Assignment ofNeisseria meningitidisSerogroups A, C, W135, and Y Anticapsular Total Immunoglobulin G (IgG), IgG1, and IgG2 Concentrations to Reference Sera

Abstract

Meningococcal serogroup-specific immunoglobulin G (IgG), IgG1, and IgG2 concentrations were assigned to three reference sera, CDC 1992, 89-SF, and 96/562, for meningococcal serogroups A, C, Y, and W135 via the method of cross standardization. The sum of the serogroup-specific IgG1 and IgG2 concentrations determined for the four meningococcal serogroups showed good agreement with the serogroup-specific IgG either determined here or as previously represented. Following the assignment of meningococcal serogroup-specific IgG1 and IgG2 concentration to these reference sera, a meningococcal serogroup-specific IgG1 and IgG2 enzymelinked immunosorbent assay protocol was developed. The serogroup A and C specific subclass distribution of a panel of adult sera collected following vaccination with any combination of meningococcal serogroup C conjugate, bivalent, or tetravalent polysaccharide vaccines was determined. For the determination of serogroup W135 and Y specific subclass distribution, an adolescent panel 28 days following a single dose of either tetravalent polysaccharide or conjugate vaccine was used. The sum of the serogroup-specific IgG1 and IgG2 showed strong correlation with the serogroup-specific total IgG determined. The assignment here of IgG1 and IgG2 subclasses to these reference sera will allow more detailed evaluation of meningococcal conjugate and polysaccharide vaccines.

Development of a Fluorescent-Bead-Based Multiplex Immunoassay To Determine Immunoglobulin G Subclass Responses to Neisseria meningitidis Serogroup A and C Polysaccharides

“…The MenA and -C PS subclass MIA was performed as described previously (15) (11,12). Standard and serum samples were added to an equal volume of MenA-and -C-conjugated beads (4,000 beads/antigen/well) in a 96-well MV Multiscreen filter plate (Millipore, MA) and incubated for 20 min at room temperature (RT) on a plate shaker at 650 rpm in the dark.…”

“…MenA-and -C-specific IgG, IgG1, and IgG2 ELISA were performed as described previously (2,7,12), with the modification that different secondary conjugated antibodies were used: a goat anti-human IgG alkaline phosphatase-labeled antibody (Biosource, Camarillo, CA) and a rabbit antimouse Ig horseradish peroxidase-labeled antibody (DAKO, Glostrup, Denmark).…”

“…Conventional methods for detection of IgG and the IgG subclasses are based on enzyme-linked immunosorbent assays (ELISA) (2,7,12). ELISA is a reproducible and specific assay but time-consuming and often limited by the amount of available serum, particularly when multiple analytes need to be tested.…”

Induction of Protective Serum Meningococcal Bactericidal and Diphtheria-Neutralizing Antibodies and Mucosal Immunoglobulin A in Volunteers by Nasal Insufflations of theNeisseria meningitidisSerogroup C Polysaccharide-CRM197 Conjugate Vaccine Mixed with Chitosan

“…We observed a predominantly IgG2 response to MCP, with less IgG1 and almost no IgG3 or IgG4 whether the antigen was delivered nasally or parenterally. This pattern has been observed in studies of parenteral immunization with MCP (18,19). In contrast, CRM197-specific IgG subclasses were mixed, with equal levels of IgG1 and IgG2 and high levels of IgG4 in many subjects, probably indicating repeated boosting.…”

Development of a Multi-Plex Electrochemiluminescent Assay for the Detection of Serum Antibody Responses to Meningococcal Conjugate Vaccines

“…To best evaluate and optimize the assay, we used standard reference sera [15,17] as well as evaluating serum antibody responses to licensed vaccines in infant rhesus monkeys, a clinically relevant animal model for human infant vaccination. Both CDC 1992 and 89SF reference sera have been used to evaluate and standardize antibody based assays [22,23].…”