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Clinical and Vaccine Immunology, July 2009, p. 1003-1011, Vol. 16, No. 7
1071-412X/09/$08.00+0     doi:10.1128/CVI.00114-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Neutralization of Interleukin-10 from CD14+ Monocytes Enhances Gamma Interferon Production in Peripheral Blood Mononuclear Cells from Mycobacterium avium subsp. paratuberculosis-Infected Goats{triangledown}

Kari R. Lybeck,1* Anne K. Storset,2 and Ingrid Olsen1

Department of Animal Health, National Veterinary Institute, Oslo, Norway,1 Department of Food Safety and Infection Biology, Norwegian School of Veterinary Science, Oslo, Norway2

Received 12 March 2009/ Returned for modification 1 April 2009/ Accepted 27 April 2009

The gamma interferon assay is used to identify Mycobacterium avium subsp. paratuberculosis-infected animals. It has been suggested that regulatory mechanisms could influence the sensitivity of the test when it is performed with cells from cattle and that the neutralization of interleukin-10 (IL-10) in vitro would increase the gamma interferon responses. To investigate the regulatory mechanisms affecting the gamma interferon assay with cells from goats, blood was collected from M. avium subsp. paratuberculosis-infected, M. avium subsp. paratuberculosis-exposed, and noninfected goats. Neutralization of IL-10 by a monoclonal antibody resulted in increased levels of gamma interferon production in M. avium subsp. paratuberculosis purified protein derivative (PPDj)-stimulated samples from both infected and exposed goats. However, the levels of gamma interferon release were also increased in unstimulated cells and in PPDj-stimulated cells from some noninfected animals following neutralization. Depletion of putative regulatory CD25high T cells had no clear effect on the number of gamma-interferon-producing cells. The IL-10-producing cells were identified to be mainly CD14+ major histocompatibility complex class II-positive monocytes in both PPDj-stimulated and control cultures and not regulatory T cells. However, possible regulatory CD4+ CD25+ T cells produced IL-10 in response to concanavalin A stimulation. The numbers of CD4+, CD8+, and CD8+ {gamma}{delta}T-cell receptor-positive cells producing gamma interferon increased following IL-10 neutralization. These results provide insight into the source and the role of IL-10 in gamma interferon assays with cells from goats and suggest that IL-10 from monocytes can regulate both innate and adaptive gamma interferon production from several cell types. Although IL-10 neutralization increased the sensitivity of the gamma interferon assay, the specificity of the test could be compromised.

* Corresponding author. Mailing address: Department of Animal Health, National Veterinary Institute, Pb 750 Sentrum, Oslo 0106, Norway. Phone: 4723216341. Fax: 4723216302. E-mail: kari.lybeck{at}vetinst.no

{triangledown} Published ahead of print on 6 May 2009.

Clinical and Vaccine Immunology, July 2009, p. 1003-1011, Vol. 16, No. 7
1071-412X/09/$08.00+0     doi:10.1128/CVI.00114-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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