一种用于转移性黑色素瘤过继性免疫治疗的选择和扩增表位特异性T淋巴细胞的完整GMP流程。
A full GMP process to select and amplify epitope-specific T lymphocytes for adoptive immunotherapy of metastatic melanoma.
作者信息
Labarriere N, Fortun A, Bellec A, Khammari A, Dreno B, Saïagh S, Lang F
机构信息
Inserm, U892, 44000 Nantes, France ; Université de Nantes, 44000 Nantes, France ; CNRS, UMR 6299, 44000 Nantes, France.
出版信息
Clin Dev Immunol. 2013;2013:932318. doi: 10.1155/2013/932318. Epub 2013 Sep 30.
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.
在过去20年里,已经对转移性黑色素瘤患者进行了多项肿瘤特异性T淋巴细胞过继转移试验,随着相关黑色素瘤抗原的确定以及注入T细胞的纯度/特异性提高,试验结果越来越令人鼓舞。我们之前描述了一种使用包被有HLA/肽复合物的磁珠来分选表位特异性T淋巴细胞的方法,并认为该方法可应用于临床。在本研究中,我们详细描述了用于分选和扩增针对黑色素瘤抗原Melan - A和MELOE - 1的临床级T细胞的整个GMP过程。该过程中使用的所有试剂(包括分选试剂)均在GMP条件下生产,并且我们记录了该过程不同步骤(如肽刺激、分选和扩增)的优化情况。在临床环境中进行的3次空白试验验证了该优化程序,从转移性黑色素瘤患者的100 mL血液开始,可在28天内产生至少10⁸个纯的(>90%)Melan - A和MELOE - 1特异性T细胞。因此,这一GMP过程已准备好在即将开展的针对转移性黑色素瘤患者的I/II期临床试验中使用。
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