title: Peptide fine specificity of anti-glycoprotein 100 CTL is preserved following transfer of engineered TCR alpha beta genes into primary human T lymphocytes.
abstract: TCR with known antitumor reactivity can be genetically introduced into primary human T lymphocytes and provide promising tools for immunogene therapy of tumors. We molecularly characterized two distinct TCRs specific for the same HLA-A2-restricted peptide derived from the melanocyte differentiation Ag gp100, yet exhibiting different stringencies in peptide requirements. The existence of these two distinct gp100-specific TCRs allowed us to study the preservation of peptide fine specificity of native TCRalphabeta when engineered for TCR gene transfer into human T lymphocytes. Retroviral transduction of primary human T lymphocytes with either one of the two sets of TCRalphabeta constructs enabled T lymphocytes to specifically kill and produce TNF-alpha when triggered by native gp100(pos)/HLA-A2(pos) tumor target cells as well as gp100 peptide-loaded HLA-A2(pos) tumor cells. Peptide titration studies revealed that the cytolytic efficiencies of the T lymphocyte transductants were in the same range as those of the parental CTL clones. Moreover, primary human T lymphocytes expressing either one of the two engineered gp100-specific TCRs show cytolytic activities in response to a large panel of peptide mutants that are identical with those of the parental CTL. The finding that two gp100-specific TCR, derived from two different CTL, can be functionally introduced into primary human T lymphocytes without loss of the Ag reactivity and peptide fine specificity, holds great promise for the application of TCR gene transfer in cancer immunotherapy.
authors.affiliation: Clinical and Tumor Immunology, Department of Medical Oncology, Erasmus Medical Center-Daniel den Hoed, Groene Hilledijk 301, 3075 EA Rotterdam, The Netherlands.
authors.forename: Niels
authors.initials: N
authors.lastname: Schaft
authors.forename: Ralph A
authors.initials: RA
authors.lastname: Willemsen
authors.forename: Jolanda
authors.initials: J
authors.lastname: de Vries
authors.forename: Birgit
authors.initials: B
authors.lastname: Lankiewicz
authors.forename: Bram W L
authors.initials: BW
authors.lastname: Essers
authors.forename: Jan-Willem
authors.initials: JW
authors.lastname: Gratama
authors.forename: Carl G
authors.initials: CG
authors.lastname: Figdor
authors.forename: Reinier L H
authors.initials: RL
authors.lastname: Bolhuis
authors.forename: Reno
authors.initials: R
authors.lastname: Debets
authors.forename: Gosse J
authors.initials: GJ
authors.lastname: Adema
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D002460:Cell Line; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D002999:Clone Cells; D003601:Cytotoxicity Tests, Immunologic; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology*; D015789:HLA-A2 Antigen / Q000276:immunology; D006801:Humans; D020014:K562 Cells; D008545:Melanoma / Q000276:immunology / Q000628:therapy; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics / Q000276:immunology*; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000096:biosynthesis / Q000235:genetics* / Q000627:therapeutic use; D016176:T-Lymphocyte Subsets / Q000276:immunology* / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods*; D014162:Transfection; D014407:Tumor Cells, Cultured; D058951:gp100 Melanoma Antigen
chemical_list: D018984:Epitopes, T-Lymphocyte; D015789:HLA-A2 Antigen; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; C548637:PMEL protein, human; D010446:Peptide Fragments; D016693:Receptors, Antigen, T-Cell, alpha-beta; D058951:gp100 Melanoma Antigen