title: T lymphocytes engineered to express a CD16-chimeric antigen receptor redirect T-cell immune responses against immunoglobulin G-opsonized target cells.
abstract: OBJECTIVE
  Chimeric antigen receptors (CARs) designed for adoptive immunotherapy need to achieve two functions: antigen recognition and triggering of the lytic machinery of reprogrammed effector cells. Cytotoxic T cells have been engineered with FcγRIII (CD16) chimeric molecules to be redirected against malignant cells by monoclonal antibodies (mAbs). These cells have been proven to mediate granule-dependent cellular cytotoxicity, but it is not clear whether they can also kill malignant cells by a granule-independent mechanism of cell cytotoxicity.
  
  
  METHODS
  We engineered a CD16A-CAR equipped with the extracellular CD16A, the hinge spacer and the transmembrane region of CD8, and the ζ-chain of the T-cell receptor/CD3 complex in tandem with the CD28 co-stimulatory signal transducer module. The CD16A-CAR was expressed and functionally tested in the MD45 cell line, a murine T-cell hybridoma with a defective granular exocytosis pathway but capable of killing target cells by a Fas ligand-mediated lysis.
  
  
  RESULTS
  Our results indicate that in vitro cross-linking of CD16A-CAR on MD45 cells by the Fc fragment of mAb opsonized tumor cells induced interleukin-2 release and granule-independent cellular cytotoxicity.
  
  
  CONCLUSIONS
  We conclude that strategies aimed to implement the therapeutic functions of mAbs used in the clinic with T-dependent immune responses driven by engineered T cells expressing FcγR-CAR can boost the antitumor efficacy of mAbs used in the clinic.
authors.affiliation: Department of Clinical and Molecular Medicine, Sapienza University of Rome, Rome, Italy.
authors.forename: Maria Michela
authors.initials: MM
authors.lastname: D'Aloia
authors.affiliation: Department of Biomedicine, Institute of Translational Pharmacology, CNR, Rome, Italy; Laboratory of Molecular Medicine, Department of Systems Medicine, Tor Vergata University, Rome, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Caratelli
authors.affiliation: Department of Clinical Sciences and Translational Medicine, Tor Vergata University, Rome, Italy.
authors.forename: Camilla
authors.initials: C
authors.lastname: Palumbo
authors.affiliation: Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy.
authors.forename: Simone
authors.initials: S
authors.lastname: Battella
authors.affiliation: Laboratory of Molecular Medicine, Department of Systems Medicine, Tor Vergata University, Rome, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Arriga
authors.affiliation: Laboratory of Molecular Medicine, Department of Systems Medicine, Tor Vergata University, Rome, Italy.
authors.forename: Davide
authors.initials: D
authors.lastname: Lauro
authors.affiliation: Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Palmieri
authors.affiliation: Department of Biomedicine, Institute of Translational Pharmacology, CNR, Rome, Italy. Electronic address: giuseppe.sconocchia@cnr.it.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Sconocchia
authors.affiliation: Department of Clinical and Molecular Medicine, Sapienza University of Rome, Rome, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Alimandi
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D016827:CD8 Antigens / Q000235:genetics / Q000276:immunology; D002460:Cell Line; D003602:Cytotoxicity, Immunologic / Q000276:immunology*; D053222:Fas Ligand Protein / Q000378:metabolism; D058851:GPI-Linked Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D016219:Immunotherapy, Adoptive / Q000379:methods; D007376:Interleukin-2 / Q000378:metabolism*; D051379:Mice; D011946:Receptors, Antigen / Q000378:metabolism; D017452:Receptors, IgG / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D016827:CD8 Antigens; C505661:FASLG protein, human; C496124:FCGR3A protein, human; C428200:FCGR3B protein, human; D053222:Fas Ligand Protein; D058851:GPI-Linked Proteins; D007074:Immunoglobulin G; D007376:Interleukin-2; D011946:Receptors, Antigen; D017452:Receptors, IgG
keywords: CD16; CD28; Fas ligand; FcγR; MD45 cells; TCR; chimeric antigen receptors; ζ-chain