Matthew Wilson
Last active: 3/24/2020

PiggyBac-mediated cancer immunotherapy using EBV-specific cytotoxic T-cells expressing HER2-specific chimeric antigen receptor.

Nakazawa Y, Huye LE, Salsman VS, Leen AM, Ahmed N, Rollins L, Dotti G, Gottschalk SM, Wilson MH, Rooney CM
Mol Ther. 2011 19 (12): 2133-43

PMID: 21772253 · PMCID: PMC3242651 · DOI:10.1038/mt.2011.131

Epstein-Barr virus (EBV)-specific cytotoxic T lymphocytes (CTLs) can be modified to function as heterologous tumor directed effector cells that survive longer in vivo than tumor directed T cells without virus specificity, due to chronic stimulation by viral antigens expressed during persistent infection in seropositive individuals. We evaluated the nonviral piggyBac (PB) transposon system as a platform for modifying EBV-CTLs to express a functional human epidermal growth factor receptor 2-specific chimeric antigen receptor (HER2-CAR) thereby directing virus-specific, gene modified CTLs towards HER2-positive cancer cells. Peripheral blood mononuclear cells (PBMCs) were nucleofected with transposons encoding a HER2-CAR and a truncated CD19 molecule for selection followed by specific activation and expansion of EBV-CTLs. HER2-CAR was expressed in ~40% of T cells after CD19 selection with retention of immunophenotype, polyclonality, and function. HER2-CAR-modified EBV-CTLs (HER2-CTLs) killed HER2-positive brain tumor cell lines in vitro, exhibited transient and reversible increases in HER2-CAR expression following antigen-specific stimulation, and stably expressed HER2-CAR beyond 120 days. Adoptive transfer of PB-modified HER2-CTLs resulted in tumor regression in a murine xenograft model. Our results demonstrate that PB can be used to redirect virus-specific CTLs to tumor targets, which should prolong tumor-specific T cell survival in vivo producing more efficacious immunotherapy.

MeSH Terms (22)

Animals Antigen Presentation Antigens, CD19 Brain Neoplasms Cell Line, Tumor Coculture Techniques Enzyme-Linked Immunosorbent Assay Epstein-Barr Virus Infections Flow Cytometry Herpesvirus 4, Human Humans Immunotherapy Male Mice Mice, Inbred NOD Mice, SCID Receptor, ErbB-2 Receptors, Antigen Recombinant Fusion Proteins T-Lymphocytes, Cytotoxic Transduction, Genetic Tumor Cells, Cultured

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