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CD8+ T cells are a vital branch of the adaptive immune response. They are necessary for an effective antiviral and anticancer immunity, which is mediated through their cytotoxic effector functions. CD8+ T cell activation, proliferation, and effector functions are dependent on a complex network of Ca2+ signalling pathways involving both Ca2+ release and Ca2+ entry. Dysregulation of intracellular Ca2+ signalling pathways has been linked to immunodeficiencies including severe combined immunodeficiency syndrome (SCID), highlighting the need to study both Ca2+ dynamics and cytotoxic effector function in patient-derived CD8+ T cells. The following protocol describes a workflow for the isolation of primary human CD8+ T cells and their activation using bispecific T cell engagers (BiTEs). This allows for a simultaneous assessment of both Ca2+ dynamics and cytotoxicity through a target cell line presenting an epitope recognized by the BiTE.

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Imaging BiTE-Mediated Activation of Primary Human CD8+ T Cells

  • 实验方案

  • 发表日期：

    2025年

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  • Kai J. Winterberg,
  • Constantin Möller,
  • Lea I. Schwarze,
  • Sophia Cichutek,
  • Julia Hambach,
  • Andreas H. Guse,
  • Boris Fehse,
  • Björn-Philipp Diercks,
  • Kristoffer Riecken

摘要

CD8+ T cells are a vital branch of the adaptive immune response. They are necessary for an effective antiviral and anticancer immunity, which is mediated through their cytotoxic effector functions. CD8+ T cell activation, proliferation, and effector functions are dependent on a complex network of Ca2+ signalling pathways involving both Ca2+ release and Ca2+ entry. Dysregulation of intracellular Ca2+ signalling pathways has been linked to immunodeficiencies including severe combined immunodeficiency syndrome (SCID), highlighting the need to study both Ca2+ dynamics and cytotoxic effector function in patient-derived CD8+ T cells. The following protocol describes a workflow for the isolation of primary human CD8+ T cells and their activation using bispecific T cell engagers (BiTEs). This allows for a simultaneous assessment of both Ca2+ dynamics and cytotoxicity through a target cell line presenting an epitope recognized by the BiTE.