NK Cell Killing

NK Cell Killing Application
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Natural Killer (NK) cells are granular lymphocytes involved in the innate immune response to viruses and tumors. NK cells have gathered significant attention in the field of cancer immunotherapy as a potential allogeneic “off-the-shelf” therapeutic because they can: 

  • >> Kill tumor cells without antigen expression 
    >> Kill multiple cells, potentially reducing dose 
    >> Reduce risk of cytokine release syndrome 
    >> Don’t have the same HLA compatibility issues as T cells 

Activation and Inhibition of Natural Killer Cells

Important to the innate immune system, natural killer cells (or large granular lymphocytes) kill diseased cells. They express a range of activating and inhibitory receptors that regulate antitumor activity.

Activating receptors​

  • NKG2D​
  • CD2​
  • NCR​
  • CD16​

Inhibitory receptors​

  • KIRs​
  • CD94​
  • ILT/LIR
In vitro Natural Killer Cell (NK cells) assays to assess immunopotency for killing disease cells.  Both activating and inhibitory receptors can regulate antitumor activity

NK Cell Killing Assays

Assess NK-mediated killing with cancer-targeting antibodies
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Purpose: To demonstrate antibody-dependent cellular cytotoxicity using NK cells. NK cells kill cells that have antibodies bound to their surface, making them useful for assessing antibody-based therapies. 

Trastuzumab promotes antibody-dependent NK cell killing of A172 target cells.
Trastuzumab promotes antibody-dependent NK cell killing of A172 target cells.
Trastuzumab promotes antibody-dependent NK cell killing of A172 target cells.
Trastuzumab promotes antibody-dependent NK cell killing of A172 target cells.

Cancer cells were treated with trastuzumab, a HER2-targeting monoclonal antibody, and then co-cultured with NK cells. NK-mediated killing of target cells was assessed using the Maestro Z platform. 

Result: NK cell-mediated killing was greatly enhanced in the presence of trastuzumab. 

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Measure potency of CAR NK cells
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Purpose: To demonstrate enhanced NK-mediated targeting and cytotoxicity. Like CAR T cell development, antigen selection plays a crucial role in therapeutic development of CAR NKs.

Cytolysis of HER2-expressing cancer cells by NK cells possessing HER2-targeting chimeric antigen receptor (CAR).
Cytolysis of HER2-expressing cancer cells by NK cells possessing HER2-targeting chimeric antigen receptor (CAR).
Cytolysis of HER2-expressing cancer cells by NK cells possessing HER2-targeting chimeric antigen receptor (CAR).
Cytolysis of HER2-expressing cancer cells by NK cells possessing HER2-targeting chimeric antigen receptor (CAR).

NK cells transduced with chimeric antigen receptor (CAR) constructs targeting HER2 or CD19 were added to HER2-expressing cancer cells. CAR NK-mediated cytotoxicity was measured using the Maestro Z platform.  

Result: HER2-targeted CAR NK cells exhibited higher potency than CD19-targeted or non-transduced NK cells. The difference in cytolysis between non-transduced and HER2 CAR NK cells demonstrates that antigen recognition and specificity is an important determinant for CAR NK-cell potency. 

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