Journal of Leukocyte Biology eBioscience full spectrum cell analysis
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Originally published online as doi:10.1189/jlb.0505239 on November 7, 2005

Published online before print November 7, 2005
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(Journal of Leukocyte Biology. 2006;79:71-79.)
© 2006 by Society for Leukocyte Biology

Bovine natural killer cells acquire cytotoxic/effector activity following activation with IL-12/15 and reduce Mycobacterium bovis BCG in infected macrophages

Janice J. Endsley*,1, Mark A. Endsley{dagger} and D. Mark Estes*,{ddagger},{dagger}

Departments of
* Pediatrics and Sealy Center for Vaccine Development and
{dagger} Microbiology and Immunology, University of Texas Medical Branch, Galveston; and
{ddagger} Departments of Veterinary Pathobiology and Molecular Microbiology and Immunology, University of Missouri-Columbia

1 Correspondence: 2.120 Children’s Hospital, University of Texas Medical Branch, Galveston, TX 77555. E-mail: jjendsle{at}utmb.edu

Bovine natural killer (NK) cells were recently identified by positive selection of a NK cell-activating receptor p46 (NKp46)+ CD3– lymphocyte population, which expresses CD25 and CD8 and lyses tumor cell lines following stimulation with recombinant interleukin-2. In the current work, we characterize the cytotoxic/effector potential of a CD3–CD8–CD11b– population isolated through negative selection of bovine peripheral blood leukocytes. This population is CD25loCD62hi when isolated and becomes CD25hiCD62Llo following cytokine stimulation. Activated bovine NK cells increase expression of granulysin, interferon-{gamma}, and perforin and have cytotoxic activity against human tumor cells and Mycobacterium bovis bacillus Calmette-Guerin-infected alveolar and monocyte-derived macrophages. Expression of a bovine homologue of the CD56 neural adhesion molecule expressed by human NK cells was detected in mRNA from brain tissue but was not detected in peripheral blood mononuclear cells or purified NK cell mRNA. Analysis of mRNA from nonstimulated peripheral blood NK cells demonstrates the constitutive expression of homologues of human NK receptors NKp46, CD244, and CD94 and the granule proteins granulysin and perforin. Phorbol ester-stimulated CD8+ T cells also expressed CD244 and CD94, and CD4+ T cells expressed CD94. These NK cell receptors bearing T lymphocytes may represent memory subsets characterized in humans. The results of these studies demonstrate that bovine NK cells may play an important role in the innate immune responses of cattle.

Key Words: cytotoxic granule proteins • natural killer receptors • tuberculosis






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