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Protocol for the detection of defined T cell clones in a heterogeneous cell population.

ABSTRACT: Identifying defined T cell clones within a polyclonal population is key to clarifying their phenotype and function. Here, we present a protocol for detecting specified T cell clones in a heterogeneous cell population. We describe steps for stimulating human CD4⁺ T cells isolated from blood with a protein antigen, sorting antigen-specific cells by fluorescence-activated cell sorting, and detecting among these the presence of predefined T cell clones, based on their T cell receptor (TCR). TCR cDNA is amplified through 5'-RACE (TCR-SMART) and detected by qPCR. For complete details on the use and execution of this protocol, please refer to Notarbartolo et al. (2021).¹.

SUBMITTER: Gobbini A

PROVIDER: S-EPMC10783555 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature

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Protocol for the detection of defined T cell clones in a heterogeneous cell population.

Gobbini Andrea A Bandera Alessandra A Grifantini Renata R Abrignani Sergio S Notarbartolo Samuele S

STAR protocols 20231221 1

Identifying defined T cell clones within a polyclonal population is key to clarifying their phenotype and function. Here, we present a protocol for detecting specified T cell clones in a heterogeneous cell population. We describe steps for stimulating human CD4<sup>+</sup> T cells isolated from blood with a protein antigen, sorting antigen-specific cells by fluorescence-activated cell sorting, and detecting among these the presence of predefined T cell clones, based on their T cell receptor (TCR ...[more]

PMID: 38141168

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