This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Dertzbaugh, M T Articles by Elson, C O Search for Related Content PubMed PubMed Citation Articles by Dertzbaugh, M T Articles by Elson, C O

 Previous Article  |  Next Article 

Infect Immun. 1993 February; 61(2): 384-390

Reduction in oral immunogenicity of cholera toxin B subunit by N-terminal peptide addition.

Division of Gastroenterology, School of Medicine, University of Alabama, Birmingham 35294.

ABSTRACT

The mucosal adjuvanticity of cholera toxin and the potential of the B subunit of cholera toxin (CtxB) to serve as an oral vaccine carrier have prompted interest in the coupling of immunogenic peptides to this protein. The purpose of this study was to determine how such fusions affect the function of CtxB. Oligonucleotides were genetically fused to the 5' terminus of the ctxB gene to encode additional amino acids of 8, 12, and 24 residues in length. None of these additions affected the ability of CtxB to oligomerize, as determined by nondenaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Circular dichroism revealed no difference in conformation between the modified B subunits, regardless of the length of the addition. However, when compared with native CtxB, additions to the N terminus induced a consistent change in the net conformation of the protein. By using a competitive enzyme immunoassay, the affinity of the modified B subunits for GM1 ganglioside was shown to gradually decrease with increasing length of the N-terminal addition. A similar pattern was observed for the ability of the chimeras to inhibit proliferation of concanavalin A-stimulated spleen cells in vitro, which is a previously described functional property of CtxB that is dependent on its binding to cells. Lastly, the oral immunogenicity of these chimeras was found to be less than that of native CtxB. These results indicate that large fusions to the N terminus of CtxB can significantly affect its biological properties and could reduce its value as a mechanism for effective mucosal immunization.

This article has been cited by other articles:

• Hajishengallis, G., Arce, S., Gockel, C.M., Connell, T.D., Russell, M.W. (2005). Immunomodulation with Enterotoxins for the Generation of Secretory Immunity or Tolerance: Applications for Oral Infections. J. Dent. Res. 84: 1104-1116 [Abstract] [Full Text]  
• Sanchez, A. E., Aquino, G., Ostoa-Saloma, P., Laclette, J. P., Rocha-Zavaleta, L. (2004). Cholera Toxin B-Subunit Gene Enhances Mucosal Immunoglobulin A, Th1-Type, and CD8+ Cytotoxic Responses When Coadministered Intradermally with a DNA Vaccine. CVI 11: 711-719 [Abstract] [Full Text]  
• Harokopakis, E., Hajishengallis, G., Michalek, S. M. (1998). Effectiveness of Liposomes Possessing Surface-Linked Recombinant B Subunit of Cholera Toxin as an Oral Antigen Delivery System. Infect. Immun. 66: 4299-4304 [Abstract] [Full Text]  
• Hajishengallis, G., Russell, M. W., Michalek, S. M. (1998). Comparison of an Adherence Domain and a Structural Region of Streptococcus mutans Antigen I/II in Protective Immunity against Dental Caries in Rats after Intranasal Immunization. Infect. Immun. 66: 1740-1743 [Abstract] [Full Text]  
• Sultan, F., Jin, L.-l., Jobling, M. G., Holmes, R. K., Stanley, S. L. Jr. (1998). Mucosal Immunogenicity of a Holotoxin-Like Molecule Containing the Serine-Rich Entamoeba histolytica Protein (SREHP) Fused to the A2 Domain of Cholera Toxin. Infect. Immun. 66: 462-468 [Abstract] [Full Text]