Generation of chemotactic activity in serum by Haemophilus influenzae type b.
Generation of chemotactic activity in serum by Haemophilus influenzae type b.
复制标题
b 型流感嗜血杆菌在血清中产生趋化活性。
DOI：
10.1128/iai.43.2.593-599.1984
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发表时间：
1984
影响因子：
3.1
通讯作者：
Anderson,DC
中科院分区：
文献类型：
--
作者：
Tosi,MF;Kaplan,SL;Mason,EO;Buffone,GJ;Anderson,DC
进行研究以表征B型流感嗜血杆菌（HiTb）在血清中产生的趋化活性或不依赖于血清进行阐述。中性粒细胞聚集测定法，Sephadex G-75凝胶色谱法，和抗C5中和研究被用来证明，补体片段C5 a代表主要的趋化性部分，来自HiTb-血清相互作用。HiTb独立地阐述了最小的趋化活性。通过嗜中性粒细胞在趋化性、形状变化和聚集测定中的反应测量的HiTb产生的最大C5 a需要针对荚膜多糖、多聚核糖基核糖醇磷酸（PRP）的特异性抗体。与低丙种球蛋白血症血清相比，含有高滴度抗PRP（通过酶联免疫吸附试验测定）的合并正常人血清中产生的C5 a显著更多。此外，在用纯化的高滴度免疫球蛋白G、超免疫兔血清或热灭活的正常人血清重建的低丙种球蛋白血症血清中产生的C5 a与在正常人血清中产生的C5 a相当。与完整HiTb相比，PRP对抗体的吸收显示出非PRP定向抗体的贡献。如使用C4缺陷豚鼠血清所示，在非免疫血清中通过补体旁路途径产生C5 a，特异性抗PRP促进补体旁路途径的激活。C5 a在测试血清中的产生与其对HiTb的调理活性成比例，如通过鲁米诺-化学发光测定所评估的。在HiTb脑膜炎急性期获得的13例儿科患者血清样本中观察到C5 a活性总体水平较低，在这些患者中未观察到与白细胞聚集综合征一致的肺部症状或影像学异常。
Studies were performed to characterize chemotactic activity generated by Haemophilus influenzae type b (HiTb) in serum or elaborated independent of serum. Neutrophil aggregometry, Sephadex G-75 gel chromatography, and anti-C5 neutralization studies were used to demonstrate that the complement fragment C5a represented the major chemotactic moiety derived from HiTb-serum interactions. HiTb elaborated minimal chemotactic activity independently. Maximal C5a generation by HiTb as measured by neutrophil response in chemotaxis, shape change, and aggregation assays required specific antibody to the capsular polysaccharide, polyribosyl ribitol phosphate (PRP). Significantly more C5a was generated in pooled normal human serum containing high titers of anti-PRP (determined by an enzyme-linked immunosorbent assay) than in hypogammaglobulinemic serum. Furthermore, C5a generated in hypogammaglobulinemic serum reconstituted with purified high-titer immunoglobulin G, hyperimmune rabbit serum or heat-inactivated normal human serum was comparable to that generated in normal human serum. Absorption of antibody with PRP versus whole HiTb showed a contribution by non-PRP-directed antibody. As shown with the use of C4-deficient guinea pig serum, C5a generation occurred via the alternative complement pathway in nonimmune serum, and activation of the alternative complement pathway was facilitated by specific anti-PRP. C5a generation in test sera was proportional to its opsonic activity for HiTb as assessed by a luminol-chemiluminescence assay. Overall low levels of C5a activity were observed in 13 pediatric patient serum samples obtained during the acute phase of HiTb meningitis, and no pulmonary symptoms or radiographic abnormalities consistent with a leukocyte aggregation syndrome were observed in these patients.