, respectively. The PAM and IM cells in normal healthy pigs showed heterogeneity with 95.42±1.51% and 31.99±5.84% of CD163+ macrophage, respectively. The NO level in IM was significantly higher versus PAM after LPS treatment. Consistently, the ratio of Arg I/iNOS in IM was much lower than that in PAM, suggesting that the PAM belong to M2 macrophages and the IM belong to M1 macrophages. The PAM and IM cells in normal healthy pigs also showed plasticity. The Arg I/iNOS ratio and TIMP1/MMP12 ratio were significantly decreased in LPS- or LPS+IFNγ-treated PAM and IM, suggesting that cells were polarized towards M1 macrophages under LPS or LPS+IFNγ stimulation. On the contrary, IL-4 and IL-13 stimulation on PAM and IM lead to M2 polarization. A similar result was found in IL-1β gene expression and TNFα secretion. In conclusion, porcine macrophages have shown heterogeneity and plasticity on polarization under the stimulation of LPS, IFNγ, IL-4 and IL-13.
机译：本研究调查了从健康猪中分离出的猪肺泡巨噬细胞（PAM）和肺间质巨噬细胞（IM）的异质性和可塑性，包括表型，功能和基因表达。用Griess方法研究了不同剂量脂多糖（LPS）刺激下PAM和IM分泌的一氧化氮（NO）水平的动态变化，并通过MTT法研究了PAM和IM细胞的活力。流式细胞仪，荧光定量PCR和ELISA技术分别用于测量细胞表型，基因表达和细胞因子分泌。正常健康猪的PAM和IM细胞异质性分别为CD163 +巨噬细胞的95.42±1.51％和31.99±5.84％。 LPS治疗后，IM中的NO水平明显高于PAM。一致的是，IM中的Arg I / iNOS比率远低于PAM中的比率，这表明PAM属于M2巨噬细胞，而IM属于M1巨噬细胞。正常健康猪的PAM和IM细胞也显示可塑性。在LPS或LPS +IFNγ处理的PAM和IM中，Arg I / iNOS比值和TIMP1 / MMP12比值显着降低，表明在LPS或LPS +IFNγ刺激下，细胞向M1巨噬细胞极化。相反，PAM和IM上的IL-4和IL-13刺激导致M2极化。在IL-1β基因表达和TNFα分泌中发现了相似的结果。总之，在LPS，IFNγ，IL-4和IL-13的刺激下，猪巨噬细胞在极化上表现出异质性和可塑性。

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