tumor cell lysis

1
T Cell-Mediated Tumor Cell Lysis Assay

• Reference
• https//en.wikipedia.org/wiki/Chimeric_antigen_rec
  eptor_T_cell
• https//www.creative-biolabs.com/car-t/t-cell-medi
  ated-tumor-cell-lysis-assay.htm

2
Background

• Chimeric antigen receptors (CARs, also known as
  chimeric immunoreceptors, chimeric T cell
  receptors or artificial T cell receptors) are
  receptor proteins that have been engineered to
  give T cells the new ability to target a specific
  protein. The receptors are chimeric because they
  combine both antigen-binding and T-cell
  activating functions into a single receptor.
• CAR-T cell therapy uses T cells engineered with
  CARs for cancer therapy. The premise of CAR-T
  immunotherapy is to modify T cells to recognize
  cancer cells in order to more effectively target
  and destroy them. Scientists harvest T cells from
  people, genetically alter them, then infuse the
  resulting CAR-T cells into patients to attack
  their tumors.

3

• Anti-tumor activity in mice is induced only when
  both the universal CAR T cells plus the correct
  antigen-specific adaptor molecules are present.
  Anti-tumor activity and toxicity can be
  controlled by adjusting the administered adaptor
  molecule dosing. Treatment of antigenically
  heterogeneous tumors can be achieved by
  administration of a mixture of the desired
  antigen-specific adaptors. Thus, several
  challenges of current CAR T cell therapies, such
  as
• the inability to control the rate of cytokine
  release and tumor lysis
• the absence of an off switch that can terminate
  cytotoxic activity when tumor eradication is
  complete
• a requirement to generate a different CAR T cell
  for each unique tumor antigen
• may be solved or mitigated using this approach.

4
Two Assays for T Cell-Mediated Tumor Cell Lysis

• The target tumor cells can be incubated with the
  radiolabel. The specific radiolabel is taken up
  and reversibly binds to cytosolic proteins. When
  the target cells are recognized by CAR-T
  lymphocytes, the target cells are killed and the
  radiolabel is released.

5
Two Assays for T Cell-Mediated Tumor Cell Lysis

• The target tumor cells are labeled with specific
  fluorescent cytoplasmic reagent, the live cells
  can be imaged and monitored at real-time level to
  measure the tumor cell lysis. The reagent is safe
  for cells at optimized concentration, and
  fluorescence measurements can be accurately
  performed for short-term experiments, as the
  reagent will be diluted with cell proliferation.
  In addition, the ratio of target tumor cells and
  CAR-T cells can be optimized in the assay.

6
(No Transcript)
7
Thanks

• Reference
• https//en.wikipedia.org/wiki/Chimeric_antigen_rec
  eptor_T_cell
• https//www.creative-biolabs.com/car-t/t-cell-medi
  ated-tumor-cell-lysis-assay.htm