In vitro and in vivo antitumor activity of a mouse CTL hybridoma expressing chimeric receptors bearing the single chain Fv from HER-2/neu- specific antibody and the gamma-chain from Fc(epsilon) RI.
To broaden the applicability of adoptive cellular immunotherapy against HER-2/neu overexpressing human cancers, we constructed a chimeric scFv/gamma gene composed of the variable regions of a HER-2/neu specific monoclonal antibody (mAb) joined to the signaling gamma-chain of the Fc(epsilon)RI receptor. The scFv(anti-HER-2/neu)/gamma chimeric gene was successfully expressed as functional surface receptor in the MD.45 cytolytic T-cell (CTL) hybridoma (MD.45-HER/gamma). Expression of the chimeric protein triggered IL-2 and IFN-gamma secretion in vitro upon encountering cell surface HER-2/neu and mediated non-major-histocompatibility-complex (MHC)-restricted HER-2/neu-specific target cell lysis. We also examined the in vivo activity of the MD.45-HER/gamma transduced cells. Severe combined immunodeficiency disease (SCID) mice that were given HER-2/neu positive (+) human tumor cell lines had significantly increased survival compared to mice treated with saline only, or with MD.45 cells transduced with a control anti-trinitrophenyl (anti-TNP) chimeric receptor gene (MD.45-TNP/gamma). These results demonstrate the feasibility of redirecting MD.45 CTL to react in vitro and in vivo with a variety of HER-2/neu(+) tumor cells by our gene transduction protocol. Moreover, they open the possibility of using the same chimeric gene for transducing primary lymphocytes and thus allowing adoptive immunotherapy against HER-2/neu(+) cancers.
Mamalaki
Avgi
A
Department of Biochemistry, Hellenic Pasteur Institute, Athens, Greece.
Gritzapis
Angelos D
AD
Kretsovali
Androniki
A
Belimezi
Maria
M
Papamatheakis
Joseph
J
Perez
Sonia A
SA
Papamichail
Michael
M
Baxevanis
Constantin N
CN
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D003602:Cytotoxicity, Immunologic; D006825:Hybridomas / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000235:genetics*; D007142:Immunoglobulin gamma-Chains / Q000235:genetics*; D007167:Immunotherapy; D007371:Interferon-gamma / Q000378:metabolism; D007376:Interleukin-2 / Q000378:metabolism; D008546:Melanoma, Experimental / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D051379:Mice; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D008969:Molecular Sequence Data; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology*; D017455:Receptors, IgE / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D012878:Skin Neoplasms / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D013601:T-Lymphocytes / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D007142:Immunoglobulin gamma-Chains; D007371:Interferon-gamma; D007376:Interleukin-2; D018719:Erb-b2 Receptor Tyrosine Kinases; D017455:Receptors, IgE; D011993:Recombinant Fusion Proteins; C055966:immunoglobulin Fv