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Clinical and Diagnostic Laboratory Immunology, March 2001, p. 245-250, Vol. 8, No. 2
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.2.245-250.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Comparison of a Classical Phagocytosis Assay and a Flow Cytometry Assay for Assessment of the Phagocytic Capacity of Sera from Adults Vaccinated with a Pneumococcal Conjugate Vaccine

Wouter T. M. Jansen,1,* Merja Väkeväinen-Anttila,2 Helena Käyhty,2 Moon Nahm,3 N. Bakker,1 Jan Verhoef,1 Harm Snippe,1 and André F. M. Verheul1,dagger

Eijkman-Winkler Institute for Microbiology, Infectious Diseases, and Inflammation, Vaccines Section, University Medical Center, Utrecht, The Netherlands1; National Public Health Institute (KTL), Helsinki, Finland2; and University Rochester Medical Center, Rochester, New York3

Received 22 May 2000/Returned for modification 20 September 2000/Accepted 21 November 2000

Antibody- and complement-mediated phagocytosis is the main defense mechanism against Streptococcus pneumoniae. A standardized, easy to perform phagocytosis assay for pneumococci would be a great asset for the evaluation of the potential efficacy of (experimental) pneumococcal vaccines. Such an assay could replace the laborious phagocytosis assay of viable pneumococci (classical killing assay). Therefore, a newly developed phagocytosis assay based on flow cytometry (flow assay) was compared with the conventional killing assay and enzyme-linked immunosorbent assay (ELISA), using sera obtained from adults pre- and postvaccination with either a bivalent conjugate, a tetravalent conjugate, or the 23-valent polysaccharide vaccine. Highly significant correlations were observed between flow assay phagocytosis titers, killing assay phagocytosis titers, and ELISA antibody titers for serotype 6B and 23F as well. For serotype 19F, strong correlations were only observed between killing assay and ELISA titers. A potential drawback of the flow assay might be the low sensitivity compared with that of the killing assay. The choice of what assay to use, however, will depend on the objectives of the assay. When speed, easy performance, sample throughput, improved worker safety, absence of influence of antibiotics, and absence of false positives are the major criteria, the flow assay is the method of choice. When higher sensitivity is the major requirement, the classical killing assay should be used.

* Corresponding author. Mailing address: Eijkman-Winkler Institute for Microbiology, Infectious Diseases, and Inflammation, Vaccines Section, Utrecht Medical Center, Heidelberglaan 100, 3584 CX Utrecht. Phone: 31 30 2506534. Fax: 31 30 2541770. E-mail: W.T.M.Jansen{at}lab.azu.nl.

dagger Present address: Intervet International BV, Boxmeer, The Netherlands.

Clinical and Diagnostic Laboratory Immunology, March 2001, p. 245-250, Vol. 8, No. 2
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.2.245-250.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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