Polarization Heterogeneity and Reversibility of Porcine (Sus scrofa) Alveolar Macrophages
WU Ya-Lan1,2, WANG Zhen-Yue1, WANG Dao-Yuan1,2, ZHANG Wei1,2, LI Xin-Yun1,2,3, ZHAO Shu-Hong1,2,3, ZHU Meng-Jin1,2,3,*
1 College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China; 2 Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, China; 3 Collaborative Innovation Center for Healthy Pig Breeding, Wuhan 430070, China
Abstract：Porcine alveolar macrophages (PAMs) are important members of the body's first line of defense in innate immunity. Their strong plasticity provides a significant cell model for investigating the disease resistance mechanism of pigs (Sus scrofa). In order to establish different polarized isoforms of porcine alveolar macrophages and to compare the polarization heterogeneity and reversibility of PAMs and 3D4/21 cells, PAMs isolated from 30-day-old healthy Yorkshire piglets and commercialized 3D4/21 cell line perserved by laboratory were used as experimental materials in this study. A combination of lipopolysaccharide (LPS) with interferon gamma (IFN-γ), interleukin 4 (IL-4) were used to induce the polarization of PAMs and 3D4/21 cells to construct M1 and M2 cell models, respectively. PAMs and 3D4/21 cells without polarization were used as the control group (M0 type). PCR and qRT-PCR relative quantitative results showed that the M1-type marker genes IL-1β, CXC chemokine ligand 10 (CXCL10), tumour necrosis factor (TNF), IL-6, and IL-12B were significantly more highly expressed in PAMs and 3D4/21 cell M1 types than M0 and M2 types (P<0.05); M2-type marker genes arginase-1 (Arg1), peroxisome proliferators-activated receptor γ (PPARγ), IL-10 and chitinase 3-like 3 (Chi3L3 or Ym-1) were significantly more highly expressed in M2 type than in M0 and M1 types in the 3D4/21 cell (P<0.05). In PAMs, the expression levels of Arg1, PPARγ and Ym-1 in M0 and M2 were significantly higher than that of in M1 (P<0.05). Western blot detections of Arg1 protein in the 2 types of cells were consistent with the relative quantitative results of qRT-PCR. At the same time, the reversibility test results of PAMs and 3D4/21 cell polarization showed that the reversibility of PAMs polarization was stronger than that of 3D4/21 cell, and the ability of PAMs to switch from M2 to M1 was stronger than from M1 to M2 type conversion capability. This study provides selection materials and basic data for further exploration of porcine alveolar macrophage polarization, immune mechanisms, and pig disease-resistant breeding.
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