Development and characterization of a canine-specific anti-CD94 (KLRD-1) monoclonal antibody

Scott S Graves, Boglarka Gyurkocza, Diane M Stone, Maura H Parker, Kraig Abrams, Christoph Jochum, Susanna Gallo, Marium Saad, Melissa M Johnson, Steven L Rosinski, Rainer Storb

Research output: Contribution to journalArticlepeer-review

Abstract

Natural killer (NK) cells are non-T, non-B lymphocytes are part of the innate immune system and function without prior activation. The human NK cell surface determinant, CD94, plays a critical role in regulation of NK cell activity as a heterodimer with NKG2 subclasses. Canine NK cells are not as well defined as the human and murine equivalents, due in part to the paucity of reagents specific to cell surface markers. Canines possess NK/NKT cells that have similar morphological characteristics to those found in humans, yet little is known about their functional characteristics nor of cell surface expression of CD94. Here, we describe the development and function of a monoclonal antibody (mAb) to canine (ca) CD94. Freshly isolated canine CD94+ cells were CD3+/-, CD8+/-, CD4-, CD21-, CD5low, NKp46+, and were cytotoxic against a canine target cell line. Anti-caCD94 mAb proved useful in enriching NK/NKT cells from PBMC for expansion on CTAC feeder cells in the presence of IL-2 and IL-15. The cultured cells were highly cytolytic with co-expression of NKp46 and reduced expression of CD3. Transmission electron microscopy revealed expanded CD94+ lymphocytes were morphologically large granular lymphocytes with large electron dense granules. Anti-caCD94 (mAb) can serve to enrich NK/NKT cells from dog peripheral blood for ex vivo expansion for HCT and is a potentially valuable reagent for studying NK/NKT regulation in the dog.

Original languageEnglish
Pages (from-to)10-18
Number of pages9
JournalVeterinary Immunology and Immunopathology
Volume211
DOIs
Publication statusPublished - May 2019

Keywords

  • Animals
  • Antibodies, Monoclonal/immunology
  • Antibody Specificity/immunology
  • Cloning, Molecular
  • Dogs/immunology
  • Female
  • Flow Cytometry/veterinary
  • Killer Cells, Natural/immunology
  • Male
  • Mice/immunology
  • NK Cell Lectin-Like Receptor Subfamily D/immunology
  • Natural Killer T-Cells/immunology
  • Polymerase Chain Reaction/veterinary

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