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International Immunology Advance Access originally published online on August 16, 2004
International Immunology 2004 16(10):1459-1466; doi:10.1093/intimm/dxh147
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© 2004 The Japanese Society for Immunology

Isolation of a novel KIR2DL3-specific mAb: comparative analysis of the surface distribution and function of KIR2DL2, KIR2DL3 and KIR2DS2

Massimo Vitale1, Simona Carlomagno2, Michela Falco3, Daniela Pende1, Elisa Romeo3, Paola Rivera2, Mariella Della Chiesa2, Domenico Mavilio4 and Alessandro Moretta2,5

1 Istituto Nazionale per la Ricerca sul Cancro, L.go R. Benzi 10, 16132 Genova, Italy
2 Dipartimento di Medicina Sperimentale, Università degli Studi di Genova, Via L.B. Alberti 2, 16132 Genova
3 Istituto Giannina Gaslini, L.go G. Gaslini 5, 16148 Genova, Italy
4 Laboratory of Immunoregulation, National Institute of Allergy and Infectious Disease, NIH, Bethesda, MD 20892, USA
5 Centro di Eccellenza per le Ricerche Biomediche, Università degli Studi di Genova, V.le Benedetto XV, 16132 Genova, Italy

Correspondence to: A. Moretta; E-mail: alemoret{at}unige.it

In recent years an increasing number of sequences coding for new KIRs have been described. However, the limited availability of mAbs with unique KIR specificities has hindered an exhaustive assessment of their actual function, HLA-specificity, expression at the cell surface and distribution in different cell populations. In this study we report the generation of a novel mAb (ECM41) specific for KIR2DL3 molecules. By the use of cell transfectants expressing one or other KIR we show that this reagent allows discrimination of KIR2DL3 from other GL183 mAb-reactive molecules such as KIR2DL2 and KIR2DS2. Moreover we show that this novel mAb can be used to assess the surface expression and distribution of KIR2DL3 in different polyclonal NK populations and in NK cell clones. Along this line, we were able to analyze the HLA class I specificity of NK clones expressing either KIR2DL3 or KIR2DL2, two inhibitory receptors that were so far serologically undistinguishable. Finally, the combined use of GL183 and ECM41 mAbs in redirected killing assays allowed us to investigate the functional outcome of the simultaneous engagement of KIR2DL3 and KIR2DS2 in NK cell clones co-expressing KIRs that display opposite (inhibitory vs activating) function.

Keywords: HLA-specific receptors, immunoglobulin superfamily, KIR, NK cells

Transmitting editor: E. Vivier


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