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New Technique Could Help Scientists Identify Rare T Cells and Advance T Cell Therapies

May 11, 2021 10:10:00 AM / by Stacy Matthews Branch, DVM, PhD

AdobeStock_328733368 (1)-1Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research scientists developed a new technique that enables researchers to identify and isolate T cells capable of targeting cancer cells, viruses, and other rare diseases more efficiently.

There are a number of challenges with adoptive T cell therapies, one being the isolation of specific T cells in sufficient numbers. Although state-of-the-art technologies are available to isolate rare, specific T cells, those methodologies are quite labor-intensive and tend to also lead to the isolation of bystander T cells that are activated with antigen recognition by T cells but do not themselves react to antigens. The development of a technology that bolsters the numbers of antigen-specific T cells, such a those reactive to tumor cells or viruses, is an ongoing research endeavor.

Researchers from Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research at UCLA have developed a new technique that is less labor-intensive than current methods while providing more specificity for rare antigen-specific T cells. The method involves a high-throughput mRNA sequencing technology performed at the single-cell level in cells. This is achieved via cross-linker regulated intracellular phenotype (CLInt-seq) that utilizes a reversible cross-linking technique to fix the intracellular proteins in a manner that does not degrade RNA. The technique allows the isolation of any rare T cell receptor, and the receptor identification can be based on a particular cytokine(s) or transcription factor(s).

In the study, the researchers performed intracellular staining for TNF-alpha and IFN-gamma and identified cytomegalovirus and Epstein-Barr virus reactive T cell receptors as efficiently as using the more labor-intensive peptide-MHC multimer technology. CLInt-seq also allowed the profiling of regulatory T cells based on intracellular staining of transcription factor FOXP3, a regulatory T-cell gene expression regulator. The study data further showed that intracellular staining identifies antigen-specific T cells with a lower false positive rate compared to using cell surface protein profiling.

Given the limitations overcome with CLInt-seq, the new technique can increase the knowledge base regarding T cell responses to many different types of antigens. This, in turn, can facilitate the development of novel immunotherapies. Using CLInt-seq, the UCLA research group focuses on developing T cell receptor-based therapies for prostate cancer.

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Scientists develop method to more efficiently isolate and identify rare T cells. (2021). Retrieved 12 February 2021, from

Topics: Cancer, T Cells

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