Diagnostic Implications of Antigen-Induced Gamma Interferon, Nitric Oxide, and Tumor Necrosis Factor Alpha Production by Peripheral Blood Mononuclear Cells from Mycobacterium bovis-Infected Cattle

Authors

W. R. Waters, United States Department of Agriculture, Agricultural Research Service, National Animal Disease Center, Ames, Iowa
M. V. Palmer, United States Department of Agriculture, Agricultural Research Service, National Animal Disease Center, Ames, IowaFollow
D. L. Whipple, United States Department of Agriculture, Agricultural Research Service, National Animal Disease Center, Ames, Iowa
M. P. Carlson, University of Nebraska Veterinary Diagnostic Center, Lincoln, Nebraska
B. J. Nonnecke, University of Nebraska Veterinary Diagnostic Center, Lincoln, Nebraska

Date of this Version

9-1-2003

Comments

Published in CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY, Sept. 2003, p. 960–966.

Abstract

Bovine tuberculosis in the United States has proven costly to cattle producers as well as to government regulatory agencies. While in vivo responsiveness to mycobacterial antigens is the current standard for the diagnosis of tuberculosis, in vitro assays are gaining acceptance, especially as ancillary or complementary tests. To evaluate in vitro indices of cellular sensitization, antigen-induced gamma interferon (IFN-γ), nitric oxide (NO), and tumor necrosis factor alpha (TNF-α) responses by blood mononuclear cells from Mycobacterium bovis-infected cattle were quantified and compared. Using an aerosol model of infection, two doses of each of two strains of M. bovis (95-1315 and HC-2045T) were used to induce a range of IFN-γ, NO, and TNF-α responses. Infection-specific increases in NO, but not in IFN-γ or TNF-α, were detected in nonstimulated cultures at 48 h, a finding that is indicative of nonspecific activation and spontaneous release of NO. The infective dose of M. bovis organisms also influenced responses. At 34 days postinfection, IFN-γ, NO, and TNF-α responses in antigen-stimulated cells from cattle receiving 105 CFU of M. bovis organisms were greater than responses of cells from cattle infected with 103 CFU of M. bovis organisms. The NO response, but not the IFN-γ and TNF-α responses, was influenced by infective strains of M. bovis. The TNF-α, NO, and IFN-γ responses followed similar kinetics, with strong positive associations among the three readouts. Overall, these findings indicate that NO and TNF-α, like IFN-γ, may prove useful as indices for the diagnosis of bovine tuberculosis.