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Clinical and Vaccine Immunology, June 2006, p. 611-619, Vol. 13, No. 6
1071-412X/06/$08.00+0     doi:10.1128/CVI.00054-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Immune Responses to Defined Antigens of Mycobacterium bovis in Cattle Experimentally Infected with Mycobacterium kansasii

United States Department of Agriculture, Agricultural Research Service, National Animal Disease Center, Bacterial Diseases of Livestock Research Unit, Ames, Iowa,¹ USDA, Animal and Plant Health Inspection Service (APHIS), Veterinary Services, National Veterinary Services Laboratory, Mycobacteria and Brucella Section, Ames, Iowa 50010,² Iowa State University, College of Veterinary Medicine, Department of Veterinary Microbiology and Preventive Medicine, Ames, Iowa 50011,³ Chembio Diagnostic Systems, Inc., Medford, New York,⁴ Statens Serum Institut, Copenhagen, Denmark,⁵ Bacteriology Department, Veterinary Sciences Division, Stormont, Belfast, United Kingdom⁶

Received 10 February 2006/ Returned for modification 21 March 2006/ Accepted 24 March 2006

Cross-reactive responses elicited by exposure to nontuberculous mycobacteria often confound the interpretation of antemortem tests for Mycobacterium bovis infection of cattle. The use of specific proteins (e.g., ESAT-6, CFP-10, and MPB83), however, generally enhances the specificity of bovine tuberculosis tests. While genes for these proteins are absent from many nontuberculous mycobacteria, they are present in M. kansasii. Instillation of M. kansasii into the tonsillar crypts of calves elicited delayed-type hypersensitivity and in vitro gamma interferon and nitrite concentration responses of leukocytes to M. avium and M. bovis purified protein derivatives (PPDs). While the responses of M. kansasii-inoculated calves to M. avium and M. bovis PPDs were approximately equivalent, the responses of M. bovis-inoculated calves to M. bovis PPD exceeded their respective responses to M. avium PPD. The gamma interferon and nitrite responses of M. kansasii-inoculated calves to recombinant ESAT-6-CFP-10 (rESAT-6-CFP-10) exceeded corresponding responses of noninoculated calves as early as 15 and 30 days after inoculation, respectively, and persisted throughout the study. The gamma interferon and nitrite responses of M. bovis-inoculated calves to rESAT-6-CFP-10 exceeded the corresponding responses of M. kansasii-inoculated calves beginning 30 days after inoculation. By using a lipoarabinomannan-based enzyme-linked immunosorbent assay, specific serum antibodies were detected as early as 50 days after challenge with M. kansasii. By a multiantigen print immunoassay and immunoblotting, serum antibodies to MPB83, but not ESAT-6 or CFP-10, were detected in M. kansasii-inoculated calves; however, responses to MPB83 were notably weaker than those elicited by M. bovis infection. These findings indicate that M. kansasii infection of calves elicits specific responses that may confound the interpretation of bovine tuberculosis tests.

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