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. 1995 May;63(5):1754–1761. doi: 10.1128/iai.63.5.1754-1761.1995

Use of tissue culture and animal models to identify virulence-associated traits of Haemophilus ducreyi.

M J Alfa 1, M K Stevens 1, P DeGagne 1, J Klesney-Tait 1, J D Radolf 1, E J Hansen 1
PMCID: PMC173220  PMID: 7729882

Abstract

To identify virulence-associated properties of Haemophilus ducreyi, 34 strains of this sexually transmitted pathogen were evaluated for in vitro phenotypic characteristics of potential relevance to chancroid pathogenesis and for their ability to produce lesions in the temperature-dependent animal model for chancroid. Of the 34 strains tested, all but three produced a cytopathic effect on human foreskin fibroblasts (HFF) and all but six strains formed large microcolonies on HFF monolayers. A subset of 12 selected strains underwent more extensive analyses and, when evaluated for both their cytadherence kinetics and growth in the presence of HFF monolayers, it was found that several of these strains had a very limited ability to attach to HFF cells. When the same 12 strains were tested in the temperature-dependent rabbit model, only the seven strains which were positive in all of these in vitro-based tests readily produced lesions. In contrast, the five strains that were noted to be deficient in one or more of the phenotypic characteristics scored in the in vitro systems did not produce lesions. This association between the traits measured in vitro and the ability to produce dermal lesions was significant (P = 0.0012). These results suggest that in vitro behavior may be used to predict the virulence potential of H. ducreyi strains. Moreover, the phenotypic characteristics described in this study are appropriate focal points for efforts to determine the molecular basis of the virulence of this pathogen.

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Selected References

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  1. Abeck D., Johnson A. P., Hartinger A., Kollmann M., Korting H. C., Ballard R. C., Braun-Falco O. Absence of extracellular enzyme activity and cytotoxicity in cell-free culture filtrates of Haemophilus ducreyi. Microb Pathog. 1991 May;10(5):405–410. doi: 10.1016/0882-4010(91)90085-o. [DOI] [PubMed] [Google Scholar]
  2. Abeck D., Johnson A. P., Mensing H. Binding of Haemophilus ducreyi to extracellular matrix proteins. Microb Pathog. 1992 Jul;13(1):81–84. doi: 10.1016/0882-4010(92)90034-l. [DOI] [PubMed] [Google Scholar]
  3. Albritton W. L. Biology of Haemophilus ducreyi. Microbiol Rev. 1989 Dec;53(4):377–389. doi: 10.1128/mr.53.4.377-389.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Alfa M. J. Cytopathic effect of Haemophilus ducreyi for human foreskin cell culture. J Med Microbiol. 1992 Jul;37(1):43–50. doi: 10.1099/00222615-37-1-43. [DOI] [PubMed] [Google Scholar]
  5. Alfa M. J., Degagne P., Hollyer T. Haemophilus ducreyi adheres to but does not invade cultured human foreskin cells. Infect Immun. 1993 May;61(5):1735–1742. doi: 10.1128/iai.61.5.1735-1742.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Alfa M. J., Yang C. L., Slaney L. A., Kwok A. Y., Ronald A. R., Jay F. T. Identification of highly conserved and species-specific polypeptides of Haemophilus ducreyi. J Med Microbiol. 1992 Dec;37(6):413–419. doi: 10.1099/00222615-37-6-413. [DOI] [PubMed] [Google Scholar]
  7. Cameron D. W., Simonsen J. N., D'Costa L. J., Ronald A. R., Maitha G. M., Gakinya M. N., Cheang M., Ndinya-Achola J. O., Piot P., Brunham R. C. Female to male transmission of human immunodeficiency virus type 1: risk factors for seroconversion in men. Lancet. 1989 Aug 19;2(8660):403–407. doi: 10.1016/s0140-6736(89)90589-8. [DOI] [PubMed] [Google Scholar]
  8. Campagnari A. A., Wild L. M., Griffiths G. E., Karalus R. J., Wirth M. A., Spinola S. M. Role of lipooligosaccharides in experimental dermal lesions caused by Haemophilus ducreyi. Infect Immun. 1991 Aug;59(8):2601–2608. doi: 10.1128/iai.59.8.2601-2608.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Finlay B. B., Falkow S. Common themes in microbial pathogenicity. Microbiol Rev. 1989 Jun;53(2):210–230. doi: 10.1128/mr.53.2.210-230.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Greenblatt R. M., Lukehart S. A., Plummer F. A., Quinn T. C., Critchlow C. W., Ashley R. L., D'Costa L. J., Ndinya-Achola J. O., Corey L., Ronald A. R. Genital ulceration as a risk factor for human immunodeficiency virus infection. AIDS. 1988 Feb;2(1):47–50. doi: 10.1097/00002030-198802000-00008. [DOI] [PubMed] [Google Scholar]
  11. Hammond G. W., Lian C. J., Wilt J. C., Ronald A. R. Antimicrobial susceptibility of Haemophilus ducreyi. Antimicrob Agents Chemother. 1978 Apr;13(4):608–612. doi: 10.1128/aac.13.4.608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Handsfield H. H., Totten P. A., Fennel C. L., Falkow S., Holmes K. K. Molecular epidemiology of Haemophilus ducreyi infections. Ann Intern Med. 1981 Sep;95(3):315–318. doi: 10.7326/0003-4819-95-3-315. [DOI] [PubMed] [Google Scholar]
  13. Hansen E. J., Latimer J. L., Thomas S. E., Helminen M., Albritton W. L., Radolf J. D. Use of electroporation to construct isogenic mutants of Haemophilus ducreyi. J Bacteriol. 1992 Aug;174(16):5442–5449. doi: 10.1128/jb.174.16.5442-5449.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hansen E. J., Lumbley S. R., Richardson J. A., Purcell B. K., Stevens M. K., Cope L. D., Datte J., Radolf J. D. Induction of protective immunity to Haemophilus ducreyi in the temperature-dependent rabbit model of experimental chancroid. J Immunol. 1994 Jan 1;152(1):184–192. [PubMed] [Google Scholar]
  15. Jessamine P. G., Plummer F. A., Ndinya Achola J. O., Wainberg M. A., Wamola I., D'Costa L. J., Cameron D. W., Simonsen J. N., Plourde P., Ronald A. R. Human immunodeficiency virus, genital ulcers and the male foreskin: synergism in HIV-1 transmission. Scand J Infect Dis Suppl. 1990;69:181–186. [PubMed] [Google Scholar]
  16. Lagergård T. The role of Haemophilus ducreyi bacteria, cytotoxin, endotoxin and antibodies in animal models for study of chancroid. Microb Pathog. 1992 Sep;13(3):203–217. doi: 10.1016/0882-4010(92)90021-f. [DOI] [PubMed] [Google Scholar]
  17. Lammel C. J., Dekker N. P., Palefsky J., Brooks G. F. In vitro model of Haemophilus ducreyi adherence to and entry into eukaryotic cells of genital origin. J Infect Dis. 1993 Mar;167(3):642–650. doi: 10.1093/infdis/167.3.642. [DOI] [PubMed] [Google Scholar]
  18. Melaugh W., Phillips N. J., Campagnari A. A., Karalus R., Gibson B. W. Partial characterization of the major lipooligosaccharide from a strain of Haemophilus ducreyi, the causative agent of chancroid, a genital ulcer disease. J Biol Chem. 1992 Jul 5;267(19):13434–13439. [PubMed] [Google Scholar]
  19. Morse S. A. Chancroid and Haemophilus ducreyi. Clin Microbiol Rev. 1989 Apr;2(2):137–157. doi: 10.1128/cmr.2.2.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Odumeru J. A., Wiseman G. M., Ronald A. R. Relationship between lipopolysaccharide composition and virulence of Haemophilus ducreyi. J Med Microbiol. 1987 Mar;23(2):155–162. doi: 10.1099/00222615-23-2-155. [DOI] [PubMed] [Google Scholar]
  21. Odumeru J. A., Wiseman G. M., Ronald A. R. Role of lipopolysaccharide and complement in susceptibility of Haemophilus ducreyi to human serum. Infect Immun. 1985 Nov;50(2):495–499. doi: 10.1128/iai.50.2.495-499.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Odumeru J. A., Wiseman G. M., Ronald A. R. Virulence factors of Haemophilus ducreyi. Infect Immun. 1984 Feb;43(2):607–611. doi: 10.1128/iai.43.2.607-611.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Plummer F. A., Simonsen J. N., Cameron D. W., Ndinya-Achola J. O., Kreiss J. K., Gakinya M. N., Waiyaki P., Cheang M., Piot P., Ronald A. R. Cofactors in male-female sexual transmission of human immunodeficiency virus type 1. J Infect Dis. 1991 Feb;163(2):233–239. doi: 10.1093/infdis/163.2.233. [DOI] [PubMed] [Google Scholar]
  24. Purcell B. K., Richardson J. A., Radolf J. D., Hansen E. J. A temperature-dependent rabbit model for production of dermal lesions by Haemophilus ducreyi. J Infect Dis. 1991 Aug;164(2):359–367. doi: 10.1093/infdis/164.2.359. [DOI] [PubMed] [Google Scholar]
  25. Purvén M., Lagergård T. Haemophilus ducreyi, a cytotoxin-producing bacterium. Infect Immun. 1992 Mar;60(3):1156–1162. doi: 10.1128/iai.60.3.1156-1162.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Simonsen J. N., Cameron D. W., Gakinya M. N., Ndinya-Achola J. O., D'Costa L. J., Karasira P., Cheang M., Ronald A. R., Piot P., Plummer F. A. Human immunodeficiency virus infection among men with sexually transmitted diseases. Experience from a center in Africa. N Engl J Med. 1988 Aug 4;319(5):274–278. doi: 10.1056/NEJM198808043190504. [DOI] [PubMed] [Google Scholar]
  27. Spinola S. M., Castellazzo A., Shero M., Apicella M. A. Characterization of pili expressed by Haemophilus ducreyi. Microb Pathog. 1990 Dec;9(6):417–426. doi: 10.1016/0882-4010(90)90060-4. [DOI] [PubMed] [Google Scholar]
  28. Spinola S. M., Griffiths G. E., Bogdan J., Menegus M. A. Characterization of an 18,000-molecular-weight outer membrane protein of Haemophilus ducreyi that contains a conserved surface-exposed epitope. Infect Immun. 1992 Feb;60(2):385–391. doi: 10.1128/iai.60.2.385-391.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Spinola S. M., Griffiths G. E., Shanks K. L., Blake M. S. The major outer membrane protein of Haemophilus ducreyi is a member of the OmpA family of proteins. Infect Immun. 1993 Apr;61(4):1346–1351. doi: 10.1128/iai.61.4.1346-1351.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Spinola S. M., Wild L. M., Apicella M. A., Gaspari A. A., Campagnari A. A. Experimental human infection with Haemophilus ducreyi. J Infect Dis. 1994 May;169(5):1146–1150. doi: 10.1093/infdis/169.5.1146. [DOI] [PubMed] [Google Scholar]
  31. Sturm A. W., Zanen H. C. Characteristics of Haemophilus ducreyi in culture. J Clin Microbiol. 1984 May;19(5):672–674. doi: 10.1128/jcm.19.5.672-674.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Telzak E. E., Chiasson M. A., Bevier P. J., Stoneburner R. L., Castro K. G., Jaffe H. W. HIV-1 seroconversion in patients with and without genital ulcer disease. A prospective study. Ann Intern Med. 1993 Dec 15;119(12):1181–1186. doi: 10.7326/0003-4819-119-12-199312150-00005. [DOI] [PubMed] [Google Scholar]
  33. Totten P. A., Morton W. R., Knitter G. H., Clark A. M., Kiviat N. B., Stamm W. E. A primate model for chancroid. J Infect Dis. 1994 Jun;169(6):1284–1290. doi: 10.1093/infdis/169.6.1284. [DOI] [PubMed] [Google Scholar]
  34. Trees D. L., Arko R. J., Morse S. A. Mouse subcutaneous chamber model for in vivo growth of Haemophilus ducreyi. Microb Pathog. 1991 Nov;11(5):387–390. doi: 10.1016/0882-4010(91)90025-6. [DOI] [PubMed] [Google Scholar]
  35. Tuffrey M., Alexander F., Ballard R. C., Taylor-Robinson D. Characterization of skin lesions in mice following intradermal inoculation of Haemophilus ducreyi. J Exp Pathol (Oxford) 1990 Apr;71(2):233–244. [PMC free article] [PubMed] [Google Scholar]

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