T cell infiltration

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Enhancement or Inhibition of T Cell Response
Assays

• Reference https//www.creative-biolabs.com/car-t/
  enhancement-or-inhibition-of-t-cell-response-assay
  s.htm

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Background

• Adoptive therapy with chimeric antigen receptor
  (CAR) T cells is an innovative approach to
  program the immune system to attack malignant
  tumor. It uses genetically engineered immune T
  cells to recognize specific proteins on tumor
  cells through corresponding humoral immunity and
  cellular immunity. Except for safety, efficiency
  of CAR-T cell is quite crucial important to
  eliminate various kinds of tumors.

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Proliferation Assay

• T cell activation induces T cell proliferation,
  with the goal of clonally selecting and expanding
  antigen-specific T cells which can induce an
  effective, long-lasting adaptive immunity. To
  detect the extent of proliferation, we have CFSE
  assay and 3H-thymidine incorporation assay.
• Carboxyfluorescein succinimidyl ester (CFSE) is a
  fluorescent cell staining dye which can be used
  to monitor lymphocyte proliferation, both in
  vitro and in vivo. The thymidine incorporation
  assay utilizes a strategy wherein a radioactive
  nucleoside, 3H-thymidine, is incorporated into
  new strands of chromosomal DNA during mitotic
  cell division. Both assays can be applied to
  determine the extent of T cell proliferation that
  has occurred in response to specific T cell
  stimulation.

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Figure 1 T cell response - proliferation and
differentiation (Butterfield, et.al. 2017)
5
Differentiation Assay

• T cell activation induces T cell differentiation
  into different subsets including T helper cell
  (Th1, Th2, Th17 etc.), CD8 T cell, Treg cell
  etc. once stimulated by specific antigens
  presented by APC. The differentiation is
  characterized by expression of various CD surface
  molecules. These different subsets have important
  implications for tumor treatment.

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Figure 2 Schematic representation of various T
cell subsets (O'Shea and Paul. 2010)
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Functional Cytokines Producing Assay

• The recognition between TCR and specific antigens
  presented by APC can induce T cell activation and
  a series of functional cytokines secretion.
  Cytokines are small cell-signaling protein
  molecules involved in cellular communication,
  immune response, inflammation, hematopoiesis and
  they are crucial for determining anti-tumor
  responses.

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Figure 3 Different functional cytokines
(Butterfield, et.al. 2017)
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Cytotoxic T Lymphocyte Function Assay

• Evaluation of cell-mediated immunity is
  significant to understand the immunotherapy.
  Cytotoxic T lymphocyte response is recognized as
  perhaps the most relevant functional measure that
  reflects cell-mediated acquired immune defense
  against cancer. Therefore, CTL assay is an
  excellent candidate for evaluation of the
  cytolytic activities.

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Figure 4. CTL-mediated cytotoxicity (Andersen,
et.al. 2006)
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T Cell Infiltration Assay

• Infiltration, accumulation, and survival of
  chimeric antigen receptor T (CAR-T) cells in
  solid tumors are crucial for tumor clearance.
• T cell infiltration assays ensure the functional
  T cells are located in tumor microenvironment to
  display humoral or cellular immunity both in
  vitro and in vivo.

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Figure 5. CAR-T infiltrated tumor
microenvironment (Newick, et.al. 2016)
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Thanks
Reference https//www.creative-biolabs.com/car-t/
enhancement-or-inhibition-of-t-cell-response-assay
s.htm