NKp46/NCR1 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.This product is manufactured by and sold under license from OriGene Technologies and its use is limited solely for research purposes.

NKp46 (also known as natural cytotoxicity receptor 1, NCR1), is a ~46 kDa Type 1 transmembrane protein and a member of the immunoglobulin superfamily (1). NKp46 is highly conserved in mammals and can be characterized by two extracellular C2-type Ig-like domains followed by a stalk region (1). NKp46 is expressed by activated and resting natural killer (NK) cells, innate lymphoid cell (ILC) 1, and a small population of T lymphocytes (1). NKp46, along with NKp30 and NKp44, are activating receptors that have been collectively termed the natural cytotoxicity receptors (NCRs) (2). The NKp46 and NKp30 receptors play a major role in triggering some of the key lytic activities of NK cells (2). NKp46 is the only cytotoxicity receptor with a mouse orthologue, known as mNKp46 (2).

NKp46 plays a critical role in the NK-cell lysis of allogenic, autologous, and xenogeneic cells (3). Activation of NKp46 signaling is mediated via its association with immunoreceptor tyrosine-based activation motif (ITAM) bearing molecules CD3zeta and FcRgamma (3). Upon receptor-engagement NK-cell cytotoxic activity and cytokine release occurs (3). This NKp46 signaling can be blocked with anti-NKp46 monoclonal antibody mediated cross-linking which causes calcium release and interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) secretion (3). NKp46 recognizes multiple ligands presented on tumor cells or pathogens and infected cells including influenza virus hemagglutinin and the parainfluenza virus hemagglutinin neuraminidase (2). Additional ligands for the NKp46 receptor include heparan sulfate (HS) glycosaminoglycans (GAGs), hemagglutinin of human vaccina virus, and vimentin expressed on cells infected with Mycobacterium tuberculosis (3).

NKp46 and its NCR counterparts have shown to be successful targets for cancer immunotherapy. More specifically, human and mouse NKp46 has shown promise in virotherapy due to its ability to stimulate NK cell activation by binding to reovirus sigma1 protein in a sialic acid-dependent manner (3). Another approach for NKp46 in cancer immunotherapy is through the use of NCR based chimeric antigen receptors (CARs). In this therapy, NCR1 supplied human T cells with anti-tumor properties via cytokine secretion, upregulation of activation markers, improved expansion, and cytotoxicity in both mouse and in vitro models (3).

References

1. Barrow, A. D., Martin, C. J., & Colonna, M. (2019). The Natural Cytotoxicity Receptors in Health and Disease. Frontiers in immunology, 10, 909. https://doi.org/10.3389/fimmu.2019.00909

2. Mandelboim, O., & Porgador, A. (2001). NKp46. The international journal of biochemistry & cell biology, 33(12), 1147-1150. https://doi.org/10.1016/S1357-2725(01)00078-4

3. Barrow, A. D., Martin, C. J., & Colonna, M. (2019). The Natural Cytotoxicity Receptors in Health and Disease. Frontiers in immunology, 10, 909. https://doi.org/10.3389/fimmu.2019.00909

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBP2-09751

• NKp46/NCR1 Antibodies
• NKp46/NCR1 Lysates
• NKp46/NCR1 Proteins and Enzymes