title: Characterization of specific immunoglobulin G (IgG) and its subclasses (IgG1 and IgG2) against the 23-valent pneumococcal vaccine in a healthy adult population: proposal for response criteria.
abstract: The aim of the study was to standardize an enzyme-linked immunosorbent assay (ELISA) method for the quantification of immunoglobulin G (IgG) and its subclasses (IgG1 and IgG2) against the 23-valent pneumococcal vaccine and to establish the criteria for a normal response to the vaccine. Forty healthy individuals (20 women and 20 men; mean age, 29 years) were studied. All were vaccinated with the 23-valent pneumococcal vaccine; blood samples were drawn just prior to and 3 weeks after immunization. Quantification of specific IgG and its subclasses was performed by an ELISA with the vaccine as the antigen. The linearity of the ELISA method was demonstrated by the similar slopes of the linear regression lines generated from the titration of sera with different antibody concentrations. The specificity of the antibodies against the vaccine was demonstrated by (i) an absorption test with pneumococcal vaccine, (ii) a cross-reactivity experiment with Haemophilus influenzae type b polysaccharide, and (iii) affinity chromatography with protein A-Sepharose. Response to the vaccine was defined by using the lower level of the 90% probability interval (one-tailed) for postimmunization-specific IgG, IgG1, and IgG2. By using this cutoff, responders were considered to be those with an absolute increase in antibody titers higher than 395 arbitrary units/ml for IgG, 0.350 A450 units for IgG1, and 0.314 A450 units for IgG2. Overall, 20 (50%) subjects had IgG, IgG1, and IgG2 responses, 9 (22.5%) had IgG and IgG2 responses, 4 (10%) had IgG1 responses, 3 (7.5%) had IgG and IgG1 responses, and 4 (10%) were nonresponders. Ninety percent of our population responded to the 23-valent pneumococcal vaccine. Up to 10% of healthy individuals may respond to an IgG subclass without significant increases in total IgG titers. The ELISA method that is described may be useful for evaluating the specific antibody response against polysaccharides.
authors.affiliation: Department of Biochemistry (Immunology Unit), Hospital General Vall d'Hebron, Barcelona, Spain.
authors.forename: M J
authors.initials: MJ
authors.lastname: Rodrigo
authors.forename: M
authors.initials: M
authors.lastname: Miravitlles
authors.forename: M J
authors.initials: MJ
authors.lastname: Cruz
authors.forename: J
authors.initials: J
authors.lastname: de Gracia
authors.forename: M
authors.initials: M
authors.lastname: Vendrell
authors.forename: C
authors.initials: C
authors.lastname: Pascual
authors.forename: F
authors.initials: F
authors.lastname: Morell
mesh_terms: D000328:Adult; D000907:Antibodies, Bacterial / Q000097:blood*; D000918:Antibody Specificity; D001428:Bacterial Vaccines / Q000276:immunology*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards*; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood*; D007163:Immunosorbent Techniques; D008297:Male; D008875:Middle Aged; D022242:Pneumococcal Vaccines; D012015:Reference Standards; D013296:Streptococcus pneumoniae / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D001428:Bacterial Vaccines; D007074:Immunoglobulin G; D022242:Pneumococcal Vaccines