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Medical and Neuro-Oncology

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Le résumé fourni par la source

BACKGROUND: We demonstrated that GBM patients' T cells expressing HER2-specific chimeric antigen receptors (CAR) killed autologous HER2 + GBM cells and induced regression of autologous xenografts; however, tumors recurred in a number of treated animals, likely due to limited T-cell persistence and/or tumor antigen loss variants.Since the majority of GBMs express CMVpp65, we expressed HER2.CARs in CMV-specific cytotoxic T cells (CTLs) to enhance their persistence following native T-cell receptor (alphabetaTCR) engagement with CMV-antigens and improve their antitumor activity by simultaneously targeting CMV and HER2.METHODS: To study the feasibility of generating CMV CTLs from GBM patients, we determined the precursor frequency of CMV-specific T cells in 25 patients using IFN-gamma-ELIspot assays.To generate CMV CTL, blood mononuclear cells were transduced with an adenovirus vector, encoding the immunodominant CMV-pp65 antigen (Ad5f35pp65) and then stimulated with autologous EBV-transformed Ad5f35pp65-transduced irradiated lymphoblastic cell lines (EBV-LCL).CTLs were then transduced with a retroviral vector encoding a HER2.CAR.We detected the expression of the CAR using flow cytometry.To concomitantly identify the CMV-specific T cell population, we used pentamers for CMVpp65 and performed 51 Cr-release assays to test their functionality.RESULTS: CMV-specific T cells were readily detectable in the blood of GBM patients, as judged by IFN-gamma secretion after stimulation with CMVpp65-PepMixes TM .We were able to generate CTLs that are specific for CMV through their endogenous alphabetaTCR and specific for HER2 through CAR.Approximately 50% of CMVpp65.CTLs expressed HER2.CARs on the cell surface.CMV.HER2.CAR CTLs recognized and killed the HER2+GBM cells as well as autologous T cells pulsed with CMVpp65-PepMixes TM in 51 Cr-release assays.Neither the HER2 cell lines nor autologous T cells were killed.CONCLUSION: Our data demonstrate the feasibility of generating CAR-expressing CMVpp65specific CTLs that are functional against HER2 and CMV expressed in GBM.

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Le contrôle bibliographique ouvert

DOI retrouvé dans Crossref DOI retrouvé ; titre concordant.

Titre Crossref
Medical and Neuro-Oncology
Date Crossref
21/10/2010
Éditeur
Oxford University Press (OUP)
Type
journal-article

Ce recoupement confirme des métadonnées liées au DOI. Il ne confirme ni la méthode ni les conclusions de l’étude, et il ne compte pas comme une seconde source scientifique indépendante.

Les institutions déclarées

Une affiliation ne permet pas de déduire la nationalité d’un auteur.

Les sujets associés

Cancer Treatment and PharmacologyGlioma Diagnosis and TreatmentNeuroblastoma Research and Treatments

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