title: Analysis of nucleophosmin-anaplastic lymphoma kinase (NPM-ALK)-reactive CD8(+) T cell responses in children with NPM-ALK(+) anaplastic large cell lymphoma.
abstract: Cellular immune responses against the oncoantigen anaplastic lymphoma kinase (ALK) in patients with ALK-positive anaplastic large cell lymphoma (ALCL) have been detected using peptide-based approaches in individuals preselected for human leucocyte antigen (HLA)-A*02:01. In this study, we aimed to evaluate nucleophosmin (NPM)-ALK-specific CD8(+) T cell responses in ALCL patients ensuring endogenous peptide processing of ALK antigens and avoiding HLA preselection. We also examined the HLA class I restriction of ALK-specific CD8(+) T cells. Autologous dendritic cells (DCs) transfected with in-vitro-transcribed RNA (IVT-RNA) encoding NPM-ALK were used as antigen-presenting cells for T cell stimulation. Responder T lymphocytes were tested in interferon-gamma enzyme-linked immunospot (ELISPOT) assays with NPM-ALK-transfected autologous DCs as well as CV-1 in Origin with SV40 genes (COS-7) cells co-transfected with genes encoding the patients' HLA class I alleles and with NPM-ALK encoding cDNA to verify responses and define the HLA restrictions of specific T cell responses. NPM-ALK-specific CD8(+) T cell responses were detected in three of five ALK-positive ALCL patients tested between 1 and 13 years after diagnosis. The three patients had also maintained anti-ALK antibody responses. No reactivity was detected in samples from five healthy donors. The NPM-ALK-specific CD8(+) T cell responses were restricted by HLA-C-alleles (C*06:02 and C*12:02) in all three cases. This approach allowed for the detection of NPM-ALK-reactive T cells, irrespective of the individual HLA status, up to 9 years after ALCL diagnosis.
authors.affiliation: Department of Pediatric Hematology and Oncology.
authors.forename: V
authors.initials: V
authors.lastname: K Singh
authors.affiliation: Department of Pediatric Hematology and Oncology.
authors.forename: S
authors.initials: S
authors.lastname: Werner
authors.affiliation: Institute of Clinical Immunology and Transfusion Medicine, Justus-Liebig-University, Giessen, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Hackstein
authors.affiliation: Department of Internal Medicine III, University Medical Center, Johannes Gutenberg University, Mainz, Germany.
authors.forename: V
authors.initials: V
authors.lastname: Lennerz
authors.affiliation: Department of Pediatric Hematology and Oncology.
authors.forename: A
authors.initials: A
authors.lastname: Reiter
authors.affiliation: Department of Internal Medicine III, University Medical Center, Johannes Gutenberg University, Mainz, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Wölfel
authors.affiliation: Department of Pediatric Hematology and Oncology.
authors.forename: C
authors.initials: C
authors.lastname: Damm-Welk
authors.affiliation: Department of Pediatric Hematology and Oncology.
authors.forename: W
authors.initials: W
authors.lastname: Woessmann
mesh_terms: D000293:Adolescent; D000483:Alleles; D000818:Animals; D000906:Antibodies / Q000097:blood / Q000276:immunology; D000951:Antigens, Neoplasm / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D002648:Child; D002675:Child, Preschool; D015395:Histocompatibility Antigens Class I / Q000235:genetics / Q000276:immunology; D006801:Humans; D007223:Infant; D008213:Lymphocyte Activation / Q000276:immunology; D017728:Lymphoma, Large-Cell, Anaplastic / Q000175:diagnosis / Q000188:drug therapy / Q000276:immunology* / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000276:immunology* / Q000378:metabolism; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism
chemical_list: D000906:Antibodies; D000951:Antigens, Neoplasm; D015395:Histocompatibility Antigens Class I; C107023:p80(NPM-ALK) protein; D011505:Protein-Tyrosine Kinases
keywords: ALCL; CD8+ T cell; IFN-γ ELISPOT; NPM-ALK; immune response