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

C.A. Jansen; P.M. van de Haar; D.A. van Haarlem; Peter van Kooten; Sjaak de Wit; W. van Eden; B.C. Viertlböck; T.W. Göbel; L. Vervelde

doi:10.1016/j.dci.2010.02.009

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

C.A. Jansen
, P.M. van de Haar
, D.A. van Haarlem
, Peter van Kooten
, Sjaak de Wit
, W. van Eden
, B.C. Viertlböck
, T.W. Göbel
, L. Vervelde

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

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−CD8α+ 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.

Original language	English
Pages (from-to)	759-767
Number of pages	8
Journal	Developmental and Comparative Immunology
Volume	34
Issue number	7
DOIs	https://doi.org/10.1016/j.dci.2010.02.009
Publication status	Published - 2010

Keywords

NK-cells
Chickens
Markers
CD107

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title = "Identification of new populations of chicken natural killer (NK) cells",

abstract = "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−CD8α+ 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.",

keywords = "NK-cells, Chickens, Markers, CD107",

author = "C.A. Jansen and \{van de Haar\}, P.M. and \{van Haarlem\}, D.A. and \{van Kooten\}, Peter and \{de Wit\}, Sjaak and \{van Eden\}, W. and B.C. Viertlb{\"o}ck and T.W. G{\"o}bel and L. Vervelde",

year = "2010",

doi = "10.1016/j.dci.2010.02.009",

language = "English",

volume = "34",

pages = "759--767",

journal = "Developmental and Comparative Immunology",

issn = "0145-305X",

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T1 - Identification of new populations of chicken natural killer (NK) cells

AU - Jansen, C.A.

AU - van de Haar, P.M.

AU - van Haarlem, D.A.

AU - van Kooten, Peter

AU - de Wit, Sjaak

AU - van Eden, W.

AU - Viertlböck, B.C.

AU - Göbel, T.W.

AU - Vervelde, L.

PY - 2010

Y1 - 2010

N2 - 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−CD8α+ 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.

AB - 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−CD8α+ 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.

KW - NK-cells

KW - Chickens

KW - Markers

KW - CD107

U2 - 10.1016/j.dci.2010.02.009

DO - 10.1016/j.dci.2010.02.009

M3 - Article

SN - 0145-305X

VL - 34

SP - 759

EP - 767

JO - Developmental and Comparative Immunology

JF - Developmental and Comparative Immunology

IS - 7

ER -

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

Abstract

Keywords

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