Rice P A, Kasper D L
J Clin Invest. 1977 Nov;60(5):1149-58. doi: 10.1172/JCI108867.
The role of gonococcal antigens in serum bactericidal activity was determined for an isolate of Neisseria gonorrhoeae from a patient with disseminated gonococcal infection (DGI). Gonococcal outer membranes were purified by differential ultracentrifugation of sheared organisms treated with EDTA. The membranes were solubilized in an endotoxin-disaggregating buffer, and the proteins were separated from the endotoxin by molecular sieve chromatography. Chemical characterization of the endotoxin from the DGI strain revealed the presence of heptose (7.8% of carbohydrate composition) and 2-keto-3-deoxyoctonate (6.1%, wt/wt) in concentrations similar to rough endotoxins of gram-negative enteric bacteria. Dermal Schwartzman reactions were positive for this endotoxin (4 mug) and the corresponding outer membrane (139 mug), but negative for the protein fraction (>500 mug). The patient's whole serum or the IgG fraction, each with complement, reduced the number of the infecting organisms by greater than 1 log(10) in a bactericidal assay. Outer membrane and its protein and endotoxin fractions (0.8-500 mug) from the DGI strain were separately preincubated with the patient's convalescent serum and specific inhibition of bactericidal activity occurred with the endotoxin fraction (25 mug) and the outer membrane (100 mug); the protein (500 mug) exhibited no inhibition. Similar results were obtained by inhibiting the bactericidal activity of rabbit antiserum, prepared by intravenous inoculation of an isolate from a patient with pelvic inflammatory disease, with antigen purified from that strain. That this was specific immune inhibition and not anticomplementary activity was shown by the failure of these antigens to inhibit other complement-dependent bactericidal systems.
针对一名患有播散性淋球菌感染(DGI)患者的淋病奈瑟菌分离株,测定了淋球菌抗原在血清杀菌活性中的作用。通过对用EDTA处理的剪切菌进行差速超速离心来纯化淋球菌外膜。将膜溶解在内毒素分解缓冲液中,并通过分子筛色谱法将蛋白质与内毒素分离。对DGI菌株的内毒素进行化学表征,结果显示庚糖(占碳水化合物组成的7.8%)和2-酮-3-脱氧辛酸(6.1%,重量/重量)的浓度与革兰氏阴性肠道细菌的粗糙内毒素相似。该内毒素(4微克)和相应的外膜(139微克)的皮肤施瓦茨曼反应呈阳性,但蛋白质部分(>500微克)呈阴性。在杀菌试验中,患者的全血清或IgG组分(均含补体)使感染菌数量减少超过1个对数(10)。将DGI菌株的外膜及其蛋白质和内毒素组分(0.8 - 500微克)分别与患者的恢复期血清预孵育,内毒素组分(25微克)和外膜(100微克)出现杀菌活性的特异性抑制;蛋白质(500微克)未表现出抑制作用。用从一名盆腔炎患者分离株制备的兔抗血清进行试验,用从该菌株纯化的抗原抑制其杀菌活性,也得到了类似结果。这些抗原未能抑制其他补体依赖性杀菌系统,表明这是特异性免疫抑制而非抗补体活性。