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Research Article

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

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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, J E Samuel
Infection and Immunity Feb 1992, 60 (2) 485-490; DOI:

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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, J E Samuel
Infection and Immunity Feb 1992, 60 (2) 485-490; DOI:
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