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Enhancement or Inhibition of T Cell Response Assays

As a leading service provider in the field of antibody discovery and immunotherapy, Creative Biolabs now presents one-stop CAR-T therapy development service which has become a revolution in the area of cancer therapy. To verify the superior efficiency of CAR-engineered T cells, we have a serial assay to detect the enhancement or inhibition phenotype of T cell response.

1. 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. The efficiencies of T cell response include T cell proliferation, differentiation, functional cytokines secretion, and cytotoxic T lymphocyte function etc. To validate the T cell response, Creative Biolabs now provides the services of the following assays but not limited to these.

2. T cell response assays

Creative Biolabs offers various assays to enhance or inhibit T cell response by using different cytokines, antibodies, and co-stimulators. Of note, we are capable of performing versatile assays to evaluate the T cell response.

➢ 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.

T cell response - proliferation and differentiation

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

➢ 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. Creative Biolabs provides all kinds of fluorescent antibodies to detect the surface biomarkers or transcription factors to identify different T cell subsets via FACS. For example, we have the antibodies to identify Treg - CD4, FOXP3, and CD25; also we have the antibodies to identify the key transcription factors of Th1 - STAT4, T-bet and Th2 - STAT6, GATA3.

Schematic representation of various T cell subsets

Figure 2: Schematic representation of various T cell subsets (O'Shea and Paul. 2010)

➢ 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. Thus to detect the cytokines secretion is an important indicator to understand the efficiency and function of T cell response. Creative Biolabs can provide the assays: ELISPOT, ELISA, profile array, FACS, Immunofluorescence, ICS etc. We aim to provide relevant antibodies with high quality, high sensitivity, broad dynamic range, high affinity and specificity.

Different functional cytokines

Figure 3: Different functional cytokines (Butterfield, et.al. 2017)

➢ 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. Creative Biolabs can provide CTL assay through radioactive chromium-release, flow cytometric monitoring of target cell lysis, effector cell frequency & activity, or quantification of CTL precursors etc.

CTL-mediated cytotoxicity

Figure 4. CTL-mediated cytotoxicity (Andersen, et.al. 2006)

➢ T Cell Infiltration Assay

Infiltration, accumulation, and survival of chimeric antigen receptor T (CAR-T) cells in solid tumors are crucial for tumor clearance. Creative Biolabs can provide customers with T cell infiltration/migration assay to ensure the functional T cells are located in tumor microenvironment to display humoral or cellular immunity both in vitro and in vivo.

CAR-T infiltrated tumor microenvironment

Figure 5. CAR-T infiltrated tumor microenvironment (Newick, et.al. 2016)

Highlight Features of Our Service

  • Ph.D. level scientists and skillful technicians
  • Rich experience and advanced platforms
  • Time- and labor-saving
  • Superior quality
  • High-quality after-sales service

With years of extensive experience in the field of immunotherapy, Creative Biolabs has won a good reputation and established cooperation with numerous worldwide customers. We promise timely troubleshooting to guarantee the highest success rate of every project. We are dedicated to facilitating your projects and pushing the progress towards clinical trials through sharing our technologies, platforms, and resources. We can provide the above T cell response detection assays but not limited to those. Any customized consultant is welcomed. Please don't hesitate to contact us for more details and our team will get back to you as soon as possible.

References

  1. Butterfield LH, et al. (2017). Sequence of T cell activation. Cancer Immunotherapy Principles and Practice. p89.
  2. O'Shea JJ and Paul WE. (2010) Mechanisms underlying lineage commitment and plasticity of helper CD4+ T cells. Science. 327(5969):1098-102.
  3. Andersen MH, et.al. (2006) Cytotoxic T Cells Journal of Investigative Dermatology. 126, 32–41.
  4. Newick K, et.al. (2016) Chimeric antigen receptor T-cell therapy for solid tumor. Annu Rev Med.

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