ROLE OF NON-BACTERICIDAL ANTIBODIES IN PROTECTION AGAINST MENINGOCOCCAL DISEASE In the 1960s, Goldschneider et al provided compelling evidence that the

ability of serum antibodies together with complement to kill meningococci conferred protection against developing meningococcal disease. Today, a positive serum BA titer remains the only widely accepted correlate of protection against meningococcal disease. What is more controversial is whether persons with serum BA titers <1:4 also can be protected against developing meningococcal disease. We measured BA against three group B meningococcal strains in stored sera from 48 healthy adults, and in whole blood from 15 of the subjects (Welsch and Granoff, Clinical and Vaccine Immunology, 2007). Depending on the test strain, protective serum BA titers 1:4 were present in only 8 to 15 percent of these unvaccinated subjects as compared with 40 to 87 percent with the blood assay. Among serum BA-negative subjects, blood from 23 to 42 percent of the subjects gave a decrease of 2 log10 CFU/ml after one-hour incubation. Thus, while a serum BA titer 1:4 predicts protection against meningococcal disease, a titer <1:4 may be poorly predictive of susceptibility. The ability of whole blood from donors with serum bactericidal titers <1:4 to be bactericidal against N. meningitidis can be explained by the presence of nonbactericidal opsonic antibodies; or the presence of complement-mediated bactericidal antibodies below the threshold of detection of the serum bactericidal assay. Our data were consistent with both explanations since in some subjects with serum BA titers <1:4, bactericidal activity was present in 90% plasma (Figure 23, Donor 7), which could have resulted from the higher concentrations of antibody and/or complement present as compared with the assay conditions used for measuring BA in sera diluted 1:4. In other subjects (Figure 23, Donor 4), BA was observed in the whole blood assay but was greatly decreased or not observed at all in the 90% plasma bactericidal assay, which implied that white cells also were needed.

Figure 23. Bactericidal activity of whole blood or plasma against N. meningitidis group B, strain BZ232. Both donors has serum bactericidal titers <1:4. Solid lines and circles, bacteria incubated with whole blood, which showed effective killing. Dashed lines with triangles, bacteria incubated in 91 percent plasma that had been stored frozen to preserve internal complement activity. Plasma alone from donor 7 was sufficient for killing, but not from donor 4. (From Welsch and Granoff, Clin Vaccine Immunol 2007)

Meningococcal Vaccine Research Laboratory Program Description

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The possible role of opsonophagocytosis (OP) in protecting persons with negative serum BA titers remains controversial, in part because patients with deficiencies in terminal complement pathway proteins, whose sera support OP but not BA, have a greatly increased risk of acquiring meningococcal disease. To investigate the potential independent roles of serum BA and OP in conferring protection against group B meningococcal disease, we developed an OP bactericidal assay that used normal human serum depleted of C6 as an exogenous complement source, which in the absence of polymorphonuclear leukocytes (PMNs) did not support serum BA (Plested and Granoff, Clinical and Vaccine Immunology, 2008). We also assayed BA using C6sufficient complement, which in the absence of PMNs elicited bacteriolysis by formation of a membrane attack complex. We used these assays to measure BA and OP bactericidal activity of stored serum samples from 32 adults who had been immunized in a previous study with a detergent-treated outer membrane vesicle (OMV) vaccine given alone or combined with an investigational recombinant protein, genome-derived Neisserial antigen (GNA) 2132 (Welsch et al, Journal of Infectious Diseases, 2003). The test sera were heat-inactivated to remove intrinsic complement activity. Before immunization, 9 to19 percent of subjects in both vaccine groups had serum BA titers 1:4, which increased to 41 to 72 percent after immunization (P<0.01 against each of three test strains). The respective percentages of sera with OP titers were 3 to 16 before vaccination, which increased to 55 to 72 after three doses of vaccine (P<0.001 for each strain). For each of the three meningococcal strains tested there were examples of sera with BA titers <1:4 that were positive for OP bactericidal activity. Examples are shown in Figure 24 for a subject given the OMV vaccine alone (Panel A), and for a subject given the combination OMV/GNA2132 vaccine (Panel B). When the two vaccine groups were combined, the percentages of postimmunization BA-negative sera that were positive for OP bactericidal activity were 33 (3/9), 10 (1/10) and 37 (7/19) for strains H44/76, S3032 and NZ98/254, respectively. Collectively, the data supported independent roles for serum BA or OP bactericidal activity in protection against group B disease. Figure 24. Examples of two individual subjects with serum OP bactericidal activity in the presence or absence of complement-mediated BA. Percent survival (mean 95% CI) of different group B strains after 1 hour incubation with a 1:4 dilution of heat-inactivated test serum and C6-sufficient human serum as the complement source for assay BA (left, pre or post), or in an OP assay with a 1:5 dilution of the heat-inactivated test serum, human PMNs and C6-depleted human serum as the complement source (right, pre or post + PMNs). Data represent results of four replicate experiments for each test serum. Panel A, subject immunized with OMV vaccine. Panel B, subject immunized with the combination OMV/GNA2132 vaccine. From Plested and Granoff, Clin Vaccine Immuol 2008.