Pneumococcal Type 22F Polysaccharide Absorption Improves the Specificity of a Pneumococcal-Polysaccharide Enzyme-Linked Immunosorbent Assay

Abstract

The specificity of the immune response to the 23-valent pneumococcal-polysaccharide (PS) vaccine in healthy adults and to a pneumococcal conjugate vaccine in infants was examined by measuring immunoglobulin G (IgG) antibody titers by enzyme-linked immunosorbent assay (ELISA) and the opsonophagocytosis assay. ELISA measures total antipneumococcal IgG titers including the titers of functional and nonfunctional antibodies, while the opsonophagocytosis assay measures only functional-antibody titers. Twenty-four pairs of pre-and post-pneumococcal vaccination sera from adults were evaluated (ELISA) for levels of IgG antibodies against serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F. Twelve of the pairs were also examined (opsonophagocytosis assay) for their functional activities. The correlation coefficients between assay results for most types ranged from 0.75 to 0.90, but the correlation coefficient was only about 0.6 for serotypes 4 and 19F. The specificities of these antibodies were further examined by the use of competitive ELISA inhibition. A number of heterologous polysaccharides (types 11A, 12F, 15B, 22F, and 33A) were used as inhibitors. Most of the sera tested showed cross-reacting antibodies, in addition to those removed by pneumococcal C PS absorption. Our data suggest the presence of a common epitope that is found on most pneumococcal PS but that is not absorbed by purified C PS. Use of a heterologous pneumococcal PS (22F) to adsorb the antibodies to the common epitope increased the correlation between the IgG ELISA results and the opsonophagocytosis assay results. The correlation coefficient improve from 0.66 to 0.92 for type 4 and from 0.63 to 0.80 for type 19F. These common-epitope antibodies were largely absent in infants at 7 months of age, suggesting the carbohydrate nature of the epitope.

Schedules for Pneumococcal Vaccination of Preterm Infants: An RCT

“…After extraction from whole blood, sera were stored at -70°C before assaying for pneumococcal serotype-specific IgG concentrations for the PCV13 pneumococcal serotypes by using an enzyme-linked immunosorbent assay as previously described. 18 The lower limit of assay quantification was 0.15 μg/mL, and IgG concentrations ≥0.35 μg/mL were considered protective. 19 …”

Additive effect of pneumococcal vaccine and influenza vaccine on acute exacerbation in patients with chronic lung disease

“…Carl Frasch, was adsorbed to CWPS, but all other samples were adsorbed to CWPS (5 μg/ml) and 22F PPS (10 μg/ml) [25]. Serotype-specific IgG was determined for the four serotypes (6B, 14, 19F and 23F) that are the most prevalent among adult patients with pneumococcal infections in the US and Japan [5,26].…”

Relationship between IgG titers and opsonocytophagocytic activity of anti-pneumococcal antibodies after immunization with the 7-valent conjugate vaccine in allogeneic stem cell transplant

“…Serum pneumococcal IgG responses to the seven vaccine serotypes were determined by the WHO-standardized serotypespecific anti-pneumococcal capsular polysaccharide ELISA using pneumococcal C-polysaccharide and 22F to adsorb non-specific antibodies. 10,11 Pneumococcal IgG ELISA titers were expressed as mg/ml using the standard reference serum 89-SF.…”