This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow E-mail this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ASM journals
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Gordon, V M
Right arrow Articles by Samuel, J E
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Gordon, V M
Right arrow Articles by Samuel, J E

 Previous Article  |  Next Article 

Infect Immun. 1992 February; 60(2): 485-490

An enzymatic mutant of Shiga-like toxin II variant is a vaccine candidate for edema disease of swine.

V M Gordon, S C Whipp, H W Moon, A D O'Brien and J E Samuel

Department of Microbiology, Uniformed Services University of the Health Sciences, Bethesda, Maryland 22908.

ABSTRACT

Edema disease (ED) of weanling pigs is caused by an infection with Escherichia coli that produces Shiga-like toxin II variant (SLT-IIv). Pathology identical to that caused by ED can be duplicated in pigs that are injected with less than 10 ng of purified SLT-IIv per kg of body weight. Therefore, SLT-IIv was mutated to create an immunoreactive form of the toxin that was significantly reduced in enzymatic activity. Initially, purified SLT-IIv was treated with formaldehyde which abrogated cytotoxic activity. Pigs were vaccinated with the toxoid (100 micrograms) to determine whether a toxoid was a viable vaccine candidate and whether young pigs were capable of mounting an immune response. Although the pigs developed a neutralizing antibody titer (1:128 to 1:512) 28 days postinjection, they also lost weight and developed ED lesions. The deleterious effect of the toxoid appeared to result from residual enzymatic activity or a reversion to a toxic form. An alternative method, site-directed mutagenesis, was employed to consistently reduce the enzymatic activity of SLT-IIv. Glutamate at position 167 of the mature A subunit was replaced by aspartate (E167D), and arginine at position 170 was replaced by lysine (R170K). These mutations reduced cytotoxic activity 10(4)-fold and 10-fold, respectively, while the enzymatic activities were decreased 400-fold and 5-fold, respectively. The activity of a toxin that contained both mutations (SLT-IIvE167D/R170K) closely resembled that of SLT-IIvE167D. When position 167 was replaced by glutamine (E167Q), the cytotoxic activity decreased 10(6)-fold and the enzymatic activity decreased approximately 1,500-fold. Pigs that were vaccinated with purified, mutant toxin designated SLT-IIvE167Q developed a neutralizing antibody titer of 1:512 21 days postinjection, and their tissues were free of ED lesions. These data suggest that SLT-IIvE167Q may represent an effective vaccine against ED.

Infect Immun. 1992 February; 60(2): 485-490




This article has been cited by other articles:

  • Bentancor, L. V., Bilen, M., Brando, R. J. F., Ramos, M. V., Ferreira, L. C. S., Ghiringhelli, P. D., Palermo, M. S. (2009). A DNA Vaccine Encoding the Enterohemorragic Escherichia coli Shiga-Like Toxin 2 A2 and B Subunits Confers Protective Immunity to Shiga Toxin Challenge in the Murine Model. CVI 16: 712-718 [Abstract] [Full Text]  
  • Hoffman, M. A., Menge, C., Casey, T. A., Laegreid, W., Bosworth, B. T., Dean-Nystrom, E. A. (2006). Bovine Immune Response to Shiga-Toxigenic Escherichia coli O157:H7. CVI 13: 1322-1327 [Abstract] [Full Text]  
  • Marcato, P., Griener, T. P., Mulvey, G. L., Armstrong, G. D. (2005). Recombinant Shiga Toxin B-Subunit-Keyhole Limpet Hemocyanin Conjugate Vaccine Protects Mice from Shigatoxemia. Infect. Immun. 73: 6523-6529 [Abstract] [Full Text]  
  • Talbot, U. M., Paton, J. C., Paton, A. W. (2005). Protective Immunization of Mice with an Active-Site Mutant of Subtilase Cytotoxin of Shiga Toxin-Producing Escherichia coli. Infect. Immun. 73: 4432-4436 [Abstract] [Full Text]  
  • Matise, I., Cornick, N. A., Samuel, J. E., Moon, H. W. (2003). Binding of Shiga Toxin 2e to Porcine Erythrocytes In Vivo and In Vitro. Infect. Immun. 71: 5194-5201 [Abstract] [Full Text]  
  • Capozzo, A. V. E., Creydt, V. P., Dran, G., Fernandez, G., Gomez, S., Bentancor, L. V., Rubel, C., Ibarra, C., Isturiz, M., Palermo, M. S. (2003). Development of DNA Vaccines against Hemolytic-Uremic Syndrome in a Murine Model. Infect. Immun. 71: 3971-3978 [Abstract] [Full Text]  
  • Ishikawa, S., Kawahara, K., Kagami, Y., Isshiki, Y., Kaneko, A., Matsui, H., Okada, N., Danbara, H. (2003). Protection against Shiga Toxin 1 Challenge by Immunization of Mice with Purified Mutant Shiga Toxin 1. Infect. Immun. 71: 3235-3239 [Abstract] [Full Text]  
  • Cornick, N. A., Booher, S. L., Casey, T. A., Moon, H. W. (2000). Persistent Colonization of Sheep by Escherichia coli O157:H7 and Other E. coli Pathotypes. Appl. Environ. Microbiol. 66: 4926-4934 [Abstract] [Full Text]  
  • Foster, G. H., Armstrong, C. S., Sakiri, R., Tesh, V. L. (2000). Shiga Toxin-Induced Tumor Necrosis Factor Alpha Expression: Requirement for Toxin Enzymatic Activity and Monocyte Protein Kinase C and Protein Tyrosine Kinases. Infect. Immun. 68: 5183-5189 [Abstract] [Full Text]  
  • Paton, J. C., Paton, A. W. (1998). Pathogenesis and Diagnosis of Shiga Toxin-Producing Escherichia coli Infections. Clin. Microbiol. Rev. 11: 450-479 [Abstract] [Full Text]  


Copyright © 2010 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»