Drogari-Apiranthitou M, Kuijper E J, Dekker N, Dankert J
Department of Medical Microbiology, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands.
Infect Immun. 2002 Jul;70(7):3752-8. doi: 10.1128/IAI.70.7.3752-3758.2002.
Encapsulated meningococci are complement sensitive only in the presence of bactericidal antibodies by yet-unexplored mechanisms. The objective of this study was to investigate the involvement of major bacterial surface constituents on complement activation and membrane attack complex (MAC) formation on serogroup B meningococci in the presence or absence of antibody-dependent serum bactericidal activity (SBA). The strains used were the encapsulated H44/76, five of its variants differing in capsulation and expression of the class 1 porin (PorA), and its lipopolysaccharide (LPS)-deficient isogenic mutant (LPS(-)) pLAK33. Two normal sera, one with high SBA (SBA(+)) and one with no bactericidal activity (SBA(-)) against H44/76 as well as an a-gamma-globulinemic serum were used for sensibilization of the bacteria. C3b and iC3b deposition on H44/76, its unencapsulated variant v24, and pLAK33 was similar in SBA(+) and SBA(-) serum, and no difference was present between the strains. MAC deposition on H44/76 was higher in SBA(+) serum than in SBA(-) serum and the a-gamma-globulinemic serum. The amounts of C3b on H44/76, v24, and pLAK33 in the a-gamma-globulinemic serum were also not different, indicating immunoglobulin G (IgG)- and LPS-independent complement activation. H44/76 PorA(+) and its PorA(-) variant and the v24 PorA(+) and its PorA(-) variant incubated in SBA(-) serum induced comparable amounts of MAC, despite their different serum sensitivities. Complement formation on the surface of the bacteria occurred almost exclusively via the classical pathway, but the considerable amounts of Bb measured in the serum indicated alternative pathway activation in the fluid phase. We conclude that complement deposition on meningococci is, for the most part, independent of classical pathway IgG and is not influenced by the presence of PorA or LPS on the meningococcal surface. Addition of an anti-PorA chimeric antibody to the nonbactericidal normal serum, while promoting a dose-related bacterial lysis, did not influence the amounts of C3b, iC3b, and MAC formed on the bacterial surface. These findings support the hypothesis that proper MAC insertion rather than the quantity of MAC formed on the bacterial surface is of importance for efficient lysis of meningococci.
被膜包裹的脑膜炎球菌仅在存在杀菌抗体时通过尚未明确的机制对补体敏感。本研究的目的是调查在存在或不存在抗体依赖性血清杀菌活性(SBA)的情况下,主要细菌表面成分对B群脑膜炎球菌补体激活和膜攻击复合物(MAC)形成的影响。所用菌株为被膜包裹的H44/76、其5个在被膜形成和1类孔蛋白(PorA)表达上不同的变体,以及其脂多糖(LPS)缺陷的同基因突变体(LPS(-))pLAK33。使用两份正常血清,一份对H44/76具有高SBA(SBA(+)),另一份对H44/76无杀菌活性(SBA(-)),以及一份无丙种球蛋白血症血清对细菌进行致敏。在SBA(+)和SBA(-)血清中,H44/76、其无被膜变体v24和pLAK33上C3b和iC3b的沉积相似,且各菌株之间无差异。SBA(+)血清中H44/76上的MAC沉积高于SBA(-)血清和无丙种球蛋白血症血清。无丙种球蛋白血症血清中H44/76、v24和pLAK33上的C3b量也无差异,表明补体激活不依赖于免疫球蛋白G(IgG)和LPS。尽管H44/76 PorA(+)及其PorA(-)变体以及v24 PorA(+)及其PorA(-)变体在血清敏感性上不同,但在SBA(-)血清中孵育时诱导的MAC量相当。细菌表面的补体形成几乎完全通过经典途径发生,但血清中检测到的大量Bb表明液相中替代途径被激活。我们得出结论,脑膜炎球菌上的补体沉积在很大程度上不依赖于经典途径IgG,且不受脑膜炎球菌表面PorA或LPS存在的影响。向非杀菌正常血清中添加抗PorA嵌合抗体,虽然促进了剂量相关的细菌裂解,但并未影响细菌表面形成的C3b、iC3b和MAC量。这些发现支持这样的假设,即适当的MAC插入而非细菌表面形成的MAC数量对于脑膜炎球菌的有效裂解很重要。