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",

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journal = "Developmental and Comparative Immunology",

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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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