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Monoclonal antibodies raised against Pap fimbriae recognize minor component(s) involved in receptor binding
J M de Ree1, P Schwillens, J F van den Bosch
1Department of Medical Microbiology, University of Limburg, Maastricht, The Netherlands.
Abstract:
Pap fimbriae were purified from a recombinant strain and used for the production of monoclonal antibodies (MAbs). These MAbs were screened in a fimbriae ELISA with eight different P fimbriae as well as 1A and 1C fimbriae. Five MAbs were specific for Pap fimbriae whereas one MAb did react with Pap, F7(2) and F11 fimbriae. Previously, we described two F11 MAbs which also reacted with Pap, F7(2) and F11 fimbriae. In a whole bacteria ELISA it was shown that the MAbs, which recognized Pap, F7(2) and F11 fimbriae, reacted with recombinant strains which did not express Pap or F11 fimbriae, but still expressed the globoside binding properties. Not one of the five MAbs which are specific for Pap fimbriae reacted with these globoside binding recombinant strains. In a haemagglutination and adherence assay it was shown that only the MAbs which recognized the Pap, F7(2) and F11 fimbriae inhibited the adhesive properties of the globoside binding recombinant strain. Therefore it is concluded that in the present study MAbs are presented which recognize the minor components responsible for adhesion.
Insights
Monoclonal antibodies (MAbs) targeting Pap fimbriae were developed. These MAbs identified minor components responsible for bacterial adhesion, aiding in understanding fimbrial function.
Area of Science:
- Microbiology
- Immunology
- Bacterial Pathogenesis
Background:
- Pap fimbriae are crucial virulence factors in uropathogenic Escherichia coli.
- Understanding the specific components of fimbriae responsible for adhesion is vital for developing targeted therapies.
Purpose of the Study:
- To produce and characterize monoclonal antibodies (MAbs) against Pap fimbriae.
- To identify MAbs that recognize specific Pap fimbrial components involved in bacterial adhesion.
Main Methods:
- Purification of Pap fimbriae from a recombinant strain.
- Production and screening of monoclonal antibodies using enzyme-linked immunosorbent assays (ELISA).
- Functional assays including haemagglutination and adherence assays.
Main Results:
- Five MAbs were specific for Pap fimbriae.
- One MAb cross-reacted with Pap, F7(2), and F11 fimbriae.
- MAbs recognizing Pap, F7(2), and F11 fimbriae inhibited globoside binding and adhesion, indicating they target minor adhesive components.
Conclusions:
- The study presents novel MAbs that recognize minor components responsible for bacterial adhesion.
- These findings contribute to a deeper understanding of Pap fimbriae structure-function relationships.
- The identified MAbs could be valuable tools for diagnostics and therapeutic interventions targeting bacterial infections.