Primary Polyclonal Human T Lymphocytes Targeted to Carcino-Embryonic Antigens and Neural Cell Adhesion Molecule Tumor Antigens by CD3ζ-Based Chimeric Immune Receptors

Abstract

Antigen-specific T lymphocytes are attractive as potential anticancer agents. The generation of large numbers of antigen-specific T cells is possible through the use of gene therapy to express targeting receptors on the T lymphocyte. Activated T lymphocytes were transduced to express carcino-embryonic antigen or neural cell adhesion molecule targeted CD3zeta chimeric immune receptors. The chimeric receptors were expressed as homodimers and also as heterodimers with the native CD3zeta. T lymphocyte populations were expanded in the absence of selection for the modified cells and were shown to produce cytokines when cultured in the presence of immobilized purified protein antigen. These lymphocytes also responded by cytokine production and cytolytic activity when challenged with tumor-cell lines expressing the antigen recognized by the chimeric immune receptor. The cytolytic activity appears to be largely perforin mediated. Furthermore, soluble carcino-embryonic antigen did not interfere with the functional activity of the carcino-embryonic antigen-targeted lymphocytes. Long-term (5-day) stimulation of the modified lymphocytes by protein antigen resulted in reduced viability similar to that induced by anti-CD3 antibodies alone. Viability was improved by a costimulatory signal indicating that such signals may be vital in the maintenance of long-term functional activity of receptor modified T lymphocytes.

Expression of the 5T4 oncofoetal antigen in renal cell carcinoma: a potential target for T-cell-based immunotherapy

“…The retroviral vector rKat.MFE.CD3z.TDGA.IRES.eGFP (MFE.CD3z) has been previously described (Gilham et al, 2002) and encodes a chimeric Tcell receptor specific for CEA. To create a chimeric T-cell receptor specific for the 5T4 protein, DNA coding for a humanised scFv derived from the mouse anti-human 5T4 monoclonal antibody, together with a human Fc antibody spacer region was exchanged into the above vector.…”

Tumor-specific dendritic cells generated by genetic redirection of Toll-like receptor signaling against the tumor-associated antigen, erbB2

“…ErbB2 is a cell surface growth factor receptor overexpressed on 30% of breast and ovarian cancers, but other single-chain antibody specificities exist with which it should be possible to target almost 100% of malignancies. [24][25][26][27][28][29][30] The immunotherapeutic application of this approach would involve the introduction of tumor-specific DCs into tumor-bearing hosts either alone to stimulate endogenous immune elements, or in combination with vaccines or adoptively transferred T cells that provide an enhanced number of antigen-specific T cells.…”

Ligation of the CD2 co-stimulatory receptor enhances IL-2 production from first-generation chimeric antigen receptor T cells

“…8,26 Lentivirus was produced by transient transfection of 293T cells in 150 mm 3 dishes by calcium chloride precipitation. 27 Vectors were added in the ratio 20 mg pSicoR, 13 mg pCMVDR8.91, 7 mg pMD2.g. Lentiviral particles were purified by ultracentrifugation at 71935 g, 4 1C, 2 h (Sorvall Ultra pro 80).…”