title: Langerhans cells derived from genetically modified human CD34+ hemopoietic progenitors are more potent than peptide-pulsed Langerhans cells for inducing antigen-specific CD8+ cytolytic T lymphocyte responses.
abstract: Sustained Ag expression by human dendritic cells (DCs) is an attractive means of optimizing Ag presentation for stimulating durable cellular immunity. To establish proof of principle, we used Langerhans cell (LC) progeny of retrovirally transduced CD34(+) hemopoietic progenitor cells to stimulate responses against the HLA-A*0201-restricted influenza matrix peptide (fluMP). Retroviral transduction of CD34(+) hemopoietic progenitor cells, during pre-expansion by thrombopoietin, c-kit ligand, and FLT-3 ligand, on recombinant fibronectin, but in the absence of FCS, resulted in gene expression by 20-30% of the LCs. Expression persisted at least 28 days, with little decline (<30%) over that time. Retroviral transduction did not alter the phenotype or potent immunogenicity of normal mature DCs. FluMP-transduced LCs stimulated a 130-fold expansion of T cells reactive with HLA-A*0201-fluMP tetramers, even at LC:T cell ratios of 1:100-150 and lower, whereas fluMP-pulsed LCs stimulated only a 30-fold expansion. FluMP-transduced LCs also stimulated higher IFN-gamma secretion (100-123 spot-forming cells/10(5) CD8(+) T cells) than did fluMP-pulsed LCs (10-91 spot-forming cells/10(5) CD8(+) T cells). CD8(+) T cells stimulated by transduced LCs did not react preferentially with retrovirally transduced targets, indicating that the responses targeted only the immunizing influenza and not the retroviral vector Ags, even though these could have provided nonspecific helper epitopes presented by the transduced LCs. These data demonstrate that gene-transduced LCs maintain the activated phenotype as well potent immunogenicity typical of mature DCs. LCs genetically modified to express fluMP are also more potent stimulators of Ag-specific CD8(+) T cell responses than are peptide-pulsed LCs.
authors.affiliation: Laboratory of Cellular Immunobiology, Division of Hematologic Oncology, Department of Medicine, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
authors.forename: Jianda
authors.initials: J
authors.lastname: Yuan
authors.forename: Jean-Baptiste
authors.initials: JB
authors.lastname: Latouche
authors.forename: John L
authors.initials: JL
authors.lastname: Reagan
authors.forename: Glenn
authors.initials: G
authors.lastname: Heller
authors.forename: Isabelle
authors.initials: I
authors.lastname: Riviere
authors.forename: Michel
authors.initials: M
authors.lastname: Sadelain
authors.forename: James W
authors.initials: JW
authors.lastname: Young
mesh_terms: D017951:Antigen Presentation / Q000235:genetics; D018952:Antigens, CD34 / Q000096:biosynthesis* / Q000235:genetics; D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle / Q000276:immunology; D002454:Cell Differentiation / Q000276:immunology; D003601:Cytotoxicity Tests, Immunologic; D003602:Cytotoxicity, Immunologic / Q000235:genetics*; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005353:Fibronectins / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology; D016056:Immunodominant Epitopes / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D007801:Langerhans Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*; D008213:Lymphocyte Activation / Q000235:genetics; D008565:Membrane Proteins / Q000494:pharmacology; D010455:Peptides / Q000276:immunology* / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics / Q000254:growth & development / Q000276:immunology; D019089:Stem Cell Factor / Q000494:pharmacology; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D013926:Thrombopoietin / Q000276:immunology; D014161:Transduction, Genetic / Q000379:methods
chemical_list: D018952:Antigens, CD34; D000956:Antigens, Viral; D018984:Epitopes, T-Lymphocyte; D005353:Fibronectins; D016056:Immunodominant Epitopes; D008565:Membrane Proteins; D010455:Peptides; D019089:Stem Cell Factor; C084881:flt3 ligand protein; D013926:Thrombopoietin