Granoff D M, Kelsey S K, Bijlmer H A, Van Alphen L, Dankert J, Mandrell R E, Azmi F H, Scholten R J
Children's Hospital Oakland Research Institute, California 94609, USA.
Clin Diagn Lab Immunol. 1995 Sep;2(5):574-82. doi: 10.1128/cdli.2.5.574-582.1995.
We measured antibody responses to meningococcal serogroup B (MenB) polysaccharide (PS) by enzyme-linked immunosorbent assay (ELISA) in sera from 94 patients from The Netherlands with disease caused by Neisseria meningitidis group B. The patients ranged in age from 3 to 73 years (mean age, 18.8 years). In initial studies we showed that the binding of a panel of MenB PS-reactive human immunoglobulin M (IgM) paraproteins to biotinylated MenB PS bound to avidin-coated microtiter wells was inhibited > 90% by the addition of soluble MenB PS or encapsulated group B meningococci. In contrast, inhibition of IgM anti-MenB PS antibody-binding activity in many of the patient sera was less than 50% (range, 20 to 94%). These data suggested a high frequency of nonspecific binding in the patient sera. Therefore, all serum samples were assayed in replicate in the presence or absence of soluble MenB PS, and only the inhibitable fraction of the binding signal was used to calculate the anti-MenB PS antibody concentrations. In 17 control patients with meningococcal disease caused by serogroup A or C strains, there was no significant difference in the respective IgM or IgG anti-MenB PS antibody concentrations in paired acute- and convalescent-phase sera. In contrast, in patients with MenB disease, the geometric mean IgM anti-MenB PS antibody concentration increased from 3.9 units/ml in acute-phase serum to 10.5 units/ml in convalescent-phase serum (P < 0.001). The corresponding geometric mean IgG anti-MenB PS antibody titers were 1:27 and 1:36 (P < 0.05). There was only a weak relationship between age and the magnitude of the logarithm of the antibody concentrations in convalescent-phase sera (for IgM, r2 = 0.06 and P < 0.05; for IgG, r2 = 0.08 and P < 0.01). Our data indicate that precautions are needed to avoid nonspecificity in measuring serum antibody responses to MenB PS by ELISA. Furthermore, although this PS is thought to be a poor immunogen, patients as young as 3 years of age recovering from MenB disease demonstrate both ImG and IgG antibody responses in serum.
我们采用酶联免疫吸附测定(ELISA)法,检测了94例荷兰B群脑膜炎奈瑟菌病患者血清中针对B群脑膜炎球菌多糖(PS)的抗体反应。患者年龄在3至73岁之间(平均年龄18.8岁)。在初步研究中,我们发现,向包被抗生物素蛋白的微量滴定孔中结合的生物素化B群脑膜炎球菌PS添加可溶性B群脑膜炎球菌PS或包膜B群脑膜炎球菌后,一组B群脑膜炎球菌PS反应性人免疫球蛋白M(IgM)副蛋白与之的结合被抑制了90%以上。相比之下,许多患者血清中IgM抗B群脑膜炎球菌PS抗体结合活性的抑制率不到50%(范围为20%至94%)。这些数据表明患者血清中存在高频非特异性结合。因此,所有血清样本均在有或无可溶性B群脑膜炎球菌PS的情况下进行重复检测,并且仅使用结合信号的可抑制部分来计算抗B群脑膜炎球菌PS抗体浓度。在17例由A群或C群菌株引起的脑膜炎球菌病对照患者中,配对的急性期和恢复期血清中各自的IgM或IgG抗B群脑膜炎球菌PS抗体浓度没有显著差异。相比之下,在B群脑膜炎球菌病患者中,IgM抗B群脑膜炎球菌PS抗体几何平均浓度从急性期血清中的3.9单位/毫升增加到恢复期血清中的10.5单位/毫升(P<0.001)。相应的IgG抗B群脑膜炎球菌PS抗体几何平均滴度分别为1:27和1:36(P<0.05)。恢复期血清中抗体浓度对数的大小与年龄之间仅存在微弱的关系(对于IgM,r2=0.06,P<0.05;对于IgG,r2=0.08,P<0.01)。我们的数据表明,采用ELISA法检测血清中针对B群脑膜炎球菌PS的抗体反应时,需要采取预防措施以避免非特异性。此外,尽管这种PS被认为是一种弱免疫原,但从B群脑膜炎球菌病康复的3岁幼儿血清中也显示出IgM和IgG抗体反应。