1887

Journal of Medical Microbiology logo

Volume 43, Issue 3

Localisation and immunological properties of a 24-kDa surface protein of Free

Abstract

Summary

The cell wall and outer structures of bacteria were investigated. The 24-kDa outer protein from two strains was purified with an SDS–PAGE preparative continuous-elution electrophoresis cell. The protein was further characterised by SDS–PAGE and immunoblotting, and the immunological properties were investigated by ELISA. Localisation on the bacterial surface was investigated by immuno-electron-microscopy with a polyclonal antiserum raised against the purified protein. A triple-laminar cell wall typical of gram-negative bacteria, close cellular contact between bacterial cells and outer blebs were seen on thin sections. An additional high mol. wt band of . 165 kDa was seen when not treated by heating to 100°C. A high density fibrilla-like material was detected on the bacterial cell and in the environment by negative staining and immuno-electron-microscopy with antisera specific for the 24-kDa protein. The surface localisation of the 24-kDa protein was confirmed by an ELISA technique with the specific antiserum and whole bacterial cells as antigen. The presence of antibodies to the 24-kDa protein was demonstrated in antisera to 13 strains of , indicating antigenic identity or within-species cross-reactivity. Low titres of antibodies to this protein were also detected in 19 antisera raised against different strains of gram-negative bacteria, indicating cross-reactivity with other species. Antibody response to the 24-kDa protein in rabbits immunised subcutaneously with live bacteria resulted in a secondary IgG response. Of 28 sera from patients with culture-verified chancroid, 26 manifested high titres of IgG antibodies to the 24-kDa protein, thus indicating the involvement of this antigen in the disease process in man.

  • Received:
  • Accepted:
  • Published Online:
© J. MED. MICROBIOL. 1995
Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-43-3-192
1995-09-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/jmm/43/3/medmicro-43-3-192.html?itemId=/content/journal/jmm/10.1099/00222615-43-3-192&mimeType=html&fmt=ahah

References

  1. Morse S. A. Chancroid and Haemophilus ducreyi. Clin Microbiol Rev 1989; 2:137–157
     [Google Scholar]
  2. Willis S. E. Chancroid. Prim Care 1990; 17:145–152
     [Google Scholar]
  3. Cameron D. W., D’Costa L. J., Ndinya-Achola J.O., Piot P. Plummer FA. Incidence and risk factors for female to male transmission of HIV. IV International Conference on AIDS, Stockholm June, 1988. Abstract 4061
     [Google Scholar]
  4. Kreiss J. K., Coombs R., Plummer F. Isolation of human immunodeficiency virus from genital ulcers in Nairobi prostitutes. J Infect Dis 1989; 160:380–384
     [Google Scholar]
  5. Kreiss J. K., Koech D., Plummer F. A. AIDS virus infection in Nairobi prostitutes. Spread of the epidemic to East Africa. N Engl J Med 1986; 314:414–418
     [Google Scholar]
  6. Nsanze H., Fast M. V., D’Costa L. J., Tukei P., Curran J., Ronald A. R. Genital ulcers in Kenya. Clinical and laboratory study. Br J Vener Dis 1981; 57:378–381
     [Google Scholar]
  7. Lagergård T. The role of Haemophilus ducreyi bacteria, cytotoxin, endotoxin and antibodies in animal models for study of chancroid. Microb Pathog 1992; 13:203–217
     [Google Scholar]
  8. Purvén M., Lagergård T. Haemophilus ducreyi, a cytotoxin-producing bacterium. Infect Immun 1992; 60:1156–1162
     [Google Scholar]
  9. Odumeru J. A., Wiseman G. M., Ronald A. R. Relationship between lipopolysaccharide composition and virulence of Haemophilus ducreyi. J Med Microbiol 1987; 23:155–162
     [Google Scholar]
  10. 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; 50:495–499
     [Google Scholar]
  11. Jann K., Hoschützky H. Nature and organization of adhesins. Curr Top Microb Immunol 1990; 151:55–70
     [Google Scholar]
  12. Albritton W. L. Biology of Haemophilus ducreyi. Microb Rev 1989; 53:377–389
     [Google Scholar]
  13. Spinola S. M., Castellazzo A., Shero M., Apicella M. A. Characterization of pili by Haemophilus ducreyi. Microb Pathog 1990; 9:417–426
     [Google Scholar]
  14. Johnson A. P., Abeck D., Davis H. A. The structure, pathogenicity and genetics of Haemophilus ducreyi. J Infect 1988; 17:99–106
     [Google Scholar]
  15. Slootmans L., Vanden Berghe D. A., Piot P. Typing Haemophilus ducreyi by indirect immunofluorescence assay. Genitourin Med 1985; 61:123–126
     [Google Scholar]
  16. Saunders J. M., Folds J. D. Immunoblot analysis of antigens associated with Haemophilus ducreyi using serum from immunised rabbits. Genitourin Med 1986; 62:321–328
     [Google Scholar]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227:680–685
     [Google Scholar]
  18. Mattsby-Baltzer I., Lindgren K., Lindholm B., Edebo L. Endotoxin shedding by enterobacteria: free and cell-bound endotoxin differ in Limulus activity. Infect Immun 1991; 59:689–695
     [Google Scholar]
  19. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72:248–254
     [Google Scholar]
  20. Lagergård T., Purven M. Neutralizing antibodies to Haemophilus ducreyi cytotoxin. Infect Immun 1993; 61:1589–1592
     [Google Scholar]
  21. Dahlberg T., Branefors P. Enzyme-linked immunosorbent assay for titration of Haemophilus influenzae capsular and O antigen antibodies. J Clin Microbiol 1980; 12:185–192
     [Google Scholar]
  22. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76:4350–4354
     [Google Scholar]
  23. Lagergård T., Purvén M., Frisk A. Evidence of Haemophilus ducreyi adherence to and cytotoxin destruction of human epithelial cells. Microb Pathog 1993; 14:417–431
     [Google Scholar]
  24. Guerina N. G., Langermann S., Schoolnik G. K., Kessler T. W., Goldmann D. A. Purification and characterization of Haemophilus influenzae pili, and their structural and serological relatedness to Escherichia coli P and mannose-sensitive pili. J Exp Med 1985; 161:145–159
     [Google Scholar]
  25. Bakaletz L. O., Barenkamp S. J. Localization of high-molecular-weight adhesion proteins of nontypable Haemophilus influenzae by immunoelectron microscopy. Infect Immun 1994; 62:4460–4468
     [Google Scholar]
  26. Watanabe H., Kanesaki S., Shukuda Y., Mayahara H. The location of filamentous hemagglutinin and pertussis toxin antigens of Bordetella pertussis by immunoelectron microscopy. J Electron Microsc 1989; 38:172–181
     [Google Scholar]
  27. Ashworth L. A. E., Dowsett A. B., Irons L. I., Robinson A. The location of surface antigens of Bordetella pertussis by immuno-electron microscopy. Dev Biol Standard 1985; 61:143–151
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/00222615-43-3-192
Loading
Localisation and immunological properties of a 24-kDa surface protein of Haemophilus ducreyi
J. Med. Microbiol. 43, 192 (1995); https://doi.org/10.1099/00222615-43-3-192
/content/journal/jmm/10.1099/00222615-43-3-192
/content/journal/jmm/10.1099/00222615-43-3-192
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error