Identification of new populations of chicken natural killer (NK) cells

• DOI: 10.1016/j.dci.2010.02.009
• Publication Date: 2010-03-05T09:41:17Z
• Authors: Christine A. Jansen; Peter M. van de Haar; Daphne van Haarlem; Peter van Kooten; Sjaak de Wit; Willem van Eden; Birgit C. Viertlböck; Thomas W. Göbel; Lonneke Vervelde
• Keywords: Lymphokine-activated killer cell; Janus kinase 3

Natural killer (NK) cell activity is conserved throughout vertebrate development, but characterization of non-mammalian NK-cells has been hampered by the absence of specific mAbs for these cells. Monoclonal antibodies were generated against in vitro IL-2 expanded sorted CD3-CD8alpha+ peripheral blood lymphocytes, previously described to contain chicken NK-cells. Screening of embryonic and adult splenocytes with hybridoma supernatants resulted in five candidate NK markers. Activation of chicken NK-cells with PMA/Ionomycin or with the NK target cell-line LSCC-RP9 resulted in increased expression of CD107 (LAMP-1) and a newly developed flow cytometry based cytotoxicity assay showed that NK-cells were able to kill target cells. Combining NK markers with functional assays indicated that marker positive cells showed NK-cell function. In conclusion, we generated new monoclonal antibodies and developed two functional assays which will enhance our understanding of the role of NK-cells in healthy and diseased chickens.