title: Clinical grade generation of hexon-specific T cells for adoptive T-cell transfer as a treatment of adenovirus infection after allogeneic stem cell transplantation.
abstract: Adenovirus infection after allogeneic hematopoietic stem cell transplantation is still causing significant morbidity and mortality, especially in children. It has been demonstrated that a sufficient host T-cell response is essential to clear the virus. Adoptive transfer of specific T-cell immunity from the donor to the recipient has become a new treatment option for patients with systemic adenoviral infection who lack specific T-cell responses. The adenoviral hexon protein was shown to be an immunodominant T-cell target. We describe here a Good Manufacturing Practice-compatible generation of hexon-specific T cells developed by isolating interferon-gamma-secreting T cells after stimulation of mononuclear cells ex vivo with hexon protein. Phenotypical and functional characterization of the generated, specific T-cell product resulted in a mixed population of CD4 and CD8-positive T cells with an intermediate effector memory phenotype. Isolated hexon-specific T cells showed high expansion potential in vitro and specific cytotoxicity. T-cell lines, directed against type 5 hexon protein showed good crossreactivity against viral strains from other adenovirus species. The availability for isolation of hexon-specific T cells among 76 hematopoietic stem cell transplantation donors showed in > 72% a sufficient T-cell response (0.05% of T cells). In conclusion, Good Manufacturing Practice-grade selection of adenovirus-specific T cells for adoptive immunotherapy by hexon-induced secretion of interferon-gamma has been established. Adoptive T-cell transfer could potentially restore T-cell immunity against adenovirus after allogeneic stem cell transplantation.
authors.affiliation: Department of Pediatric Hematology/Oncology, University Children's Hospital, Eberhard-Karls University, Hoppe-Seyler Strasse 1, D-72076 Tuebingen, Germany. tobias.feuchtinger@med.uni-tuebingen.de
authors.forename: Tobias
authors.initials: T
authors.lastname: Feuchtinger
authors.forename: Celine
authors.initials: C
authors.lastname: Richard
authors.forename: Stefanie
authors.initials: S
authors.lastname: Joachim
authors.forename: Michael H
authors.initials: MH
authors.lastname: Scheible
authors.forename: Michael
authors.initials: M
authors.lastname: Schumm
authors.forename: Klaus
authors.initials: K
authors.lastname: Hamprecht
authors.forename: David
authors.initials: D
authors.lastname: Martin
authors.forename: Gerhard
authors.initials: G
authors.lastname: Jahn
authors.forename: Rupert
authors.initials: R
authors.lastname: Handgretinger
authors.forename: Peter
authors.initials: P
authors.lastname: Lang
mesh_terms: D000256:Adenoviridae / Q000276:immunology; D000258:Adenovirus Infections, Human / Q000209:etiology / Q000628:therapy*; D019264:Adoptive Transfer / Q000379:methods*; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D036022:Capsid Proteins / Q000276:immunology*; D002454:Cell Differentiation / Q000276:immunology; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D005347:Fibroblasts / Q000378:metabolism / Q000821:virology; D005434:Flow Cytometry; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects / Q000379:methods*; D006801:Humans; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D016131:Lymphocyte Subsets / Q000737:chemistry / Q000276:immunology; D037182:T-Cell Antigen Receptor Specificity / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D014184:Transplantation, Homologous
chemical_list: D036022:Capsid Proteins; C018315:hexon capsid protein, Adenovirus; D007371:Interferon-gamma