title: A modified surface killing assay (MSKA) as a functional in vitro assay for identifying protective antibodies against pneumococcal surface protein A (PspA).
abstract: Streptococcus pneumoniae causes otitis media, meningitis and pneumonia in patients worldwide; predominantly affecting young children, the elderly, and the immune compromised. Current vaccines against invasive pneumococcal disease are based on the polysaccharide capsules of the most clinically relevant serotypes. Due to serotype replacement, non-vaccine serotypes of S. pneumoniae have become more clinically relevant and as a result pneumococcal vaccines are becoming increasingly complex. These events emphasize the need to evaluate the potential for pneumococcal cross-reactive proteins to contribute to future vaccines. Antibody elicited by the immunization of humans with pneumococcal surface protein A (PspA) can passively protect mice from infection. However, robust in vitro functional assays for antibody to PspA are not available to predict the protective capacity of immune serum. For polysaccharide based vaccines, a standardized opsonophagocytosis killing assay (OPKA) is used. Antibody to PspA, however, does not work well in the standard OPKA. The present studies take advantage of past observations that phagocytosis is more efficient on tissue surfaces than in solution. In a modified surface killing assay (MSKA), monoclonal antibody to PspA, in the presence of complement, opsonized pneumococci for killing by phagocytes on an agar surface. Five monoclonal antibodies to PspA were tested; three demonstrated increased amounts of killing compared to the diluent control and protected mice by passive protection against type 3 pneumococci. The two antibodies that were not functional in the MSKA also failed to protect mice. Thus, an MSKA might be useful as a functional assay for immunity to PspA.
authors.affiliation: Department of Microbiology, University of Alabama at Birmingham, Birmingham, AL 35294, United States.
authors.forename: Kristopher R
authors.initials: KR
authors.lastname: Genschmer
authors.affiliation: Department of Genetics, University of Alabama at Birmingham, Birmingham, AL 35294, United States.
authors.forename: Mary Ann
authors.initials: MA
authors.lastname: Accavitti-Loper
authors.affiliation: Department of Microbiology, University of Alabama at Birmingham, Birmingham, AL 35294, United States; Department of Genetics, University of Alabama at Birmingham, Birmingham, AL 35294, United States; Department of Pediatrics, University of Alabama at Birmingham, Birmingham, AL 35294, United States. Electronic address: dbriles@uab.edu.
authors.forename: David E
authors.initials: DE
authors.lastname: Briles
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000097:blood / Q000276:immunology*; D001426:Bacterial Proteins / Q000276:immunology*; D006801:Humans; D007116:Immunization, Passive; D051379:Mice; D010587:Phagocytosis / Q000276:immunology; D011008:Pneumococcal Infections / Q000276:immunology* / Q000517:prevention & control; D022242:Pneumococcal Vaccines / Q000276:immunology; D013296:Streptococcus pneumoniae / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D001426:Bacterial Proteins; D022242:Pneumococcal Vaccines; C051592:pneumococcal surface protein A
keywords: Functional assay; Pneumococcal surface protein A; Polymorphonuclear leukocytes; PspA; Streptococcus pneumoniae; Surface killing assay