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. 2019 Oct 24;8(10):bio046342.
doi: 10.1242/bio.046342.

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

Huan Liu  1 , Jia Liu  2 , Jing Huang  1 , Xianchang Bai  1 , Qinfu Wang  3
Affiliations

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

Huan Liu et al. Biol Open. .

Abstract

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.

Keywords: Heterogeneity; Plasticity; Porcine alveolar macrophages; Pulmonary interstitial macrophages.

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Conflict of interest statement

Competing interestsThe authors declare no competing or financial interests.

Figures

Fig. 1.
Fig. 1.
CD163+ rates on PAM cells and IM cells isolated from healthy pigs. Gray area represents the surface CD163 expression levels in PAM cells (A) and IM cells (B) and the white area represents the isotype control. **P<0.01.
Fig. 2.
Fig. 2.
Dynamic changes of NO levels in PAM and IM cells stimulated by different doses of LPS (A) and the viability of the PAM and IM cells (B) in vitro. *P<0.05, **P<0.01.
Fig. 3.
Fig. 3.
Arg I/iNOS ratios in PAM and IM cells treated with LPS versus control.
Fig. 4.
Fig. 4.
Plasticity of PAM cells and IM cells isolated from healthy pigs after initial and secondary treatment in vitro. Plasticity of PAM cells and IM cells isolated from healthy pigs was described by the IL-1β/HPRT relative expression (A,B), the Arg1/iNOS ratio (C,D), TIMP/MMP12 ratio (E,F). PAM cells were shown in the left panel (A,C,E) and IM cells were shown in the right panel (B,D,F). *P<0.05, **P<0.01.
Fig. 5.
Fig. 5.
TNFα production of PAM cells and IM cells isolated from healthy pigs after initial and secondary treatment in vitro.
See this image and copyright information in PMC

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