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

Waters, W. Ray; Palmer, M. V.; Whipple, Diana L.; Carlson, Mats; Nonnecke, Brian J.

doi:10.1128/cdli.10.5.960-966.2003

Cited by 32 publications

(31 citation statements)

References 38 publications

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“…Irrelevant control peptides were synthesized by New England Peptide and used at a concentration of 1 mg/ml. Whole-cell sonicates (WCSs) and proteinase K-digested WCSs (PKWCSs) from M. bovis 95-1315 were used at 10 mg/ml (44). WCSs from L. borgpetersenii serovar Hardjo, Brucella ovis, and Treponema phagedenis were used at 10 mg/ml as negative-control Ags (45,46).…”

Section: Abs and Reagentsmentioning

confidence: 99%

Specific Recognition of Mycobacterial Protein and Peptide Antigens by γδ T Cell Subsets following Infection with VirulentMycobacterium bovis

McGill

Sacco

Baldwin

et al. 2014

The Journal of Immunology

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Promoting effective immunity to Mycobacterium bovis infection is a challenge that is of interest to the fields of human and animal medicine alike. We report that γδ T cells from virulent M. bovis–infected cattle respond specifically and directly to complex, protein, and nonprotein mycobacterial Ags. Importantly, to our knowledge, we demonstrate for the first time that bovine γδ T cells specifically recognize peptide Ags derived from the mycobacterial protein complex ESAT6:CFP10 and that this recognition requires direct contact with APCs and signaling through the T cell Ag receptor but is independent of MHC class I or II. Furthermore, we show that M. bovis infection in cattle induces robust IL-17A protein responses. Interestingly, in contrast to results from mice, bovine CD4 T cells, and not γδ T cells, are the predominant source of this critical proinflammatory mediator. Bovine γδ T cells are divided into subsets based upon their expression of Workshop Cluster 1 (WC1), and we demonstrate that the M. bovis–specific γδ T cell response is composed of a heterogeneous mix of WC1-expressing populations, with the serologically defined WC1.1+ and WC1.2+ subsets responding in vitro to mycobacterial Ags and accumulating in the lesions of M. bovis–infected animals. The results described in this article enhance our understanding of γδ T cell biology and, because virulent M. bovis infection of cattle represents an excellent model of tuberculosis in humans, contribute to our overall understanding of the role of γδ T cells in the mycobacterial-specific immune response.

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Section: Abs and Reagentsmentioning

confidence: 99%

Specific Recognition of Mycobacterial Protein and Peptide Antigens by γδ T Cell Subsets following Infection with VirulentMycobacterium bovis

McGill

Sacco

Baldwin

et al. 2014

The Journal of Immunology

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“…All animal care and use procedures were reviewed and approved by the NADC Animal Care and Use Committee. Calves received M. bovis strain 95-1315 by aerosol at 6 months of age as described previously (12). Blood samples for this study were collected 4 months after infection.…”

Section: Animalsmentioning

confidence: 99%

Optimization of a Whole-Blood Gamma Interferon Assay for Detection of Mycobacterium bovis -Infected Cattle

Schiller¹,

Waters

Vordermeier

et al. 2009

Clin Vaccine Immunol

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Antigens of Mycobacterium bovis elicit a cell-mediated immune response upon intradermal injection in cattle.In vitro, such antigens stimulate the production of gamma interferon (IFN-␥) by bovine T cells in whole-blood culture (IFN-␥ assay). We have analyzed various parameters of the in vitro IFN-␥ assay, ranging from blood sampling to execution of the IFN-␥ test, in view of potential simplifications of the assay. Here, we show that IFN-␥ responses may be reduced under certain animal handling/holding conditions and that a delayed time from blood collection to culture may lead to a reduced in vitro IFN-␥ response. Delayed initiation of culture in a purified-protein-derivative-based assay (24 h compared to 8 h after blood collection), however, resulted in a significant improvement of specificity (97% compared to 85%), whereas there was only a modest reduction of sensitivity (from 96% to 90%), which was statistically not significant. Furthermore, we show that the stimulation temperature needs to be 33°C or higher; that carbon dioxide is not required for stimulation; and that various plate formats, ranging from 24 to 96 wells per plate, can be utilized. The produced IFN-␥ is stable at 4°C for 28 days as well as after repeated freeze-thaw cycles. Thus, stimulation of samples may be initiated in the field without the need for a carbon dioxide source, and bovine IFN-␥ is stable under various routine laboratory temperature scenarios. These findings demonstrate opportunities for improvements in the bovine IFN-␥ test platform and flexibilities in test application.Bovine tuberculosis (TB), caused by Mycobacterium bovis, has an important and adverse effect on socioeconomic conditions, public health, and trade of animals and animal products (2). Eradication of bovine TB in cattle is based on detection and slaughter of infected animals or whole herds. The standard antemortem screening test for detection of TB is the intradermal tuberculin skin test (i.e., intradermal injection of tuberculin eliciting a cell-mediated immune response [CMI] at the site, which in turn leads to skin thickening). As an alternative, the CMI can be measured in vitro by stimulating blood cells with tuberculin, which in turn leads to production of gamma interferon (IFN-␥), which can then be quantified by an enzyme-linked immunosorbent assay (ELISA; Bovigam IFN-␥ assay) (15).The Bovigam assay constitutes a laboratory-based TB test and is widely used complementarily to the tuberculin skin test (4, 11), as it offers national TB control programs and industry an additional tool for curtailing the spread of TB in cattle and other Bovidae. The assay critically depends on the sample quality, culture conditions, and quality control of stimulation reagents. The CMI, both in vivo and in vitro, may be negatively affected by stress or corticosteroid application (5). Thus, parallel stimulation of blood leukocytes with mitogen or superantigen in the IFN-␥ assay is commonly used as an indicator of sample quality and potential for underlying CMI suppression, thereby re...

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“…Diferentes autores han demostrado que esta prueba es más sensible y especifica que la prueba intradérmica (17,21,22) . Sin embargo, la exposición a otras micobacterias o la co-infección con M. avium subsp paratuberculosis, afecta la especificidad de las pruebas basadas en el uso del PPD, incrementando el número de reacciones falsas positivas, fenómeno que puede ser evitado usando antígenos altamente inmunogénicos como ESAT-6 y CFP-10 (23) .…”

Section: Cuadro 1 Bovinos Identificados En La Prueba De Ifn-γ Usandounclassified

“…Estudios previos indican que los bovinos con mayor número de lesiones producen más óxido nítrico, TNF-α e IFN-γ, por lo que es posible plantear la hipótesis de que los animales identificados en este estudio no pudieron controlar la infección por M. bovis a pesar de la fuerte respuesta inmunitaria proinflamatoria que en este caso, solo aumentó la patología de la enfermedad (21) . Por otra parte, estudios previos han mostrado que los macrófagos de bovinos resistentes producen más óxido nítrico que los macrófagos de bovinos susceptibles (19) .…”

Section: Cuadro 1 Bovinos Identificados En La Prueba De Ifn-γ Usandounclassified

Mycobacterium bovis induce una respuesta inmune celular heterogénea en bovinos infectados naturalmente

López¹,

Flores-Villalva

Pabello³

2017

Rev. Mex. Cienc. Pecu.

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la hipótesis de que las limitaciones en el funcionamiento de los macrófagos y los linfocitos T desempeñan un papel muy importante en la diseminación de la enfermedad.Palabras clave: IFN-γ, Macrófagos, Mycobacterium bovis, PPDB, Coctel proteico. Abstract:Macrophages are considered one of the main immune mechanisms of resistance to mycobacterial infections. In addition, experimental and clinical data indicate that IFN-γ is essential for host defence against M. tuberculosis. The aim of this study was to identify macrophage capacity on controlling Mycobacterium bovis infection and lymphocyte IFN-γ production from cattle naturally exposed to M. bovis. Peripheral blood monocyte-derive macrophages from naturally infected cattle were evaluated for their ability to control the intracellular growth of M. bovis BCG. Besides, whole blood cells were stimulated with PPDB and ESAT-6+CFP-10 protein cocktail. Results show a disparity in host susceptibility to Mycobacterium bovis. A M. bovis BCG survival rate of 80 to 150 % in macrophages from naturally infected cattle was observed. While the IFN-γ production towards Purified protein derivative bovine (PPDB) and the protein cocktail was 2.5 and 2.0 optic density index, respectively. Though our data do not support a correlation between the host susceptibility to M. bovis and the production of IFN-γ in naturally infected cattle, the hypothesis that the limitations in the functioning of macrophages and T lymphocytes play a very important role in the spread of the disease cannot be discarded.

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

Cited by 32 publications

References 38 publications

Specific Recognition of Mycobacterial Protein and Peptide Antigens by γδ T Cell Subsets following Infection with VirulentMycobacterium bovis

Specific Recognition of Mycobacterial Protein and Peptide Antigens by γδ T Cell Subsets following Infection with VirulentMycobacterium bovis

Optimization of a Whole-Blood Gamma Interferon Assay for Detection of Mycobacterium bovis -Infected Cattle

Mycobacterium bovis induce una respuesta inmune celular heterogénea en bovinos infectados naturalmente

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