The interaction in vitro between human polymorphonuclear leukocytes and Neisseria gonorrhoeae cultivated in the chick embryo.

Abstract

Gonococcal infections are a serious public health problem . Despite renewed public health efforts, the incidence of gonorrhea has continued to rise 10-15% annually since 1967 (1) . Examination of clinical specimens from infected patients often reveals gonococci closely associated with polymorphonuclear leukocytes . Despite this long-standing clinical observation, factors related to gonococci-human leukocyte interactions have remained undefined. This has been in large part due to the lack of an experimental animal and a suitable method for the laboratory cultivation of these organisms .Kellogg and co-workers described colonial variation of gonococci and based on direct inoculation of organisms into the male urethra correlated type 1 and 2 organisms with virulence and type 3 and 4 gonococci with avirulence . Nonselective transfer of type 1 and 2 gonococci on artificial medium resulted in reversion to type 3 and 4 organisms (2, 3) . Recent morphologic studies have suggested that colonial types 1 and 2 gonococci are more resistant to phagocytosis than type 3 and 4 organisms (4-6) .This report describes a method for the laboratory cultivation of gonococci which allows for quantitative in vitro phagocytosis studies of viable organisms. The findings indicate that log-phase virulent gonococci possess antiphagocytic surface factors and that avirulent organisms are ingested and rapidly killed by human polymorphonuclear leukocytes . Heat-labile serum factors are necessary for phagocytosis of log-phase organisms. In addition, log-phase virulent gonococci attach to the surface membrane but are not interiorized by human polymorphonuclear leukocytes in the presence or absence of fresh human serum.

Human deficiency of the eighth component of complement. The requirement of C8 for serum Neisseria gonorrhoeae bactericidal activity.

“…The importance of N. gonorrhoeae pili or other gonococcal cell surface antiphagocytic substances has yet to be clarified (34)(35)(36)(37)(38). Regardless, opsonization with phagocytosis may be of secondary importance compared to protective serum bactericidal mechanisms.…”

Resistance of Neisseria gonorrhoeae Grown in vivo to Ingestion and Digestion by Phagocytes of Human Blood

“…Pilated, small-colony gonococcal types resembling the virulent Kellogg types I and 2 (Kellogg et al, 1963(Kellogg et al, , 1968 seem to be less easily ingested by phagocytes than non-pilated, large-colony types resembling the avirulent Kellogg types 3 and 4 (Punsalang & Sawyer, 1973;Ofek, Beachey & Bisno, 1974;Dilworth, Hendley & Mandell, 1975;Gibbs & Roberts, 1975;)~ with the exception of the non-pilated large-colony type described by Swanson et al (1974) which is of unknown virulence for man. In some reports a clear distinction has been made between intracellular and cell-attached gonococci, e.g.…”

“…In some reports a clear distinction has been made between intracellular and cell-attached gonococci, e.g. by electron microscopy , by phase-contrast microscopy (Gibbs & Roberts, 1975) and by fluorescent antibody staining (Dilworth et al, 1975). Thus, resistance to ingestion appears to be correlated with the virulence for man of the Kellogg types, but whether or not pili are responsible for the resistance is still a matter of dispute (Swanson et al, 1974).…”

Penetration of Penicillin into Human Phagocytes Containing Neisseria gonorrhoeae: Intracellular Survival and Growth at Optimum Concentrations of Antibiotic

“…The intracellular viability of gonococci in vitro has been followed by incubating urethral pus, or human buffy coat phagocytes that had ingested gonococci (Watt, 1970;Ward et al, 1972;Gibbs & Roberts, 1975)~ with I pg penicillin/ml in order to kill extracellular gonococci. As viable gonococci disappeared in I to 3 h, gonococci seemingly could not survive within human phagocytes.…”