title: In Vitro and In Vivo Comparison of Lymphocytes Transduced with a Human CD16 or with a Chimeric Antigen Receptor Reveals Potential Off-Target Interactions due to the IgG2 CH2-CH3 CAR-Spacer.
abstract: The present work was designed to compare two mechanisms of cellular recognition based on Ab specificity: firstly, when the anti-HER2 mAb trastuzumab bridges target cells and cytotoxic lymphocytes armed with a Fc receptor (ADCC) and, secondly, when HER2 positive target cells are directly recognized by cytotoxic lymphocytes armed with a chimeric antigen receptor (CAR). To compare these two mechanisms, we used the same cellular effector (NK-92) and the same signaling domain (FcεRIγ). The NK-92 cytotoxic cell line was transfected with either a FcγRIIIa-FcεRIγ (NK-92(CD16)) or a trastuzumab-based scFv-FcεRIγ chimeric receptor (NK-92(CAR)). In vitro, the cytotoxic activity against HER2 positive target cells after indirect recognition by NK-92(CD16) was always inferior to that observed after direct recognition by NK-92(CAR). In contrast, and somehow unexpectedly, in vivo, adoptive transfer of NK-92(CD16) + trastuzumab but not of NK-92(CAR) induced tumor regression. Analysis of the in vivo xenogeneic system suggested that the human CH2-CH3 IgG2 used as a spacer in our construct was able to interact with the FcR present at the cell surface of the few NSG-FcR+ remaining immune cells. This interaction, leading to blockage of the NK-92(CAR) in the periphery of the engrafted tumor cells, stresses the critical role of the composition of the spacer domain.
authors.affiliation: UMR INSERM U892, 8 Quai Moncousu, 44007 Nantes Cedex, France ; Centre Hospitalier Universitaire de Nantes, 1 Place Ricordeau, 44000 Nantes, France.
authors.forename: Béatrice
authors.initials: B
authors.lastname: Clémenceau
authors.affiliation: UMR INSERM 1052, Centre Léon Bérard, 28 rue Laennec, 69008 Lyon, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Valsesia-Wittmann
authors.affiliation: UMR INSERM 1052, Centre Léon Bérard, 28 rue Laennec, 69008 Lyon, France.
authors.forename: Anne-Catherine
authors.initials: AC
authors.lastname: Jallas
authors.affiliation: UMR INSERM U892, 8 Quai Moncousu, 44007 Nantes Cedex, France.
authors.forename: Régine
authors.initials: R
authors.lastname: Vivien
authors.affiliation: Genenthec Inc., South San Francisco, CA 94080, USA.
authors.forename: Raphaël
authors.initials: R
authors.lastname: Rousseau
authors.affiliation: UMR INSERM 1052, Centre Léon Bérard, 28 rue Laennec, 69008 Lyon, France.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Marabelle
authors.affiliation: UMR INSERM 1052, Centre Léon Bérard, 28 rue Laennec, 69008 Lyon, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Caux
authors.affiliation: UMR INSERM U892, 8 Quai Moncousu, 44007 Nantes Cedex, France.
authors.forename: Henri
authors.initials: H
authors.lastname: Vié
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D000068818:Cetuximab / Q000737:chemistry / Q000494:pharmacology; D003602:Cytotoxicity, Immunologic / Q000187:drug effects*; D058851:GPI-Linked Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D020014:K562 Cells; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D017455:Receptors, IgE / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D017452:Receptors, IgG / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology / Q000276:immunology*; D014161:Transduction, Genetic; D000068878:Trastuzumab / Q000737:chemistry / Q000494:pharmacology; D023041:Xenograft Model Antitumor Assays
chemical_list: D000970:Antineoplastic Agents; D000068818:Cetuximab; D058851:GPI-Linked Proteins; D007074:Immunoglobulin G; D018719:Erb-b2 Receptor Tyrosine Kinases; D017455:Receptors, IgE; D017452:Receptors, IgG; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; D000068878:Trastuzumab; C496124:FCGR3A protein, human; C428200:FCGR3B protein, human; C543372:FcepsilonRI gamma-chain, human; C508053:ERBB2 protein, human