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Last Updated: 27/08/2024

Validation of Plasmodium falciparum antigens targeted by human CD8+ T cells using T Cell Receptor-expressing reporter cells

Objectives

The objective of this project is to use Cellular Indexing of Transcriptomes and Epitopes by Sequencing (CITE-Seq) to single-cell sequence cytotoxic T lymphocytes (CTLs) and validate Plasmodium falciparum antigen-specific T cells (Pf ATCs) through specific T-cell receptor (TCR) activation in reporter cells.

Principal Investigators / Focal Persons

Helder Takashi Imoto Nakaya

Rationale and Abstract

In collaboration with Dr. Helder Nakaya from the Scientific Platform Pasteur USP, Brazil, an expert in systems vaccinology and single-cell immunoprofiling, Cellular Indexing of Transcriptomes and Epitopes by Sequencing (CITE-Seq) will be used to single-cell sequence cytotoxic T lymphocytes (CTLs) using barcoded dextramers harboring the epitopes discovered by immunopeptidomics. This method will allow the small numbers of specific T cells from the humanized mice to be maximized in order to retrieve the sequences of epitope-specific variable regions of T cell receptors (TCRs). The TCRs will be expressed on reporter cells, which will become fluorescent after TCR activation by the complex peptide-MHC class I molecule. Therefore, instead of focusing on the elimination of infected hepatocytes, which can depend on non-specific factors, such as secreted cytokines in vitro or on the high amounts of CTLs that will be determined by the immunization method/model in vivo, validation of Pf ATCs will be proposed by relying on the specific TCR activation of reporter cells by epitopes presented on the surface of Pf infected hepatocytes.

Thematic Categories

Basic Science

Date

Jan 2023

Project Site

Brazil

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