T cells expressing VHH-directed oligoclonal chimeric HER2 antigen receptors: Towards tumor-directed oligoclonal T cell therapy

• Published in: Biochimica et biophysica acta Vol. 1840; no. 1; pp. 378 - 386
• Main Authors: Jamnani, Fatemeh Rahimi, Rahbarizadeh, Fatemeh, Shokrgozar, Mohammad Ali, Mahboudi, Fereidoun, Ahmadvand, Davoud, Sharifzadeh, Zahra, Parhamifar, Ladan, Moghimi, S. Moein
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.01.2014
• Subjects: antibodies; antigens; Apoptosis; binding capacity; Blotting, Western; Camelidae; camels; Cell Proliferation; Cell- and Tissue-Based Therapy; Chimeric antigen receptor; chimerism; clones; cytokines; cytotoxicity; Flow Cytometry; genetic engineering; HER2; Humans; immunotherapy; Jurkat Cells; neoplasm cells; neoplasms; Neoplasms - immunology; Neoplasms - metabolism; Neoplasms - therapy; Oligoclonal T cell therapy; Protein Engineering; Receptor, ErbB-2 - genetics; Receptor, ErbB-2 - immunology; Receptor, ErbB-2 - metabolism; receptors; Receptors, Antigen - genetics; Receptors, Antigen - immunology; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - immunology; Recombinant Fusion Proteins - metabolism; secretion; Single domain antibodies (VHH); Single-Domain Antibodies - immunology; T-lymphocytes; T-Lymphocytes - immunology; T-Lymphocytes - metabolism; Tumor Cells, Cultured; TCR; TMB; MHC; TAG 72; Oligoclonal T cell therapy; Single domain antibodies (VHH); CDR; Chimeric antigen receptor; CAR; VHH; IPTG; HER2; HCAb; VHH-CAR; T cell receptor; variable domain of camel heavy-chain antibody; heavy-chain antibodies; tumor associated glycoprotein 72; isopropyl-β-D-thio-galactoside; major histocompatibility complex; 3,3′,5,5′-tetramethyl benzidine; VHH-chimeric antigen receptor; complementarity determining region
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2013.09.029

Adoptive cell therapy with engineered T cells expressing chimeric antigen receptors (CARs) originated from antibodies is a promising strategy in cancer immunotherapy. Several unsuccessful trials, however, highlight the need for alternative conventional binding domains and the better combination of costimulatory endodomains for CAR construction to improve the effector functions of the engineered T cells. Camelid single-domain antibodies (VHHs), which are the smallest single domain antibodies, can endow great targeting ability to CAR-engineered T cells. We have developed a method to generate genetically engineered Jurkat T cells armed with a CAR comprising the anti-HER2 VHH as targeting moiety. From an immune camel library, five VHH clones were selected as a set of oligoclonal anti-HER2 VHHs that exhibited diverse binding abilities and joined them to CD28-CD3ζ and CD28-OX40-CD3ζ signaling endodomains. Jurkat T cells expression of VHH-CARs and cell functions were evaluated. The oligoclonal engineered T cells showed higher proliferation, cytokine secretion and cytotoxicity than each individual VHH-CAR-engineered Jurkat T cells. The combination of superior targeting ability of oligoclonal VHHs with the third generation CAR can substantially improve the function of engineered T cells. Antigen-specific directed oligoclonal T cells are alternatively promising, but safer systems, to combat tumor cells. •Five epitope-distinct VHHs with different binding abilities to HER2 were identified.•Anti-HER2 VHHs were used as targeting moieties for generating engineered T cells.•T cells expressing VHH-directed oligoclonal chimeric HER2 antigen receptors were investigated.•The oligoclonal engineered T cells function better than each individual VHH-CAR-engineered T cells.