Phase Transition of Gonococci in Mammalian Cell Cultures

Tyeryar, Franklin J.; Quan, Alice L.; René, Anthony A.; Weiss, Emilio

doi:10.1128/iai.10.6.1401-1411.1974

Cited by 11 publications

(6 citation statements)

References 21 publications

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“…4). It is difficult to determine on the basis of these micrographs whether the gonococci situated in the cytoplasm of the HeLa cells are enclosed within membrane-limited vacuoles as observed by others (26,34,35). Organisms adhering to the tissue cell surface have portions of their outer membrane that are closely integrated with the cytoplasmic membranes of the tissue cell (Fig.…”

Section: Resultsmentioning

confidence: 93%

“…Although serum bactericidal factors have been demonstrated in vitro, they may not always operate in vivo. It would seem that gonococcal pathogenicity may involve other factors such as survival of gonococci within human polymorphonuclear leukocytes (4,18,24,32,38) or within normal eukaryotic cells (21,25,30,31,34,35). Furthermore, electron microscopy of infected tissues have provided strong evidence of intracellular localization of gonococci in urethral exudate (8,20) as well as in cultures of infected HeLa cells (34), human diploid cells (25), and monkey kidney cells (35).…”

mentioning

confidence: 99%

“…It would seem that gonococcal pathogenicity may involve other factors such as survival of gonococci within human polymorphonuclear leukocytes (4,18,24,32,38) or within normal eukaryotic cells (21,25,30,31,34,35). Furthermore, electron microscopy of infected tissues have provided strong evidence of intracellular localization of gonococci in urethral exudate (8,20) as well as in cultures of infected HeLa cells (34), human diploid cells (25), and monkey kidney cells (35). Correlation of adsorption to tissue cells and type variation of N. gonorrhoeae (15) has shown that virulent type 1 demonstrates enhanced attachment to various types of host cells (13,26,28,33,37) when compared with avirulent type 4. It has been observed that attachment and passage into underlying tissues through mucosal cells is important in virulent Shigella infections (2).…”

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confidence: 99%

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In vitro interaction of Neisseria gonorrhoeae type 1 and type 4 with tissue culture cells

Brodeur¹,

Johnson²,

Johnson³

et al. 1977

Infect Immun

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As a basis for studies of gonococcal pathogenicity, tissue culture cells were infected with type 1 or type 4 Neisseria gonorrhoeae to determine intracellular viability. A simple and objective means of measurement was devised, based on the uptake of tritiated protein and deoxyribonucleic acid precursors by cycloheximide-inhibited cells infected with gonococci. Cycloheximide was found to inhibit protein synthesis by over 97% in tissue culture cells at a concentration of 100 jig/ml. In contrast, N. gonorrhoeae was found to be highly resistant to this antibiotic, and protein synthesis was unaffected by concentrations up to 1,000 ,g/ml. Extracellular gonococci were eliminated by treatment with high concentrations of penicillin during cycloheximide inhibition and prior to the addition of radioisotope. Levels of protein and deoxyribonucleic acid synthesis by N. gonorrhoeae in the cycloheximide-treated cells were significantly higher in T1-infected cells (RE2, HeLa, or HEp-2) than in the corresponding T4-infected cells. No differences were observed in tissue cell susceptibility to gonococcal infection. Intracytoplasmic localization of N. gonorrhoeae was confirmed by electron microscopy.

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Section: Resultsmentioning

confidence: 93%

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confidence: 99%

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confidence: 99%

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In vitro interaction of Neisseria gonorrhoeae type 1 and type 4 with tissue culture cells

Brodeur¹,

Johnson²,

Johnson³

et al. 1977

Infect Immun

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“…A number of investigators have focused upon gonococcal interaction with tissue culture monolayers. Depending on the tissue culture cell type, gonococci were found only to adhere (17) or to adhere or enter other cells (20,22). Bessen and Gotschlich (1) found that Pll variants of N. gonorrhoeae R10 differed in the ability to adhere to HeLa cells.…”

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confidence: 99%

Model for invasion of human tissue culture cells by Neisseria gonorrhoeae

1988

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A tissue culture model has been developed for studying the ability of Neisseria gonorrhoeae to invade eucaryotic cells. The cell line HecIB, a human adenocarcinoma endometrial cell line, was found to support gonococcal invasion. The bactericidal antibiotic gentamicin was used to kill those bacteria that had not entered the HecIB cells, allowing us to quantitate internalized bacteria. Kinetic studies showed an increase in the titer of gentamicin-protected gonococci at 4 h postinfection followed by a decrease; a second increase occurred after 6 h. The state of piliation did not affect the degree of invasion when the bacteria were spun down onto the monolayer. Gonococcal invasion was inhibited when the HecIB cells were preincubated with cytochalasin D before bacterial infection. N. lactamica was used as a negative control. No internalized N. lactamica cells were observed by electron microscopy. Electron microscopy documented the intracellular location of the gonococci in HecIB cells and the eventual destruction of the invaded HecIB cells. After 24 h, clusters of gonococci encased in a matrix of cell debris were observed.

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“…Opacity-associated proteins and pili each provide for increased gonococcal adherence to a variety of host cells in vitro (10,16,23,27). Electron micrographs of gonococci cultivated with monolayers of tissue culture cells show organisms enclosed in membrane-bound vacuoles within the cytoplasm (1,20,32,33). Tissue culture cells provide a model for some of the events which are likely to occur during infection in vivo.…”

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Interactions of gonococci with HeLa cells: attachment, detachment, replication, penetration, and the role of protein II

Bessen¹,

Gotschlich²

1986

Infect Immun

102

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Colony variants of Neisseria gonorrhoeae differ in their interactions with eucaryotic cells. When gonococci were cultivated with HeLa cell monolayers, the opacity phenotype (Op) became increasingly dominant in the subpopulation of organisms which adhered to the HeLa cells. Once bound, Op organisms displayed very low levels of detachment. Adherent Op gonococci exhibited generation times up to threefold greater than cultures containing gonococci in the absence of HeLa cells. In addition, the progeny of adherent Op organisms remained bound to the HeLa cell monolayer. Both piliated (P+) and transparent (Tr) colony types attached to HeLa cells, but their progeny were retained less efficiently. Gonococci bound to HeLa cells were subjected to the bactericidal action of fresh rat serum and approximately 0.5 to 2.5% survived, irrespective of their opacity or piliation phenotype. Incubation with gentamicin resulted in a 10- to 50-fold further reduction in viability. Pretreatment of HeLa cell monolayers with the microfilament-disrupting agent cytochalasin b diminished gonococcal survival in either serum or gentamicin by up to eightfold. In contrast, cytochalasin b treatment did not decrease survival of the commensal organism N. sicca. The data suggest that very few gonococci are completely interiorized and a small proportion of adherent gonococci are partially protected from the soluble-phase environment by HeLa cells.

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Phase Transition of Gonococci in Mammalian Cell Cultures

Abstract: Neisseria gonorrhoeae was cultivated in mammalian cell cultures in an effort to determine if this environment will elicit a T4 Ti transition. Of four avirulent (T4) isolates tested, only one, H4, yielded Ti colonies. This change was

Cited by 11 publications

References 21 publications

In vitro interaction of Neisseria gonorrhoeae type 1 and type 4 with tissue culture cells

In vitro interaction of Neisseria gonorrhoeae type 1 and type 4 with tissue culture cells

Model for invasion of human tissue culture cells by Neisseria gonorrhoeae

Interactions of gonococci with HeLa cells: attachment, detachment, replication, penetration, and the role of protein II

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