Flow cytometric assay for the measurement of serum opsonins to Neisseria meningitidis serogroup B, serotype 15

• Published in: Journal of immunological methods Vol. 116; no. 2; p. 235
• Main Authors: Sjursen, H, Bjerknes, R, Halstensen, A, Naess, A, Sørnes, S, Solberg, C O
• Format: Journal Article
• Language: English
• Published: Netherlands 17-01-1989
• Subjects: Blood Bactericidal Activity; Flow Cytometry; Humans; Meningitis, Meningococcal - immunology; Neisseria meningitidis - immunology; Opsonin Proteins - analysis; Phagocytes - physiology; Phagocytosis; Temperature; Time Factors
• ISSN: 0022-1759
• DOI: 10.1016/0022-1759(89)90209-3

A flow cytometric phagocytosis assay has been developed for the measurement of human serum opsonins to serogroup B meningococci. Live bacteria and bacteria inactivated by heat, formalin or ethanol were labelled with fluorescein-isothiocyanate (FITC). The bacteria were opsonized with sera from patients with group B meningococcal disease and sera from healthy controls, and phagocytosis determined by combined measurements of FITC-fluorescence and forward angle light scatter. Optimal sensitivity was obtained using viable bacteria, 5% serum, 20 bacteria per leukocyte capable of phagocytosis, 7.5 min opsonization time, 5 min phagocytosis time, 37 degrees C, and continuous agitation during opsonization and phagocytosis. The opsonic activity of sera from convalescent patients was markedly higher than that of sera from patients with acute illness. Only minor day-to-day and interindividual variations were observed. The flow cytometric phagocytosis technique is a rapid and reproducible method for the measurement of serum opsonins to meningococci.