Cross-Reactive Monoclonal Antibodies Induced by Lps from Periodontopathic Bacteria

Abstract

Monoclonal antibodies against lipopolysaccharides (LPS) of Fusobacterium nucleatum ATCC 25586 and Eikenella corrodens FDC 1073 were used for the study of antigenic characteristics of periodontopathic bacteria.

LPS was extracted by the hot-phenol/water method. Three monoclonal antibodies (isotype IgM) against F. nucleatum ATCC 25586 were cross-reactive with LPS from E. corrodens FDC 1073. Four hybridomas producing antibody to E. corrodens FDC 1073 LPS were also obtained. Three out of four monoclonal antibodies (isotype IgM) were found to exhibit cross-reactivity with LPS from F. nucleatum ATCC 25586. These six monoclonal antibodies reacted with four out of seven E. corrodes and nine out of 27 F. nucleatum clinical isolates. Western immunoblotting analysis demonstrated that the six antibodies reacted with a similar fast-migrating band of LPS from F. nucleatum ATCC 25586 and E. corrodens FDC 1073. Another monoclonal antibody (isotype IgG3) against E. corrodens FDC 1073 LPS exhibited cross-reactivity with LPS from Fusobacterium necrophorum ATCC 25286 and Capnocytophaga ochracea M-12. This antibody was found to react with slow-migrating bands of E. corrodens FDC 1073 LPS and a fast-migrating band of LPS from F. necrophorum ATCC 25286 and C. ochracea M-12 by Western immunoblotting analysis. Four out of seven clinical isolates of E. corrodens reacted strongly with this antibody.

The present findings indicate that there is a common LPS core epitope between F. nucleatum ATCC 25586 and E. corrodens FDC 1073. They also indicated that the O-side-chain epitope of E. corrodens FDC 1073 might be common to the LPS core epitopes of F. necrophorum ATCC 25286 and C. ochracea M-12.

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References

Aadnegard, B. and Hofstad, T. (1983): O-Antigenic Cross-reactivity in Fusobacterium nucleatum: Chemotype V Lipopolysaccharides, Acta Pathol Microbiol Scand (Sect B) 91: 93-95.

Darwish, S. ; Hyppa, T. ; and Socransky, S.S. (1978): Studies of the Predominant Cultivable Microbiota of Early Periodontitis, J Periodont Res 13: 1-16.

Galanos, C. ; Rietschel, E.T. ; Luderitz, O. ; and Westphal, O. (1971): Interaction of Lipopolysaccharides and Lipid A with Complement, Eur J Biochem 19: 143-152.

Hase, S. ; Hofstad, T. ; and Rietschel, T. (1977): Chemical Structure of the Lipid A Component of Lipopolysaccharides from Fusobacterium nucleatum, J Bacteriol 129: 9-14.

Hofstad, T. ; Skaug, N. ; and Bjornland, T. (1979): O-Antigenic Cross-reactivity in Fusobacterium nucleatum, Acta Pathol Microbiol Scand (Sect B) 87:371-374.

Laemmli, U.K. (1970): Cleavage of Structural Proteins During the Assembly of the Head of Bacteriophage T4, Nature (London) 227: 680-685.

Mashimo, J. ; Yoshida, M. ; Ikeuchi, K. ; Hata, S. ; Arata, S. ; Kasai, N. ; Okuda, K. ; and Takazoe, I. (1985): Fatty Acid Composition and Shwartzman Activity of Lipopolysaccharides from Oral Bacteria, Microbiol Immunol 29: 395-403.

Mutharia, L.M. ; Crockford, G. ; Bogard, W.C., Jr. ; and Hancock, R.E.W. (1984): Monoclonal Antibodies Specific for Escherichia coli J5 Lipopolysaccharide: Cross-reaction with Other Gram-negative Bacterial Species, Infect Immun 45: 631-636.

Nelles, M.J. and Niswander, C.A. (1984): Mouse Monoclonal Antibodies Reactive with J5 Lipopolysaccharide Exhibit Extensive Serological Cross-reactivity with a Variety of Gram-negative Bacteria, Infect Immun 46: 677-681.

10.

Okuda, K. ; Naito, Y. ; Fukumoto, Y. ; Kimura, Y. ; Ishi-Kawa, I. ; Kinoshita, S. ; and Takazoe, I. (1984): Bacteriological Study of Periodontal Lesions in Two Sisters with Juvenile Periodontitis and Their Mother, Infect Immun 45: 118-121.

11.

Progulske, A. and Holt, S.C. (1984): Isolation and Characterization of the Outer Membrane and Lipopolysaccharide from Eikenella corrodens, Infect Immun 43: 166-177.

12.

Progulske, A. ; Mishell, R. ; Trummel, C. ; and Holt, S.C. (1984): Biological Activities of Eikenella corrodens Outer Membrane and Lipopolysaccharide, Infect Immun 43: 178-182.

13.

Slots, J. (1977): The Predominant Cultivable Microflora of Advanced Periodontitis, Scand J Dent Res 85: 114-121.

14.

Socransky, S.S. (1977): Microbiology of Periodontal Disease-Present Status and Future Considerations, J Periodontol 48: 497-504.

15.

Tanner, A.C.R. ; Haffer, C. ; Bratthall, R.A. ; Visconti, R.A. ; and Socransky, S.S. (1979): A Study of the Bacteria Associated with Advancing Periodontitis in Man, J Clin Periodontol 6: 278-307.

16.

Towbin, H. ; Staehelin, T. ; and Gordon, J. (1979): Electrophoretic Transfer of Proteins from Polyacrylamide Gels to Nitrocellulose Sheets: Procedure and Some Applications, Proc Natl Acad Sci USA 76: 4350-4354.

17.

Westphal, O. and Jann, K. (1965): Bacterial Lipopolysaccharide. Extraction with Phenol-Water and Further Applications of the Procedures, Meth Carbohydr Chem 5: 83-91.