Walter Michaela R V, Morck Douglas W
Department of Biological Sciences, University of Calgary, AB, Canada.
Am J Vet Res. 2002 May;63(5):757-62. doi: 10.2460/ajvr.2002.63.757.
To examine the host response toward Porphyromonas levii, by evaluating chemotaxis, phagocytosis, and oxidative burst of bovine macrophages in vitro.
Cultured bovine macrophages obtained from monocytes harvested from blood samples of 15 Holstein steers. Porphyromonas levii was isolated from the foot rot lesion of an acutely affected feedlot steer.
Monocytes were cultured for macrophage differentiation over 7 days. Porphyromonas levii was cultured in strict anaerobic conditions for experimentation. Chemotaxis was evaluated by quantifying macrophage migration toward P. levii in Boyden chambers. Phagocytosis was assessed by quantification of macrophages engulfing P. levii following incubation with or without anti-P. levii serum or purified IgG. Oxidative burst was measured by use of the nitroblue tetrazolium reduction assay.
Chemotaxis toward P. levii was not significantly different from control values at any of the tested bacterial concentrations. Phagocytosis of P. levii was approximately 10% at a 10:1 bacterium to macrophage ratio and did not change significantly over time. When higher proportions of P. levii were tested for phagocytosis, the 1,000:1 bacterium to macrophage ratio had a significant increase, compared with the 10:1 test group. Opsonization of P. levii with high-titeranti-P. levii serum or anti-P. levii IgG produced a significant increase in macrophage phagocytosis. Oxidative production significantly increased compared with control in the 1,000:1 test group only.
Porphyromonas levii may evade host detection by decreased chemotaxis, phagocytosis, and oxidative burst by macrophages. Acquired immunity may be beneficial for clearance of P. levii in foot rot lesions in cattle.
通过体外评估牛巨噬细胞的趋化性、吞噬作用和氧化爆发,研究宿主对牙龈卟啉单胞菌的反应。
从15头荷斯坦公牛血液样本中采集单核细胞培养获得的牛巨噬细胞。牙龈卟啉单胞菌从一头急性发病饲养场公牛的腐蹄病病变中分离得到。
单核细胞培养7天以分化为巨噬细胞。牙龈卟啉单胞菌在严格厌氧条件下培养用于实验。通过在博伊登小室中量化巨噬细胞向牙龈卟啉单胞菌的迁移来评估趋化性。通过量化巨噬细胞在与抗牙龈卟啉单胞菌血清或纯化IgG孵育后吞噬牙龈卟啉单胞菌的情况来评估吞噬作用。使用硝基蓝四氮唑还原试验测量氧化爆发。
在任何测试的细菌浓度下,对牙龈卟啉单胞菌的趋化性与对照值均无显著差异。在细菌与巨噬细胞比例为10:1时,牙龈卟啉单胞菌的吞噬率约为10%,且随时间无显著变化。当测试更高比例的牙龈卟啉单胞菌进行吞噬作用时,与10:1测试组相比,1000:1的细菌与巨噬细胞比例有显著增加。用高滴度抗牙龈卟啉单胞菌血清或抗牙龈卟啉单胞菌IgG调理牙龈卟啉单胞菌可显著增加巨噬细胞吞噬作用。仅在1000:1测试组中,氧化产物与对照相比显著增加。
牙龈卟啉单胞菌可能通过降低巨噬细胞的趋化性、吞噬作用和氧化爆发来逃避宿主检测。获得性免疫可能有利于清除牛腐蹄病病变中的牙龈卟啉单胞菌。