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Transcriptomic establish of pig macrophage polarization signatures and potential applications for pig health and disease

ABSTRACT: Macrophages are the main effector cells of inflammatory response and play an important role in the initiation and regression of inflammation. Macrophages are derived from monocytes which are differentiated into M1 macrophages (classically activated macrophages) or M2 macrophages (selectively activated macrophages) in response to different external stimuli, distinguished by the differential expression of cytokines, cell surface markers, and chemokines. In vitro macrophages are crucial models for understanding the regulatory mechanism of immune response and the establishment of infection models, ultimately, for the diagnosis or treatment of a variety of diseases. Porcine is the most important agricultural animal and one of the most valuable animal models for preclinical studies, but no unified method for porcine macrophage isolation and differentiation at present, neither a systematic study compared the porcine macrophage obtained by different methods. In this study, we used two strategies to induce porcine BMDM into M1 and two strategies to induce porcine BMDM into M2 macrophages. Subsequently, we compared the transcriptomic profile of BMDM treated with different induction methods by RNA-seq. The DEG analysis showed that porcine M1 macrophage and porcine M2 macrophage obtained have consistent gene signatures as mouse and human macrophages, respectively. We also observed differences in transcriptional levels in macrophages induced by different methods. We obtained the transcriptome sequencing data of macrophages infected by different pathogens from GEO and analyzed them quantitatively. We performed GSEA analysis on macrophages infected with different pathogens and porcine bone marrow-derived macrophages (BMDM). We found that the enrichment pathways of macrophages obtained by different induction methods and macrophages infected by different pathogens were different. Our study provided a framework to guide the interrogation of macrophage phenotypes in the context of health and disease. The approach described here could be used to propose new biomarkers for diagnosis in diverse clinical settings including porcine reproductive and respiratory syndrome virus (PRRSV), African swine fever virus (ASFV), and Toxoplasma gondii.

ORGANISM(S): Sus scrofa

PROVIDER: GSE202115 | GEO | 2025/05/03

REPOSITORIES: GEO

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Project description:Recent studies suggest the presence of both âclassically activatedâ M1 and âalternatively activatedâ M2 macrophages in human atherosclerotic tissue, yet due to the lack of validated markers the reported localization patterns of these macrophage phenotypes within plaques are ambiguous. In the present study, we searched for markers that indisputably can identify differentiated M1 and M2 macrophages independently of stimuli that affect the activation status of the two subpopulations. We used these validated markers to assess the presence of M1 and M2 macrophages in different zones of human carotid artery atherosclerotic plaques obtained from 12 patients. Using microarray and qPCR technology we show that the frequently used macrophage subpopulation markers MCP-1 and CD206 do not discriminate between M1 and M2 macrophages. However, we confirm the subtype specificity of the classical M2 marker CD163 and we report that the genes INHBA and DSP (both M1) and SEPP1 and MARCKS (both M2) are highly suitable for macrophage phenotyping. mRNA expression of the pan-macrophage marker CD68 in the shoulder zones of the plaques and in adjacent tissue segments correlated positively with mRNA expression levels of SEPP1, MARCKS and CD163 (r=0.86, 0.94 and 0.96, and r= 0.86, 0.98 and 0.69, respectively) but not with the expression of the M1 markers DSP and INHBA. In contrast, mRNA expression of CD68 in the core of the plaques correlated positively with expression of DSP and INHBA (r=0.73 and 0.49) but not with SEPP1, MARCKS and CD163. These findings suggest that M1 macrophages predominate in the core of human carotid atherosclerotic plaques while M2 macrophages prevail at the periphery of the plaque. Keywords: Expression profiling by array Monocytes from healthy volunteers were differentiated into M1 and M2 macrophages by incubation with granulocyte-macrophage colony-stimulating factor (GM-CSF) or macrophage colony-stimulating factor (M-CSF), respectively. After 5 days cells were exposed to oxidized LDL. Total RNA was isolated and subjected to gene expression profiling.

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Transcriptomic establish of pig macrophage polarization signatures and potential applications for pig health and disease

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