title: Identification and in vitro expansion of CD4+ and CD8+ T cells specific for human neutrophil elastase.
abstract: Human neutrophil elastase (HNE) and proteinase 3 (PRO3) are myeloid tissue-restricted serine proteases, aberrantly expressed by myeloid leukemia cells. PRO3 and HNE share the PR1 peptide sequence that induces HLA-A*0201-restricted cytotoxic T cells (CTLs) with antileukemia reactivity. We studied the entire HNE protein for its ability to induce CTLs. In an 18-hour culture, HNE-loaded monocytes stimulated significant intracellular interferon gamma (IFN-gamma) production by CD4+ and CD8+ T cells in 12 of 20 and 8 of 20 healthy individuals, respectively. Lymphocytes from 2 HNE responders were pulsed weekly for 4 weeks to generate HNE-specific CTLs. One of 2 HLA-A*0201-negative individuals inhibited the colony formation of HLA-identical chronic myelogenous leukemia progenitor cells (73% inhibition at 50:1 effector-target [E/T] ratio), indicating that peptides other than PR1 can induce leukemia-reactive CTLs. Repetitive stimulations with HNE in 2 of 5 HLA-A*0201+ individuals increased PR1 tetramer-positive CD8+ T-cell frequencies from 0.1% to 0.29% and 0.02% to 0.55%, respectively. These CTLs recognized PR1 peptide or killed HNE-loaded targets. These results indicate that exogenously processed HNE is a source of PR1 peptide as well as other peptide sequences capable of inducing leukemia-specific CD8+ and CD4+ T cells. HNE could, therefore, be used in an HLA-unrestricted manner to induce leukemia-reactive CTLs for adoptive immunotherapy.
authors.affiliation: Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Fujiwara
authors.forename: Frank
authors.initials: F
authors.lastname: El Ouriaghli
authors.forename: Matthias
authors.initials: M
authors.lastname: Grube
authors.forename: David A
authors.initials: DA
authors.lastname: Price
authors.forename: Katayoun
authors.initials: K
authors.lastname: Rezvani
authors.forename: Emma
authors.initials: E
authors.lastname: Gostick
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Sconocchia
authors.forename: Jos
authors.initials: J
authors.lastname: Melenhorst
authors.forename: Nancy
authors.initials: N
authors.lastname: Hensel
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Douek
authors.forename: A John
authors.initials: AJ
authors.lastname: Barrett
mesh_terms: D000595:Amino Acid Sequence; D000941:Antigens / Q000008:administration & dosage / Q000235:genetics; D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D015870:Gene Expression; D015234:HLA-A Antigens; D015789:HLA-A2 Antigen; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000096:biosynthesis; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000201:enzymology / Q000276:immunology / Q000628:therapy; D019272:Leukocyte Elastase / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D053146:Myeloblastin; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012334:RNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000235:genetics / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D000941:Antigens; D015234:HLA-A Antigens; C435939:HLA-A*02:01 antigen; D015789:HLA-A2 Antigen; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D007371:Interferon-gamma; D012697:Serine Endopeptidases; D019272:Leukocyte Elastase; D053146:Myeloblastin