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Originally published online as doi:10.1189/jlb.0408255 on August 15, 2008

Published online before print August 15, 2008
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(Journal of Leukocyte Biology. 2008;84:1501-1510.)
© 2008 by Society for Leukocyte Biology

CD5-low expression lymphocytes in canine peripheral blood show characteristics of natural killer cells

Yi-Chun Huang*, Shao-Wen Hung{dagger}, Tong-Rong Jan*, Kuang-Wen Liao{ddagger}, Chiung-Hsiang Cheng*, Yu-Shan Wang* and Rea-Min Chu*,1

* Animal Cancer Research Center, Department of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, Taipei, Taiwan;
{dagger} Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan; and
{ddagger} Department of Biological Sciences and Technology, College of Life Sciences, Hsin-Chu, Taiwan

1 Correspondence: Animal Cancer Research Center, Department of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei 106, Taiwan. E-mail: redman{at}ntu.edu.tw

NK cell markers and receptors have been discovered in many mammalian species, such as humans, mice, rats, pigs, and cows. However, there is still a lack of information concerning NK cell markers or receptors in canines. We have discovered that canine CD5-low density (CD5lo) cells in PBL are closely associated with NK cell characteristics. CD5lo cells comprised 14.9 ± 6.68% of the total PBL. A high proportion of the CD5lo cell population expressed CD3 (96.6%), CD8{alpha} (77.7%), CD8β (53%), {alpha}/β TCR (83%), and CD11/18 (80%), but the expression of {gamma}/{delta} TCR (6.5%), CD4 (10.6%), and CD21 (2.4%) was low. CD5lo cells were larger than CD5-high density (CD5hi) cells. Light and electron microscopy revealed numerous large cytoplasmic granules in CD5lo cells, especially after IL-2 stimulation, which was in contrast to CD5hi, in which intracytoplasmic granules were not frequently seen. After IL-2 stimulation, CD5lo cells had significantly stronger NK cytotoxicity than CD5hi cells. CD5lo cells had much higher mRNA levels for NKG2D, CD16, CD94, CD160, perforin, and granzyme than CD5hi. Following IL-2 stimulation, CD5lo cells had significantly higher mRNA levels of NKp30, NKp44, CD16, and CD94 than CD5hi cells. In addition, IL-2-stimulated, CD5lo-depleted PBL showed a loss of NK cytotoxicity. CD5lo cells also showed significantly lower antigen-specific cytotoxic T cell activity as compared with CD5hi cells. Taken together, the CD5lo subset in canine PBL is closely related to canine NK cells, and CD5lo can be used as a phenotypic marker for an IL-2-dependent canine NK cell enrichment.

Key Words: cytotoxicity • LDH-releasing test • T cell markers