Monocyte phagocytosis of opsonized Neisseria meningitidis serogroup B.
The chemiluminescence (CL) was examined when peripheral blood monocytes were incubated with opsonized Neisseria meningitidis, serogroup B, serotype 15:P1.16 or serotype 2a:P1.2. The monocytes were separated from a mononuclear cell suspension by an immunomagnetic negative selection technique using magnetic polystyrene microspheres coated with monoclonal antibodies specific for T and B lymphocytes. More than 90% of the lymphocytes were removed, yielding a suspension containing 93% monocytes. Optimal sensitivity for phagocytosis was obtained using 1% serum (10 microliters), 72 bacteria per monocyte cell, and 7.5 min opsonization and incubation time during continuous agitation at 37 degrees C. The CL was amplified by lucigenin. Preliminary experiments suggest that convalescent sera from patients with group B meningococcal disease induced increased CL responses compared to acute sera. Sera from volunteers immunized with an outer membrane complex vaccine from serogroup B, serotype 15:P1.16 or 2a:P1.2 meningococci also induced increased CL activity compared to preimmune sera. No such response was shown when a group B capsular polysaccharide vaccine was given. This response pattern was also demonstrated by a flow cytometric phagocytosis technique (FCM). Internalization of meningococci by monocytes was demonstrated by a FCM quenching technique and by transmission electron microscopy. CL and FCM represent rapid and reproducible methods for the measurement of opsonophagocytosis of meningococci by monocytes and may be performed with minute amounts of sera.
Sjursen
H
H
Medical Department B, Tromsø Regional Hospital, Norway.
Lehmann
V
V
Naess
A
A
Hervig
T
T
Flø
R W
RW
Maehle
B
B
Halstensen
A I
AI
Frøholm
L O
LO
D000328:Adult; D002469:Cell Separation / Q000379:methods; D005434:Flow Cytometry; D006801:Humans; D008163:Luminescent Measurements; D008854:Microscopy, Electron; D008863:Microspheres; D009000:Monocytes / Q000276:immunology* / Q000648:ultrastructure; D009345:Neisseria meningitidis / Q000276:immunology*; D010587:Phagocytosis