Abstract
H. ducreyi is the causative agent of chancroid, a genital ulcer disease most prevalent in developing countries. Chancroid enhances the heterosexual transmission of human immunodeficiency virus and is identified in focal outbreaks in the United States, but little is known about its pathogenesis. We studied the hemolysin produced by H. ducreyi because this molecule might be an important virulence factor in the pathogenesis of chancroid. Ten strains of H. ducreyi were tested on newly devised blood agar plates and were found to have hemolytic activity. We examined the hemolytic activity of H. ducreyi 35000 further and found that it was heat labile, cell associated, greatest at pH 7.0, and produced in logarithmic- but not stationary-phase cultures. Using transposons Tn916 and Tn1545-delta 3, we have isolated three classes of transposon mutants of strain 35000: those with no detectable hemolytic activity, those with reduced hemolytic activity, and those with enhanced hemolytic activity. Transposon insertions in the nonhemolytic mutants were located in a DNA sequence which hybridized to the Proteus mirabilis hemolysin gene. Analysis of clones containing overlapping sections of this region served to further localize the H. ducreyi hemolysin gene and allow its expression in Escherichia coli and complementation of the nonhemolytic defect in an H. ducreyi mutant. These experiments indicate that H. ducreyi 35000 produces a hemolysin that is related to the calcium-independent hemolysin produced by P. mirabilis. Further experiments are needed to define the similarity of the H. ducreyi hemolysin to other calcium-independent hemolysins and to determine its role in the pathogenesis of chancroid.
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- 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]
- 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]
- Barenkamp S. J., St Geme J. W., 3rd Genes encoding high-molecular-weight adhesion proteins of nontypeable Haemophilus influenzae are part of gene clusters. Infect Immun. 1994 Aug;62(8):3320–3328. doi: 10.1128/iai.62.8.3320-3328.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Gawron-Burke C., Clewell D. B. Regeneration of insertionally inactivated streptococcal DNA fragments after excision of transposon Tn916 in Escherichia coli: strategy for targeting and cloning of genes from gram-positive bacteria. J Bacteriol. 1984 Jul;159(1):214–221. doi: 10.1128/jb.159.1.214-221.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grimminger F., Sibelius U., Bhakdi S., Suttorp N., Seeger W. Escherichia coli hemolysin is a potent inductor of phosphoinositide hydrolysis and related metabolic responses in human neutrophils. J Clin Invest. 1991 Nov;88(5):1531–1539. doi: 10.1172/JCI115463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Hughes H. P., Campos M., McDougall L., Beskorwayne T. K., Potter A. A., Babiuk L. A. Regulation of major histocompatibility complex class II expression by Pasteurella haemolytica leukotoxin. Infect Immun. 1994 May;62(5):1609–1615. doi: 10.1128/iai.62.5.1609-1615.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kauc L., Goodgal S. H. Introduction of transposon Tn916 DNA into Haemophilus influenzae and Haemophilus parainfluenzae. J Bacteriol. 1989 Dec;171(12):6625–6628. doi: 10.1128/jb.171.12.6625-6628.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- König W., Faltin Y., Scheffer J., Schöffler H., Braun V. Role of cell-bound hemolysin as a pathogenicity factor for Serratia infections. Infect Immun. 1987 Nov;55(11):2554–2561. doi: 10.1128/iai.55.11.2554-2561.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lally E. T., Golub E. E., Kieba I. R., Taichman N. S., Decker S., Berthold P., Gibson C. W., Demuth D. R., Rosenbloom J. Structure and function of the B and D genes of the Actinobacillus actinomycetemcomitans leukotoxin complex. Microb Pathog. 1991 Aug;11(2):111–121. doi: 10.1016/0882-4010(91)90004-t. [DOI] [PubMed] [Google Scholar]
- Lally E. T., Golub E. E., Kieba I. R., Taichman N. S., Rosenbloom J., Rosenbloom J. C., Gibson C. W., Demuth D. R. Analysis of the Actinobacillus actinomycetemcomitans leukotoxin gene. Delineation of unique features and comparison to homologous toxins. J Biol Chem. 1989 Sep 15;264(26):15451–15456. [PubMed] [Google Scholar]
- 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]
- Moseley S. L., Falkow S. Nucleotide sequence homology between the heat-labile enterotoxin gene of Escherichia coli and Vibrio cholerae deoxyribonucleic acid. J Bacteriol. 1980 Oct;144(1):444–446. doi: 10.1128/jb.144.1.444-446.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nassif X., Puaoi D., So M. Transposition of Tn1545-delta 3 in the pathogenic Neisseriae: a genetic tool for mutagenesis. J Bacteriol. 1991 Apr;173(7):2147–2154. doi: 10.1128/jb.173.7.2147-2154.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Oberhofer T. R., Back A. E. Isolation and cultivation of Haemophilus ducreyi. J Clin Microbiol. 1982 Apr;15(4):625–629. doi: 10.1128/jcm.15.4.625-629.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Ondraczek R., Hobbie S., Braun V. In vitro activation of the Serratia marcescens hemolysin through modification and complementation. J Bacteriol. 1992 Aug;174(15):5086–5094. doi: 10.1128/jb.174.15.5086-5094.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Palmer K. L., Grass S., Munson R. S., Jr Identification of a hemolytic activity elaborated by Haemophilus ducreyi. Infect Immun. 1994 Jul;62(7):3041–3043. doi: 10.1128/iai.62.7.3041-3043.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Portnoy D. A., Chakraborty T., Goebel W., Cossart P. Molecular determinants of Listeria monocytogenes pathogenesis. Infect Immun. 1992 Apr;60(4):1263–1267. doi: 10.1128/iai.60.4.1263-1267.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Prince A. S., Barlam T. Isolation of a DNA fragment containing replication functions from IncP2 megaplasmid pMG2. J Bacteriol. 1985 Feb;161(2):792–794. doi: 10.1128/jb.161.2.792-794.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Rennie R. P., Arbuthnott J. P. Partial characterisation of Escherichia coli haemolysin. J Med Microbiol. 1974 May;7(2):179–188. doi: 10.1099/00222615-7-2-179. [DOI] [PubMed] [Google Scholar]
- Roberts M., Stull T. L., Smith A. L. Comparative virulence of Haemophilus influenzae with a type b or type d capsule. Infect Immun. 1981 May;32(2):518–524. doi: 10.1128/iai.32.2.518-524.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sansonetti P. J. Genetic and molecular basis of epithelial cell invasion by Shigella species. Rev Infect Dis. 1991 Mar-Apr;13 (Suppl 4):S285–S292. doi: 10.1093/clinids/13.supplement_4.s285. [DOI] [PubMed] [Google Scholar]
- Schiebel E., Schwarz H., Braun V. Subcellular location and unique secretion of the hemolysin of Serratia marcescens. J Biol Chem. 1989 Sep 25;264(27):16311–16320. [PubMed] [Google Scholar]
- Schulte J. M., Martich F. A., Schmid G. P. Chancroid in the United States, 1981-1990: evidence for underreporting of cases. MMWR CDC Surveill Summ. 1992 May 29;41(3):57–61. [PubMed] [Google Scholar]
- Simmons N. A. Isolation of campylobacters. Br Med J. 1977 Sep 10;2(6088):707–707. doi: 10.1136/bmj.2.6088.707-a. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sottnek F. O., Biddle J. W., Kraus S. J., Weaver R. E., Stewart J. A. Isolation and identification of Haemophilus ducreyi in a clinical study. J Clin Microbiol. 1980 Aug;12(2):170–174. doi: 10.1128/jcm.12.2.170-174.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Stevens M. K., Cope L. D., Radolf J. D., Hansen E. J. A system for generalized mutagenesis of Haemophilus ducreyi. Infect Immun. 1995 Aug;63(8):2976–2982. doi: 10.1128/iai.63.8.2976-2982.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Swihart K. G., Welch R. A. Cytotoxic activity of the Proteus hemolysin HpmA. Infect Immun. 1990 Jun;58(6):1861–1869. doi: 10.1128/iai.58.6.1861-1869.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Thompson S. A., Wang L. L., West A., Sparling P. F. Neisseria meningitidis produces iron-regulated proteins related to the RTX family of exoproteins. J Bacteriol. 1993 Feb;175(3):811–818. doi: 10.1128/jb.175.3.811-818.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Totten P. A., Lara J. C., Norn D. V., Stamm W. E. Haemophilus ducreyi attaches to and invades human epithelial cells in vitro. Infect Immun. 1994 Dec;62(12):5632–5640. doi: 10.1128/iai.62.12.5632-5640.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Totten P. A., Stamm W. E. Clear broth and plate media for culture of Haemophilus ducreyi. J Clin Microbiol. 1994 Aug;32(8):2019–2023. doi: 10.1128/jcm.32.8.2019-2023.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Uphoff T. S., Welch R. A. Nucleotide sequencing of the Proteus mirabilis calcium-independent hemolysin genes (hpmA and hpmB) reveals sequence similarity with the Serratia marcescens hemolysin genes (shlA and shlB). J Bacteriol. 1990 Mar;172(3):1206–1216. doi: 10.1128/jb.172.3.1206-1216.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wasserheit J. N. Epidemiological synergy. Interrelationships between human immunodeficiency virus infection and other sexually transmitted diseases. Sex Transm Dis. 1992 Mar-Apr;19(2):61–77. [PubMed] [Google Scholar]
- Welch R. A. Pore-forming cytolysins of gram-negative bacteria. Mol Microbiol. 1991 Mar;5(3):521–528. doi: 10.1111/j.1365-2958.1991.tb00723.x. [DOI] [PubMed] [Google Scholar]
- Willson P. J., Albritton W. L., Slaney L., Setlow J. K. Characterization of a multiple antibiotic resistance plasmid from Haemophilus ducreyi. Antimicrob Agents Chemother. 1989 Sep;33(9):1627–1630. doi: 10.1128/aac.33.9.1627. [DOI] [PMC free article] [PubMed] [Google Scholar]