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United States Department of Agriculture

Agricultural Research Service

Research Project: GENOMIC AND IMMUNOLOGICAL CHARACTERISTICS OF JOHNE'S DISEASE Title: Immune Responses in Mice to Mycobacterium avium subsp. paratuberculosis Following Vaccination with a Novel 74F Recombinant Polyprotein

item Chen, Li-Hsuen - CORNELL UNIVERSITY
item Kathaperumal, Kumanan - CORNELL UNIVERSITY
item Huang, Ching-Juo - CORNELL UNIVERSITY
item Mcdonough, Sean - CORNELL UNIVERSITY
item Stehman, Susan - CORNELL UNIVERSITY
item Huntley, John - NY STATE DEPT OF AG.
item Bannantine, John
item Chang, Chao-Fu - CORNELL UNIVERSITY
item Chang, Yung-Fu - CORNELL UNIVERSITY

Submitted to: Vaccine
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: December 13, 2007
Publication Date: January 13, 2008
Repository URL:
Citation: Chen, L., Kathaperumal, K., Huang, C., Mcdonough, S.P., Stehman, S., Akey, B., Huntley, J., Bannantine, J.P., Chang, C., Chang, Y. 2008. Immune Responses in Mice to Mycobacterium avium subsp. paratuberculosis Following Vaccination with a Novel 74F Recombinant Polyprotein. Vaccine. 26(9):1253-1262.

Interpretive Summary: In this communication, we merged three separate proteins produced by Mycobacterium avium subsp paratuberculosis into a single, long protein called a polyprotein (termed 74F in this study). This was done using recombinant DNA techniques. The three proteins have been shown to elicit protective responses in animals against TB infection, and therefore, the hypothesis was that these proteins might also be protective against infection with Mycobacterium avium subsp paratuberculosis, the bacterium that causes Johne’s disease. We evaluated protection using mice and showed that mouse tissues did have fewer lesions and bacteria load when compared to control mice.

Technical Abstract: Johne’s Disease (JD) is a chronic infectious disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Here, we report the cloning and expression of a 74kDa recombinant polyprotein (Map74F) and its protective efficacy against MAP infection in mice. Map74F was generated by the sequential linkage of the ORFs of the~17.6-kDa C-terminal fragment of Map3527 to the full-length ORF of Map1519, followed at the C terminus with ~14.6-kDa N-terminal portion of Map3527. Mice immunized with Map74F had a significant IgG1 response but not IgG2a. In immunized animals, the IgG1/IgG2a ratio increased until 4 wk after MAP challenge. The ratio decreased from 8 wk indicating a shift to a Th1 response. Antigen specific IFN-gamma response, CD3+ and CD4+ T cells increased significantly in immunized mice. Following challenge, MAP burden was significantly lower in liver, spleen and mesenteric lymph nodes of immunized animals compared to control animals indicating protection against MAP infection. This was further evident by the improved liver and spleen pathology of the immunized animals, which had fewer granulomas and lower numbers of acid-fast bacilli. Results of this study indicated that immunization of mice with Map74F protected mice against MAP infection.

Last Modified: 11/28/2015
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