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Clinical and Developmental Immunology
Volume 2013 (2013), Article ID 932318, 11 pages
http://dx.doi.org/10.1155/2013/932318
Research Article

A Full GMP Process to Select and Amplify Epitope-Specific T Lymphocytes for Adoptive Immunotherapy of Metastatic Melanoma

N. Labarriere,1,2,3 A. Fortun,1,2,3 A. Bellec,4 A. Khammari,1,2,3,5 B. Dreno,1,2,3,4,5 S. Saïagh,4 and F. Lang1,2,3

1Inserm, U892, 44000 Nantes, France
2Université de Nantes, 44000 Nantes, France
3CNRS, UMR 6299, 44000 Nantes, France
4Unit of Cell and Gene Therapy, CHU Nantes, 44000 Nantes, France
5Unit of Skin Cancer, CHU Nantes, 44000 Nantes, France

Received 18 July 2013; Revised 27 August 2013; Accepted 27 August 2013

Academic Editor: John Maher

Copyright © 2013 N. Labarriere et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

A number of trials of adoptive transfer of tumor-specific T lymphocytes have been performed in the last 20 years in metastatic melanoma, with increasingly encouraging results as the relevant melanoma antigens were identified and the purity/specificity of injected T cells improved. We have previously described a sorting method of epitope-specific T lymphocytes that uses magnetic beads coated with HLA/peptide complexes and we suggested that this method could be applied to a clinical setting. In the present work, we provide a detailed description of the whole GMP process of sorting and amplification of clinical grade T cells specific for the melanoma antigens Melan-A and MELOE-1. All the reagents used in this process including the sorting reagent were produced in GMP conditions and we document the optimization of the different steps of the process such as peptide stimulation, sorting, and amplification. The optimized procedure, validated in 3 blank runs in a clinical setting, allowed the production of at least 108 pure (>90%) Melan-A- and MELOE-1-specific T cells within 28 days starting with 100 mL of blood from metastatic melanoma patients. This GMP process is thus ready to be used in an upcoming phase I/II clinical trial on metastatic melanoma patients.