Assessment by electron-microscopy of recombinant Vibrio cholerae and Salmonella vaccine strains expressing enterotoxigenic Escherichia coli-specific surface.

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Assessment by electron-microscopy of recombinant Vibrio cholerae and Salmonella vaccine strains expressing enterotoxigenic Escherichia coli-specific surface antigens  V. Ziethlow, D. Favre, J.-F. Viret, J. Frey, M.H. Stoffel  Clinical Microbiology and Infection  Volume 14, Issue 3, Pages 282-286 (March 2008) DOI: 10.1111/j.1469-0691.2007.01908.x Copyright © 2008 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 1 Western blot assays showing reactivity of mouse monoclonal anti-CFA/I, anti-CS3 and anti-CS6 antibodies with laboratory and wild-type strains, with native and recombinant host strains, and with purified antigen. Clinical Microbiology and Infection 2008 14, 282-286DOI: (10.1111/j.1469-0691.2007.01908.x) Copyright © 2008 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 2 Analysis by electron-microscopy using negative staining and immunogold labelling following addition of monoclonal anti-CFA/I antibodies. (a) Labelling of the wild-type strain H10407. Absence of labelling of the native host strains (b) CVD103 and (c) Salmonella Ty21a substantiates the suitability of the detection system. (d) Recombinant monovalent strain CVD103-CFA/I did not exhibit any immunogold labelling, but (e) CFA/I pili were visualised at the surface of Salmonella-CFA/I by negative staining and immunogold labelling. (f–j) Corresponding strains after labelling with anti-CS3 antibodies: (f) labelling of wild-type strain DS198–1 and the absence of signal on the native host strains (g) CVD103-HgR and (h) Salmonella Ty21a corroborates the reliability of the detection system. CS3 pili were visualised at the surface of the monovalent recombinant strains (i) CVD-HgR-CS3 and (j) Salmonella-CS3. (k–m) Control experiments: apart from a few occasional gold particles, no signal was observed when wild-type strains were labelled with (k) mouse monoclonal anti-osteoprotegerin or (l, m) mouse monoclonal anti-collagen IV as inappropriate primary antibodies. Scale bar = 0.5 μm. Clinical Microbiology and Infection 2008 14, 282-286DOI: (10.1111/j.1469-0691.2007.01908.x) Copyright © 2008 European Society of Clinical Infectious Diseases Terms and Conditions