Jarvis G A, Griffiss J M
Center for Immunochemistry, University of California, San Francisco.
J Immunol. 1989 Sep 1;143(5):1703-9.
We studied the effect of human IgA1, the predominant IgA subclass in serum, on C-mediated killing of Neisseria meningitidis. We purified monomeric IgA1 from normal human serum and tetravalent meningococcal polysaccharide vaccinate serum by using the following successive chromatographic steps: jacalin lectin affinity, Superose 12 FPLC gel filtration, Mono Q FPLC anion exchange, and anti-IgG affinity. SDS-PAGE, ELISA, and Western immunoblot analyses of the IgA1 detected no trace of contaminating IgG or IgM. IgA1 initiated partial or complete lysis (62 to 100%) of nine group C strains by using either normal, hypogammaglobulinemic, factor B-depleted, or properdin-deficient human serum as a C source, but IgA1 was unable to effect killing in serum chelated with 10 mM MgCl2 and 10 mM EGTA. Lytic activity was dependent on the group C strain and the source of the IgA1; neither IgA1 preparation was bactericidal for all nine strains. Removal of the Fc portion of IgA1 with pepsin completely abolished bactericidal activity. We purified and radiolabeled C component C3, and found that IgA1 did not increase C3 deposition. With the use of a group C polysaccharide ELISA, we found that the vaccinate IgA1 had a high titer of group C polysaccharide antibody, whereas the IgA1 purified from normal human serum had no detectable group C polysaccharide specificity. Absorption of the vaccinate IgA1 with alum-bound group C polysaccharide did not affect the killing of a sensitive strain, but it did potentiate the killing of a previously resistant strain. Western immunoblots of whole cell lysates, outer membrane complex, and purified lipooligosaccharide showed that the bactericidal IgA1 was specific for several outer membrane proteins. Four of the proteins recognized by both IgA1 preparations had apparent Mr of 29, 42, 66, and 74 kDa. We conclude that IgA1, when bound to specific outer membrane proteins, can initiate lysis of group C meningococci via the classical C pathway, and that initiation of lysis is an Fc-dependent event which occurs without an increase in C3 deposition.
我们研究了血清中主要的IgA亚类——人IgA1对补体(C)介导的脑膜炎奈瑟菌杀伤作用的影响。我们通过以下连续的色谱步骤从正常人血清和四价脑膜炎球菌多糖疫苗接种血清中纯化单体IgA1:jacalin凝集素亲和层析、Superose 12 FPLC凝胶过滤、Mono Q FPLC阴离子交换和抗IgG亲和层析。对纯化的IgA1进行SDS-PAGE、ELISA和Western免疫印迹分析,未检测到污染的IgG或IgM痕迹。IgA1以正常、低丙种球蛋白血症、因子B缺陷或备解素缺陷的人血清作为补体来源,可引发9株C群菌株的部分或完全裂解(62%至100%),但IgA1在与10 mM MgCl2和10 mM EGTA螯合的血清中无法发挥杀伤作用。裂解活性取决于C群菌株和IgA1的来源;两种IgA1制剂均不能对所有9株菌株产生杀菌作用。用胃蛋白酶去除IgA1的Fc部分完全消除了杀菌活性。我们纯化并放射性标记了补体成分C3,发现IgA1不会增加C3沉积。通过使用C群多糖ELISA,我们发现疫苗接种来源的IgA1具有高滴度的C群多糖抗体,而从正常人血清中纯化的IgA1未检测到C群多糖特异性。用明矾结合的C群多糖吸收疫苗接种来源的IgA1不影响对敏感菌株的杀伤,但确实增强了对先前耐药菌株的杀伤。对全细胞裂解物、外膜复合物和纯化的脂寡糖进行Western免疫印迹分析表明,具有杀菌作用的IgA1对几种外膜蛋白具有特异性。两种IgA1制剂识别的四种蛋白质的表观分子量分别为29、42、66和74 kDa。我们得出结论,IgA1与特定外膜蛋白结合时,可通过经典补体途径引发C群脑膜炎球菌的裂解,并且裂解的起始是一个Fc依赖性事件,发生时C3沉积不增加。