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Clinical and Diagnostic Laboratory Immunology, March 2001, p. 266-272, Vol. 8, No. 2
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.2.266-272.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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

Nelydia F. Concepcion and Carl E. Frasch*

Laboratory of Bacterial Polysaccharides, Center for Biologics Evaluation and Research, Bethesda, Maryland

Received 24 July 2000/Returned for modification 9 October 2000/Accepted 21 November 2000

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.

* Corresponding author. Mailing address: Division of Bacterial Products, CBER, 1401 Rockville Pike, HFM-428, Rockville, MD 20852. Phone: (301) 496-1920. Fax: (301) 402-2776. E-mail: frasch{at}cber.fda.gov.

Clinical and Diagnostic Laboratory Immunology, March 2001, p. 266-272, Vol. 8, No. 2
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.2.266-272.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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Copyright © 2001 by the American Society for Microbiology. All rights reserved.