Schneider H, Hammack C A, Apicella M A, Griffiss J M
Department of Bacterial Diseases, Walter Reed Army Institute of Research, Washington, D.C. 20307.
Infect Immun. 1988 Apr;56(4):942-6. doi: 10.1128/iai.56.4.942-946.1988.
We assessed variation in the expression of lipooligosaccharide (LOS) components and their epitopes within populations of a strain of Neisseria gonorrhoeae by using the monoclonal antibodies (MAbs) O6B4 and 3F11 and immunoenzymatic, immuno-colloidal gold electron microscopic, and sodium dodecyl sulfate-polyacrylamide gel electrophoretic procedures. Wild-type organisms varied in binding of both MAbs. We used the intensity of immunoenzymatic colony blot color to distinguish four binding variants for each MAb: red (R), pink (P), and colorless (nonreactive [N]) and an N back to R (N-R) revertant. R to P to R and R to N to R variation occurred at frequencies of 0.2% and 0.02%, respectively. The electrophoretic LOS profiles and MAb immunoblot patterns of the R, P, and N-R variants were the same as those of the wild type. LOSs of the N variants, in contrast, were of lower Mr, bound neither 3F11 nor O6B4 MAb, and contained as their major component the 3.6-kilodalton LOS that bears the L8LOS epitope of N. meningitidis. Results of immunoelectron microscopic studies were consistent with LOS binding patterns. Large number of colloidal gold particles were deposited about both R and P variants, distally from R organisms, but proximally from P organisms. N variant organisms, like their LOS, bound neither of the MAbs. N-R variant organisms were like the wild type in that they showed much variation in the amounts of MAb they bound.
我们通过使用单克隆抗体(MAb)O6B4和3F11以及免疫酶法、免疫胶体金电子显微镜法和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法,评估了淋病奈瑟菌菌株群体中脂寡糖(LOS)成分及其表位表达的变异情况。野生型菌株在两种单克隆抗体的结合上存在差异。我们利用免疫酶菌落印迹颜色的强度,为每种单克隆抗体区分出四种结合变异体:红色(R)、粉色(P)和无色(无反应性[N])以及从N变回R(N-R)的回复体。从R到P再到R以及从R到N再到R的变异发生率分别为0.2%和0.02%。R、P和N-R变异体的电泳LOS图谱和单克隆抗体免疫印迹模式与野生型相同。相比之下,N变异体的LOS分子量较低,既不与3F11单克隆抗体结合,也不与O6B4单克隆抗体结合,其主要成分是带有脑膜炎奈瑟菌L8LOS表位的3.6千道尔顿LOS。免疫电子显微镜研究结果与LOS结合模式一致。大量胶体金颗粒沉积在R和P变异体周围,远离R型菌株,但靠近P型菌株。N变异体菌株与其LOS一样,不与任何一种单克隆抗体结合。N-R变异体菌株与野生型相似,它们在结合的单克隆抗体数量上表现出很大差异。