Influenza virus-specific TCR-transduced T cells as a model for adoptive immunotherapy

• Published in: Human vaccines & immunotherapeutics 2013-06, Vol.9 (6), p.1205-1216
• Main Authors: Berdien, Belinda, Reinhard, Henrike, Meyer, Sabrina, Spöck, Stefanie, Kröger, Nicolaus, Atanackovic, Djordje, Fehse, Boris
• Format: Article
• Language: English
• Subjects: adoptive immunotherapy; CD4-Positive T-Lymphocytes - immunology; CD8-Positive T-Lymphocytes - immunology; Gene Expression; gene transfer; Genetic Vectors; Healthy Volunteers; Humans; Immunotherapy, Adoptive - methods; influenza antigen; Interferon-gamma - secretion; Interleukin-2 - secretion; lentiviral vectors; Lentivirus - genetics; Receptors, Antigen, T-Cell - genetics; Receptors, Antigen, T-Cell - immunology; Research Paper; RNA-Binding Proteins - immunology; T-cell receptor (TCR); TCR gene therapy; Transduction, Genetic; Tumor Necrosis Factor-alpha - secretion; Viral Core Proteins - immunology; Viral Matrix Proteins - immunology
• ISSN: 2164-5515; 2164-554X
• DOI: 10.4161/hv.24051

Adoptive transfer of T lymphocytes equipped with tumor-antigen specific T-cell receptors (TCRs) represents a promising strategy in cancer immunotherapy, but the approach remains technically demanding. Using influenza virus (Flu)-specific T-cell responses as a model system we compared different methods for the generation of T-cell clones and isolation of antigen-specific TCRs. Altogether, we generated 12 CD8 + T-cell clones reacting to the Flu matrix protein (Flu-M) and 6 CD4 + T-cell clones reacting to the Flu nucleoprotein (Flu-NP) from 4 healthy donors. IFN-γ-secretion-based enrichment of antigen-specific cells, optionally combined with tetramer staining, was the most efficient way for generating T-cell clones. In contrast, the commonly used limiting dilution approach was least efficient. TCR genes were isolated from T-cell clones and cloned into both a previously used gammaretroviral LTR-vector, MP91 and the novel lentiviral self-inactivating vector LeGO-MP that contains MP91-derived promotor and regulatory elements. To directly compare their functional efficiencies, we in parallel transduced T-cell lines and primary T cells with the two vectors encoding identical TCRs. Transduction efficiencies were approximately twice higher with the gammaretroviral vector. Secretion of high amounts of IFN-γ, IL-2 and TNF-α by transduced cells after exposure to the respective influenza target epitope proved efficient specificity transfer of the isolated TCRs to primary T-cells for both vectors, at the same time indicating superior functionality of MP91-transduced cells. In conclusion, we have developed optimized strategies to obtain and transfer antigen-specific TCRs as well as designed a novel lentiviral vector for TCR-gene transfer. Our data may help to improve adoptive T-cell therapies.