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Rat NKp46 activates natural killer cell cytotoxicity and is associated with FcɛRIγ and CD3ζ
Author(s) -
Westgaard Ingunn H.,
Berg Siri F.,
Vaage John T.,
Wang Lawrence L.,
Yokoyama Wayne M.,
Dissen Erik,
Fossum Sigbjørn
Publication year - 2004
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1189/jlb.0903428
Subject(s) - biology , cd16 , natural killer cell , lymphokine activated killer cell , microbiology and biotechnology , effector , monoclonal antibody , flow cytometry , cytotoxicity , interleukin 21 , cd3 , antibody , t cell , immunology , antigen , immune system , in vitro , cd8 , biochemistry
NKp46 has been identified in the human, rat, mouse, monkey, and cattle. We have generated a monoclonal antibody, WEN23, against rat NKp46. By flow cytometry, NKp46 is expressed by all natural killer (NK) cells but not by T cells, B cells, granulocytes, monocytes, dendritic cells, or macrophages. Thus, NKp46/WEN23 is the first NK cell‐specific marker in the rat. In a redirected lysis assay, preincubation of the effector cells with WEN23 augmented lysis of the Fc receptor (FcR) + murine tumor target cells, indicating that NKp46 is an activating NK cell receptor. Moreover, preincubation of the effector cells with WEN23 F(ab′) 2 fragments reduced killing of target cells, confirming the activating function of NKp46 and indicating that the mouse tumor target cells express a ligand for rat NKp46. Lysis of FcR − mouse and human tumor target cells was reduced after incubation of effector cells with WEN23, suggesting that rat NKp46 recognizes a ligand that is conserved between primates and rodents. By Western blot and immunoprecipitation using WEN23, NKp46 is expressed as a monomer of ∼47 kDa in interleukin‐2‐activated NK cells. The immunoreceptor tyrosine‐based activation motif bearing adaptor proteins CD3ζ and the γ chain of FcRI for IgE (FcɛRIγ) with NKp46 from lysates of NK cells, indicating that rat NKp46 activates NK cell cytotoxicity by similar pathways as CD16.