Generation and characterization of monoclonal antibodies to equine NKp46.

Veterinary Immunology and Immunopathology
Leela E NoronhaDouglas F Antczak

Abstract

The immunoreceptor NKp46 is considered to be the most consistent marker of NK cells across mammalian species. Here, we use a recombinant NKp46 protein to generate a panel of monoclonal antibodies that recognize equine NKp46. The extracellular region of equine NKp46 was expressed with equine IL-4 as a recombinant fusion protein (rIL-4/NKp46) and used as an immunogen to generate mouse monoclonal antibodies (mAbs). MAbs were first screened by ELISA for an ability to recognize NKp46, but not IL-4, or the structurally related immunoreceptor CD16. Nine mAbs were selected and were shown to recognize full-length NKp46 expressed on the surface of transfected CHO cells as a GFP fusion protein. The mAbs recognized a population of lymphocytes by flow cytometric analysis that was morphologically similar to NKp46+ cells in humans and cattle. In a study using nine horses, representative mAb 4F2 labeled 0.8-2.1% PBL with a mean fluorescence intensity consistent with gene expression data. MAb 4F2+ PBL were enriched by magnetic cell sorting and were found to express higher levels of NKP46 mRNA than 4F2- cells by quantitative RT-PCR. CD3-depleted PBL from five horses contained a higher percentage of 4F2+ cells than unsorted PBL. Using ELISA, we d...Continue Reading

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Mar 20, 2012·Veterinary Immunology and Immunopathology·Leela E NoronhaDouglas F Antczak

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Citations

Mar 5, 2013·Veterinary Research·Kerstin H MairWilhelm Gerner
Sep 2, 2014·Veterinary Immunology and Immunopathology·Gillian A PerkinsBettina Wagner
Oct 22, 2013·Veterinary Immunology and Immunopathology·C L SchnabelJ-M V Cavalleri
Dec 2, 2014·Veterinary Research Communications·Christine Grøndahl-RosadoAnne K Storset
Mar 19, 2014·The Journal of Immunology : Official Journal of the American Association of Immunologists·Timothy K ConnelleyW Ivan Morrison
Aug 1, 2012·Reproduction in Domestic Animals = Zuchthygiene·D F Antczak

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