Identification of new populations of chicken natural killer (NK) cells.
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.
Jansen
Christine A
CA
Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, Utrecht, The Netherlands. c.a.jansen@uu.nl
van de Haar
Peter M
PM
van Haarlem
Daphne
D
van Kooten
Peter
P
de Wit
Sjaak
S
van Eden
Willem
W
Viertlböck
Birgit C
BC
Göbel
Thomas W
TW
Vervelde
Lonneke
L
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002645:Chickens / Q000276:immunology*; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D005260:Female; D005434:Flow Cytometry / Q000662:veterinary; D007694:Killer Cells, Natural / Q000276:immunology*; D052118:Lysosomal-Associated Membrane Protein 1 / Q000276:immunology; D013047:Specific Pathogen-Free Organisms; D018709:Statistics, Nonparametric
D000911:Antibodies, Monoclonal; D052118:Lysosomal-Associated Membrane Protein 1