Jarvis G A, Vedros N A
Infect Immun. 1987 Jan;55(1):174-80. doi: 10.1128/iai.55.1.174-180.1987.
The effect of meningococcal cell-associated sialic acid on activation of the human alternative complement pathway was examined by using a quantitative fluorescence immunoassay to assess alternative pathway-mediated C3 binding to a group B strain of Neisseria meningitidis from which graded amounts of sialic acid had been removed with neuraminidase. Using human serum absorbed with strain B16B6 (B:2a:L2,3) and chelated with 10 mM MgCl2 and 10 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, we found an increase in the amount of C3 bound by enzymatically desialylated B16B6 organisms over the amount bound by fully sialylated organisms. This increase was proportional to the amount of sialic acid cleaved from the bacteria. Enhanced C3 binding was accompanied by an increase in factor B deposition. A sialic acid-deficient mutant of strain B16B6, designated 2T4-1, bound C3 via the alternative pathway at a level equivalent to that bound by wild-type meningococci from which 88% of the sialic acid had been removed. Strain B16B6 was resistant to the alternative pathway-mediated bactericidal activity of both absorbed and hypogammaglobulinemic human sera, whereas noncapsular variant 2T4-1 was sensitive to these sera. The addition of purified immune immunoglobulin M (IgM) and IgG significantly increased the alternative pathway-mediated killing of strain B16B6 organisms. IgM mediated increased bactericidal activity without an increase in C3 or factor B deposition. In contrast, the IgG-mediated killing was associated with increased binding of C3 and factor B to the organisms. Absorption studies showed that the IgM bound to the sialic acid capsule, whereas the IgG bound to noncapsular surface antigens. We conclude from these results that the group B meningococcal sialic acid capsule inhibits activation of the alternative pathway in the nonimmune host and that both IgM and IgG, although specific for different surface antigens, are capable of augmenting the alternative pathway-mediated killing of group B meningococci.
通过定量荧光免疫测定法,检测了脑膜炎球菌细胞相关唾液酸对人替代补体途径激活的影响,以评估替代途径介导的C3与B群脑膜炎奈瑟菌的结合情况,该菌已用神经氨酸酶去除了不同量的唾液酸。使用经B16B6菌株(B:2a:L2,3)吸收并与10 mM MgCl2和10 mM乙二醇双(β-氨基乙醚)-N,N,N',N'-四乙酸螯合的人血清,我们发现,经酶解去唾液酸化的B16B6菌结合的C3量比完全唾液酸化的菌结合的C3量增加。这种增加与从细菌中裂解的唾液酸量成比例。C3结合增强伴随着B因子沉积增加。B16B6菌株的一个唾液酸缺陷突变体,命名为2T4-1,通过替代途径结合C3的水平与已去除88%唾液酸的野生型脑膜炎球菌相当。B16B6菌株对吸收后的人血清和低丙种球蛋白血症人血清的替代途径介导的杀菌活性具有抗性,而非荚膜变体2T4-1对这些血清敏感。添加纯化的免疫免疫球蛋白M(IgM)和IgG显著增加了替代途径介导的B16B6菌株的杀伤作用。IgM介导杀菌活性增加,而C3或B因子沉积没有增加。相比之下,IgG介导的杀伤与C3和B因子与细菌的结合增加有关。吸收研究表明,IgM结合到唾液酸荚膜上,而IgG结合到非荚膜表面抗原上。我们从这些结果得出结论,B群脑膜炎球菌唾液酸荚膜在非免疫宿主中抑制替代途径的激活,并且IgM和IgG虽然对不同的表面抗原具有特异性,但都能够增强替代途径介导的B群脑膜炎球菌的杀伤作用。