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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Infectious Bacterial Diseases Research » Research » Publications at this Location » Publication #279956

Title: MAP1272c encodes a NlpC/P60 protein, an antigen detected in cattle with Johne's Disease

item Bannantine, John
item LINGLE, CARI - University Of Missouri
item Stabel, Judith
item RAMYAR, KASRA - University Of Missouri
item GARCIA, BRANDON - University Of Missouri
item RAEBER, ALEX - Prionics Ag
item SCHACHER, PASCAL - Prionics Ag
item KAPUR, VIVEK - Pennsylvania State University
item GEISBRECHT, BRIAN - University Of Missouri

Submitted to: Clinical and Vaccine Immunology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/11/2012
Publication Date: 5/16/2012
Citation: Bannantine, J.P., Lingle, C.K., Stabel, J.R., Ramyar, K.X., Garcia, B.L., Raeber, A., Schacher, P., Kapur, V., Geisbrecht, B.V. 2012. MAP1272c encodes a NlpC/P60 protein, an antigen detected in cattle with Johne's Disease. Clinical and Vaccine Immunology. 19(7):1083-1092.

Interpretive Summary: Johne’s disease in dairy cattle costs millions of dollars in lost production for the US dairy industry alone. The disease could be better controlled through the use of improved diagnostic tests to detect Mycobacterium avium subspecies paratuberculosis (MAP), the bacterium that causes Johne’s disease. In an effort to develop an improved diagnostic test, the USDA partnered with Prionics AG to analyze a set of MAP proteins to find which ones showed the best reaction with sera from cattle with Johne’s disease. One of the proteins, encoded by MAP1272c, clearly stood out as being the best protein to use in an antigen-based test such as an ELISA. Monoclonal antibodies were made against this protein and the protein was then compared with a commercially available ELISA test. It was discovered that the commercially available test still worked better at detecting Johne’s disease. This information will enable veterinarians to make informed choices about diagnostic tests to use for herds they manage.

Technical Abstract: The protein encoded by MAP1272c has been shown to be an antigen of Mycobacterium avium subspecies paratuberculosis that contains an NlpC/P60 superfamily domain found in lipoproteins or integral membrane proteins. Proteins containing this domain have diverse enzymatic functions that include peptidases, amidases and acetyltransferases. The NlpC protein was further examined in comparison to 95 other recombinant proteins and showed the strongest reactivity when analyzed with sera from Johne’s disease cattle. To further localize the immunogenicity of NlpC, recombinant proteins representing defined regions were expressed and evaluated with sera from cattle with Johne’s disease. The region from amino acids 74-279 was shown to be the most immunogenic. This fragment was also evaluated against a commercially available ELISA test. Two monoclonal antibodies were produced in mice immunized with the full-length protein and each recognized a distinct epitope. These antibodies cross-reacted with proteins from other mycobacterial species and demonstrated variable sizes of the proteins expressed from these subspecies. The hypervariable loops of both antibodies were sequenced and their interaction with MAP1272c was characterized by solution-based, luminescent binding assay. These tools provide additional means to study a strong antigen of M. avium subsp paratuberculosis.