Rot A, Henderson L E, Leonard E J
J Leukoc Biol. 1986 Jul;40(1):43-53. doi: 10.1002/jlb.40.1.43.
Products of bacteria are potent chemoattractants for mammalian leukocytes. Several reports suggest that these attractants are small peptides. We compared the properties of culture fluids of Staphylococcus aureus with fMet-Leu-Phe, considered a prototype of bacterial attractant. Chemotactic activity for human monocytes of Staph. aureus culture filtrates was determined in multiwell chemotaxis chambers. At optimal concentrations, the filtrate attracted almost twice as many monocytes as fMet-Leu-Phe (53 +/- 5% of input number compared with 30 +/- 3%, in a series of ten experiments). Gel-filtration characteristics and susceptibility to proteolytic digestion suggested that chemotactic activity was due to peptides with a molecular size range of 500-2,000 daltons. Reverse-phase high-pressure liquid chromatography (HPLC) of unfractionated filtrate revealed nine peaks of chemotactic activity, most of which was in five of the peaks. One peak accounted for 40% of total activity. Individual peaks, like the unfractionated material, were capable of attracting about twice as many monocytes as the optimal concentration of fMet-Leu-Phe. Quantitative bioassay of the HPLC peaks showed that only 5% of the total Staph. aureus chemotactic activity eluted in the position of fMet-Leu-Phe. This is in contrast to the report that fMet-Leu-Phe accounted for 70% of neutrophil lysosomal enzyme-releasing activity of Escherichia coli culture fluid. In summary, chemotactic activity for monocytes of Staph. aureus culture fluid is due to peptides other than fMet-Leu-Phe; these peptides recruit a higher percentage of monocytes than fMet-Leu-Phe.
细菌产物是哺乳动物白细胞强有力的化学引诱剂。几份报告表明这些引诱剂是小肽。我们将金黄色葡萄球菌培养液的特性与被视为细菌引诱剂原型的甲酰甲硫氨酸-亮氨酸-苯丙氨酸(fMet-Leu-Phe)进行了比较。在多孔趋化性小室中测定了金黄色葡萄球菌培养滤液对人单核细胞的趋化活性。在最佳浓度下,滤液吸引的单核细胞数量几乎是fMet-Leu-Phe的两倍(在一系列十次实验中,分别为输入细胞数的53±5%和30±3%)。凝胶过滤特性和对蛋白水解消化的敏感性表明趋化活性归因于分子大小在500 - 2000道尔顿范围内的肽。未分级滤液的反相高压液相色谱(HPLC)显示出九个趋化活性峰,其中大部分活性集中在五个峰中。一个峰占总活性的40%。与未分级的物质一样,各个峰吸引的单核细胞数量约为fMet-Leu-Phe最佳浓度时的两倍。对HPLC峰的定量生物测定表明,金黄色葡萄球菌的总趋化活性中只有5%在fMet-Leu-Phe的位置洗脱。这与一份报告形成对比,该报告称fMet-Leu-Phe占大肠杆菌培养液中性粒细胞溶酶体酶释放活性的70%。总之,金黄色葡萄球菌培养液对单核细胞的趋化活性归因于fMet-Leu-Phe以外的肽;这些肽招募的单核细胞百分比高于fMet-Leu-Phe。