EBV-specific T cell receptor gene transfer to target nasopharyngeal carcinoma

Y Zheng, Lee Machado, B Johnson, C James, Greg Parsonage, S P Lee

Research output: Contribution to conference typesAbstract

Abstract

Infusing EBV-specific T cells is an effective prophylactic and therapeutic treatment for EBV post transplant lymphoproliferative disease. However, extending this approach to treat NPC necessitates targeting viral antigens that are subdominant targets for T cells. Therefore we have explored the use of T cell receptor (TCR) gene transfer to rapidly and reliably generate such T cell responses from all patients. To ensure a widely applicable treatment we cloned the genes encoding the TCR from an LMP2- specific T cell that is restricted through HLA A*1101, an allele carried by >50% of the Chinese population. Genes encoding the TCR α and β chains were cloned into a single retroviral vector, separated by a self-cleaving 2A peptide to ensure equal expression. T cells were transduced with this retrovirus and within 3-5 days HLA:peptide pentamer staining detected the transferred TCR in 12-17% of CD8+ T cells and 7-12% of CD4+ T cells. TCR-transduced T cells expanded rapidly in vitro in response to antigen and showed high avidity for the target peptide (10-10M) in assays of interferon-γ release. To improve safety and efficacy, the TCR was modified by codon optimization and introduction of an additional disulphide-bond. This increased almost two-fold the proportion of T cells expressing the receptor. TCR-transduced CD4+ T cells produced multiple cytokines (including IL2) in response to LMP2 suggesting they could provide a helper function in vivo to aid persistence of CD8+ effectors. Both CD8+ and CD4+ transduced T cells lysed an A11+ NPC cell line expressing LMP2.
Original languageEnglish
Publication statusPublished - 1 Jan 2010
Event14th Biennial Conference of the International Association for Research on Epstein-Barr Virus and Associated Diseases - University of Birmingham
Duration: 1 Jan 2010 → …

Conference

Conference14th Biennial Conference of the International Association for Research on Epstein-Barr Virus and Associated Diseases
Period1/01/10 → …

Fingerprint

T-Cell Receptor Genes
Human Herpesvirus 4
T-Lymphocytes
T-Cell Antigen Receptor
Peptides
Nasopharyngeal carcinoma
HLA-A Antigens
Viral Antigens
Retroviridae
Codon
Disulfides
Interferons
Interleukin-2
Therapeutics
Alleles
Staining and Labeling
Cytokines
Transplants
Safety

Cite this

Zheng, Y., Machado, L., Johnson, B., James, C., Parsonage, G., & Lee, S. P. (2010). EBV-specific T cell receptor gene transfer to target nasopharyngeal carcinoma. Abstract from 14th Biennial Conference of the International Association for Research on Epstein-Barr Virus and Associated Diseases, .
Zheng, Y ; Machado, Lee ; Johnson, B ; James, C ; Parsonage, Greg ; Lee, S P. / EBV-specific T cell receptor gene transfer to target nasopharyngeal carcinoma. Abstract from 14th Biennial Conference of the International Association for Research on Epstein-Barr Virus and Associated Diseases, .
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abstract = "Infusing EBV-specific T cells is an effective prophylactic and therapeutic treatment for EBV post transplant lymphoproliferative disease. However, extending this approach to treat NPC necessitates targeting viral antigens that are subdominant targets for T cells. Therefore we have explored the use of T cell receptor (TCR) gene transfer to rapidly and reliably generate such T cell responses from all patients. To ensure a widely applicable treatment we cloned the genes encoding the TCR from an LMP2- specific T cell that is restricted through HLA A*1101, an allele carried by >50{\%} of the Chinese population. Genes encoding the TCR α and β chains were cloned into a single retroviral vector, separated by a self-cleaving 2A peptide to ensure equal expression. T cells were transduced with this retrovirus and within 3-5 days HLA:peptide pentamer staining detected the transferred TCR in 12-17{\%} of CD8+ T cells and 7-12{\%} of CD4+ T cells. TCR-transduced T cells expanded rapidly in vitro in response to antigen and showed high avidity for the target peptide (10-10M) in assays of interferon-γ release. To improve safety and efficacy, the TCR was modified by codon optimization and introduction of an additional disulphide-bond. This increased almost two-fold the proportion of T cells expressing the receptor. TCR-transduced CD4+ T cells produced multiple cytokines (including IL2) in response to LMP2 suggesting they could provide a helper function in vivo to aid persistence of CD8+ effectors. Both CD8+ and CD4+ transduced T cells lysed an A11+ NPC cell line expressing LMP2.",
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Zheng, Y, Machado, L, Johnson, B, James, C, Parsonage, G & Lee, SP 2010, 'EBV-specific T cell receptor gene transfer to target nasopharyngeal carcinoma' 14th Biennial Conference of the International Association for Research on Epstein-Barr Virus and Associated Diseases, 1/01/10, .

EBV-specific T cell receptor gene transfer to target nasopharyngeal carcinoma. / Zheng, Y; Machado, Lee; Johnson, B; James, C; Parsonage, Greg; Lee, S P.

2010. Abstract from 14th Biennial Conference of the International Association for Research on Epstein-Barr Virus and Associated Diseases, .

Research output: Contribution to conference typesAbstract

TY - CONF

T1 - EBV-specific T cell receptor gene transfer to target nasopharyngeal carcinoma

AU - Zheng, Y

AU - Machado, Lee

AU - Johnson, B

AU - James, C

AU - Parsonage, Greg

AU - Lee, S P

PY - 2010/1/1

Y1 - 2010/1/1

N2 - Infusing EBV-specific T cells is an effective prophylactic and therapeutic treatment for EBV post transplant lymphoproliferative disease. However, extending this approach to treat NPC necessitates targeting viral antigens that are subdominant targets for T cells. Therefore we have explored the use of T cell receptor (TCR) gene transfer to rapidly and reliably generate such T cell responses from all patients. To ensure a widely applicable treatment we cloned the genes encoding the TCR from an LMP2- specific T cell that is restricted through HLA A*1101, an allele carried by >50% of the Chinese population. Genes encoding the TCR α and β chains were cloned into a single retroviral vector, separated by a self-cleaving 2A peptide to ensure equal expression. T cells were transduced with this retrovirus and within 3-5 days HLA:peptide pentamer staining detected the transferred TCR in 12-17% of CD8+ T cells and 7-12% of CD4+ T cells. TCR-transduced T cells expanded rapidly in vitro in response to antigen and showed high avidity for the target peptide (10-10M) in assays of interferon-γ release. To improve safety and efficacy, the TCR was modified by codon optimization and introduction of an additional disulphide-bond. This increased almost two-fold the proportion of T cells expressing the receptor. TCR-transduced CD4+ T cells produced multiple cytokines (including IL2) in response to LMP2 suggesting they could provide a helper function in vivo to aid persistence of CD8+ effectors. Both CD8+ and CD4+ transduced T cells lysed an A11+ NPC cell line expressing LMP2.

AB - Infusing EBV-specific T cells is an effective prophylactic and therapeutic treatment for EBV post transplant lymphoproliferative disease. However, extending this approach to treat NPC necessitates targeting viral antigens that are subdominant targets for T cells. Therefore we have explored the use of T cell receptor (TCR) gene transfer to rapidly and reliably generate such T cell responses from all patients. To ensure a widely applicable treatment we cloned the genes encoding the TCR from an LMP2- specific T cell that is restricted through HLA A*1101, an allele carried by >50% of the Chinese population. Genes encoding the TCR α and β chains were cloned into a single retroviral vector, separated by a self-cleaving 2A peptide to ensure equal expression. T cells were transduced with this retrovirus and within 3-5 days HLA:peptide pentamer staining detected the transferred TCR in 12-17% of CD8+ T cells and 7-12% of CD4+ T cells. TCR-transduced T cells expanded rapidly in vitro in response to antigen and showed high avidity for the target peptide (10-10M) in assays of interferon-γ release. To improve safety and efficacy, the TCR was modified by codon optimization and introduction of an additional disulphide-bond. This increased almost two-fold the proportion of T cells expressing the receptor. TCR-transduced CD4+ T cells produced multiple cytokines (including IL2) in response to LMP2 suggesting they could provide a helper function in vivo to aid persistence of CD8+ effectors. Both CD8+ and CD4+ transduced T cells lysed an A11+ NPC cell line expressing LMP2.

M3 - Abstract

ER -

Zheng Y, Machado L, Johnson B, James C, Parsonage G, Lee SP. EBV-specific T cell receptor gene transfer to target nasopharyngeal carcinoma. 2010. Abstract from 14th Biennial Conference of the International Association for Research on Epstein-Barr Virus and Associated Diseases, .