T Cell-based In Vitro Assay - Creative Biolabs

Creative Biolabs has developed a variety of T and B cell-based assays to analyze cancer antigens and identify immunodominant epitopes based on extensive, versatile techniques. The in vitro assay is a well-established and reproducible method for characterizing epitope-induced T cell responses, such as cytotoxicity, cell proliferation, and suppression function. Based on your sample, our experienced scientists can provide proper assays to accelerate your target-mediated immunotherapy development.

Cytokine Release Assay

Cytokine release is a crucial signature of activated T cells and can be used to screen the cancer epitope that can induce immune responses. ELISA assay is typically applied to measure cytokine production quantitatively. With access to various commercial ELISA kits and high-quality antibodies, we provide a comprehensive profile of cytokine secretion activities.

Techniques	Functions	Advantages	Limitations
ELISA	Detect Cytokine Secretion	Sensitive Accurate Widely Used	Laborious Limited Detection Range
Advanced ELISA	Detect Cytokine Secretion	High Throughput Greater Sensitivity Wide Range of Detection Limits Free of Washing Steps	None

The ELISA-related techniques include but are not limited to the following.

CD4+ T cells involved antitumor immunity. (Kim & Cantor, 2014) Fig.1 CD4+ T cells involved in antitumor immunity.¹

Cell Proliferation Assay

Cell proliferation is the phenomenon of the clonal growth of T and B cells resulting from antigen stimulation and is one of the measurement markers of successful cell activation.

Technology	Principle	Correlate Techniques	Advantages	Limitations
CFSE Assay	Reduced CFSE Signal as Cell Division	Flow Cytometry	Simple to Perform Relatively High Throughput Non-radioactivity	May Influence Cell Viability
³H-thymidine Assay	Incorporate in Dividing Cells	Liquid Scintillation Counter	High Throughput Sensitive	Radioactive
BrdU Assay	Incorporate in Dividing Cells	Flow Cytometry Colorimetric Method	Specific and Direct High Specificity	Radioactive
⁵¹Cr Assay	Chromium-51 Release	Gamma Counter Liquid Scintillation Counter	Sensitive Reflect Cytotoxicity	Radioactivity

T Cell Suppression Assay

Regulatory T cells (Treg) are important immune cells that can control immune activation, maintain the balance of the host immune system, and prevent self-immune responses. It has been demonstrated that antigen-induced Treg cells with higher CD25 expression are derived from the same clone with corresponding CD25^low effector T cells (Teff) and possess the same epitope-specific TCR sequences.

At Creative Biolabs, the epitope-specific Treg and Teff cells can be analyzed and sorted from epitope-peptide stimulated T cells using the flow cytometric technique. For the assessment of T cell suppression, cytokine release assay (IL-2, IL-10, and IFNg) and ³H-thymidine- and CSFE-based cell proliferation assays are available.

Contact Us

Our experts are pleased to apply our professional knowledge to your program and meet your special requirements. Want to explore cancer epitopes using these versatile in vitro assays? Please contact us by email.

Reference

Kim, H. J.; Cantor, H. CD4 T-cell subsets and tumor immunity: the helpful and the not-so-helpful. Cancer immunology research. 2014, 2(2): 91-98.

For Research Use Only | Not For Clinical Use