Dataset Information

Identification of Functional HLA-A*01 :01-Restricted EBV-LMP2-Specific T-cell Receptors.

ABSTRACT:

Background

Adoptive transfer of genetically engineered T cells expressing antigen-specific T-cell receptors (TCRs), is an appealing therapeutic approach for Epstein-Barr virus (EBV)-associated malignancies of latency type II/III that express EBV-antigens (LMP1/2). Patients who are HLA-A*01:01pos could benefit from such products, since no T cells recognizing any EBV-derived peptide in this common HLA allele have been found thus far.

Methods

HLA-A*01:01-restricted EBV-(LMP2)-specific T-cells were isolated using peptide-MHC-tetramers. Functionality was assessed by production of IFNγ and cytotoxicity when stimulated with EBV-LMP2-expressing cell-lines. Functionality of primary T cells transduced with HLA-A*01:01-restricted EBV-LMP2-specific TCRs was optimized by knocking out the endogenous TCR of primary T cells (ΔTCR) using CRISPR-Cas9 technology.

Results

EBV-LMP2-specific T cells were successfully isolated and their TCRs were characterized. TCR gene-transfer in primary T cells resulted in specific peptide-MHC-tetramer binding and reactivity against EBV-LMP2-expressing cell-lines. The mean-fluorescence intensity of peptide-MHC-tetramer binding was increased 1.5-2 fold when the endogenous TCR of CD8pos T cells was knocked out. CD8pos/ΔTCR T cells modified to express EBV-LMP2-specific TCRs showed IFNγ secretion and cytotoxicity towards EBV-LMP2-expressing malignant cell-lines.

Discussion

We isolated the first functional HLA-A*01:01-restricted EBV-LMP2-specific T-cell populations and TCRs, which can potentially be used in future TCR gene-therapy to treat EBV-associated latency type II/III malignancies.

SUBMITTER: Huisman W

PROVIDER: S-EPMC9470112 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature

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Publications

Identification of Functional HLA-A*01:01-Restricted Epstein-Barr Latent Membrane Protein 2-Specific T-Cell Receptors.

Huisman Wesley W Gille Ilse I van der Maarel Lieve E LE Hageman Lois L Morton Laura T LT de Jong Rob C M RCM Heemskerk Mirjam H M MHM Amsen Derk D Falkenburg J H Frederik JHF Jedema Inge I

The Journal of infectious diseases 20220901 5

<h4>Background</h4>Adoptive transfer of genetically engineered T cells expressing antigen-specific T-cell receptors (TCRs) is an appealing therapeutic approach for Epstein-Barr virus (EBV)-associated malignancies of latency type II/III that express EBV antigens (LMP1/2). Patients who are HLA-A*01:01 positive could benefit from such products, since no T cells recognizing any EBV-derived peptide in this common HLA allele have been found thus far.<h4>Methods</h4>HLA-A*01:01-restricted EBV-LMP2-spec ...[more]

PMID: 32808978

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