Mononuclear phagocytes (MP) are cells of nonspecific immunity, playing an essential role in defense against bacterial pathogens. Although various MP subpopulations have been described in the pig, relations among these populations in vivo are unknown to date. The present study was aimed at describing porcine MP subpopulations infiltrating inflamed tissue of pigs under in vivo conditions. Actinobacillus pleuropneumoniae (APP) infection was used to induce an inflammatory response. CD172alpha, CD14, CD163, MHCII and CD203alpha cell surface molecules were used to identify MP by flow cytometry. Changes in MP subpopulations in the peripheral blood (PB) and bone marrow (BM) compartments along with the analysis of MP appearing in the inflamed lungs were assessed to elucidate the possible origin and maturation stages of the infiltrating MP. The MP population migrating to the inflamed lungs was phenotype CD14+ CD163+ CD203alpha+/- MHCII+/-. Concomitantly, after APP infection there was an increase in the PB MP CD14+ CD163+ CD203alpha- MHC II- population, suggesting that these cells give rise to inflammatory monocytes/macrophages. The CD203alpha and MHCII molecules appear on these cells after leaving the PB. In healthy animals, the BM MP precursors were represented by CD14- CD163- cells maturing directly into CD14+ CD163- that were then released into the PB. After infection, an altered maturation pathway of MP precursors appeared, represented by CD14- CD163- CD203alpha- MHCII- MP directly switching into CD14+ CD163+ CD203alpha- MHCII- MP. In conclusion, two different MP maturation pathways were suggested in pigs. The use of these pathways differs under inflammatory and noninflammatory conditions.

Department of Immunology Veterinary Research Institute Hudcova 70 621 00 Brno Czech Republic

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Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines