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Assessment of Human Natural Killer Cell Events Driven by FcγRIIIa Engagement in the Presence of Therapeutic Antibodies
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Assessment of Human Natural Killer Cell Events Driven by FcγRIIIa Engagement in the Presence of Therapeutic Antibodies

Assessment of Human Natural Killer Cell Events Driven by FcγRIIIa Engagement in the Presence of Therapeutic Antibodies

DOI:
10.3791/61144-v

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09:54 min

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May 22, 2020

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, , , , , ,
1Department of Research and Development,iQ Biosciences, 2Department of Cellular Products,iQ Biosciences

Chapters

  • 00:05Introduction
  • 00:27Antibody-Mediated Activation of NK Cells
  • 03:12Interrogation of Signaling Molecules by Western Blot Analysis
  • 03:47Isolation of mRNA and Preparation for Chemokine/Cytokine Assessment
  • 04:39Cytoskeletal Rearrangement Assessment
  • 05:57Assessing Degranulation Using CD107a Surface Staining
  • 07:10Results: Downstream Applications of NK Cell Activation Assay
  • 09:20Conclusion

Summary

Automatic Translation

Automatic Translation

A protocol for studying FcγRIIIa-driven events by therapeutic antibodies in human natural killer cells is described here. This artificial stimulation platform permits the interrogation of downstream effector functions such as degranulation, chemokine/cytokine production, and signaling pathways mediated by the FcγRIIIa and Fc portions of antibodies involved in binding.

Tags

Keywords: Fc Gamma RIIIANatural Killer CellsTherapeutic AntibodiesCellular Molecular MechanismActivationCytoskeletal RearrangementWestern Blot AnalysisRIPA BufferProtein Separation

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