猪繁殖与呼吸综合征病毒-脂多糖刺激对中性粒细胞黏附猪肺微血管内皮细胞的影响

doi:10.16431/j.cnki.1671-7236.2022.02.008

Abstract

Abstract: 【Objective】 This study was aimed to study the effects of Porcine reproductive and respiratory syndrome virus (PRRSV) alone and in combination with lipopolysaccharide (LPS) stimulation on lung recruitment neutrophil (Neu), and the role of porcine lung microvascular endothelial cells (MVECs).【Method】 The primary MVECs were isolated and cultured by type Ⅱ collagenase digestion and differential adhesion method.Neu was isolated from porcine peripheral blood by density gradient centrifugation.After stimulating Neu and MVECs with PRRSV HN strain or PRRSV-LPS, Neu was added to the culture plate with MVECs, incubated for 1 h, fixed with 4% paraformaldehyde, stained with Wright's staining, and the number of Neu adhered in each group was observed, counted and analyzed.The expression of P-selectin and E-selectin in MVECs was detected by immunocytochemical staining.The quantitative changes of Neu adhesion to MVECs stimulated by PRRSV HN strain or PRRSV-LPS were measured and analyzed by rose bengal solution staining when MVECs were blocked by P-selectin and E-selectin antibodies.【Result】 The isolated and cultured MVECs were CD34 immunofluorescence positive, and the positive rate was about 92%.When PRRSV HN strain was stimulated for 18 h, the number of Neu adhered to MVECs increased significantly (P < 0.05).When stimulated with PRRSV-LPS, the number of adherent Neu was significantly higher than that stimulated with LPS alone (P < 0.05).When PRRSV or PRRSV-LPS stimulated Neu and MVECs, the increase in the number of adherent Neu was more obvious than that only stimulated MVECs or Neu.Immunocytochemical staining showed that MVECs strongly expressed P-selectin and E-selectin.When P-selectin and E-selectin antibodies blocked MVECs, the increase of Neu adhesion induced by PRRSV or PRRSV-LPS decreased in varying degrees, and there was a significant difference when E-selectin blocked MVECs (P < 0.05).【Conclusion】 PRRSV infection could not only promote the adhesion between Neu and MVECs, but also increase the number of Neu adhered during subsequent LPS stimulation.E-selectin expressed by MVECs was one of the important molecules mediating the increase of Neu adhesion caused by PRRSV.

Key words: Porcine reproductive and respiratory syndrome virus (PRRSV); neutrophil (Neu); microvascular endothelial cells (MVECs); adhesion; selectin

CLC Number:

S852.65⁺1

WANG Zhaoli, YANG Siyu, WU Yanmei, SONG Xiaoxiao, MU Xiang, ZHANG Tao. Effects of PRRSV-LPS Treatment on Neutrophil Adhesion to Porcine Pulmonary Microvascular Endothelial Cells[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 471-479.

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Effects of PRRSV-LPS Treatment on Neutrophil Adhesion to Porcine Pulmonary Microvascular Endothelial Cells

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