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E. coli 0157:147 Intimin Expressed by Transgenic Plant Cells as a Candidate Oral Vaccine for Cattle


The ultimate goal of this investigation is to develop an inexpensive, effective, easily administered vaccine to prevent cattle from becoming infected with E. coli 0157:147 so as to block transmission of these organisms to humans. The carboxy-terminal portion of intimino157 of E. coli 0157:147 is being used as the immunogen for such a vaccine because: i.) antibodies against this portion of the outer membrane protein adhesin block adherence of the microbe to tissue culture cells and in a mouse model of colonization, ii.) this portion of intimino157 has been plant optimized to increase expression of the protein in plant expression systems.

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The project was briefly delayed when it was discovered that the carboxy-terminal portion of intimino157 was being glycosylated in the plant cell system. In E. coli 0157:147, intimino157 is not glycosylated. The gylcosylation of the protein was the result of an aberrant immune response that was unable to block colonization of E. coli 0157:147. This problem has been remedied and a new plant cell clone expressing an unglycosylated carboxy-terminal portion of intimino157 is currently being characterized. In collaboration with Dr. Wayne Curtis of Penn State University, production of the plant cell clone expressing the plant-optimized carboxy-terminal portion of intimino 157 is being scaled-up. Previous studies have shown that mice fed plant cells expressing intimino157 make an antibody response that blocks adherence of E. coli 0157:147 to tissue culture cells. In addition, lab results have shown that mice immunized intraperitoneally with purified carboxy-terminal portion of intimino 157 are not colonized to the same extent as non-immunized mice. These results suggest that an orally-delivered vaccine comprised of the carboxy-terminal portion of intimino 157 can function in blocking adherence of E. coli 0157:147 in the mouse model of E. coli 0157:147 colonization.

O'Brien, Alison
Uniformed Services University of the Health Sciences
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