title: Rapid generation of NY-ESO-1-specific CD4+THELPER1 cells for adoptive T-cell therapy.
abstract: Tumor-associated antigens such as NY-ESO-1 are expressed in a variety of solid tumors but absent in mature healthy tissues with the exception of germline cells. The immune system anti-cancer attack is mediated by cell lysis or induction of growth arrest through paralysis of tumor cells, the latter of which can be achieved by tumor-specific CD4+, IFNγ-producing THelpertype 1 (TH1) cells. Translation of these immune-mediated mechanisms into clinical application has been limited by availability of immune effectors, as well as the need for complexin vitroprotocols and regulatory hurdles. Here, we report a procedure to generate cancer-testis antigen NY-ESO-1-targeting CD4+TH1 cellsin vitrofor cancer immunotherapy in the clinic. After in vitro sensitization by stimulating T cells with protein-spanning, overlapping peptide pools of NY-ESO-1 in combination with IL-7 and low dose IL-2, antigen-specific T cells were isolated using IFNγ capture technique and subsequently expanded with IL-2, IL-7 and IL-15. Large numbers of NY-ESO-1-specific CD4+T cells with a TH1 cytokine profile and lower numbers of cytokine-secreting CD8+T cells could be generated from healthy donors with a high specificity and expansion potential. Manufactured CD4+T cells showed strong specific TH1-responses with IFNγ+, TNFα+, IL-2+and induced cell cycle arrest and apoptosis in tumor cells. The protocol is GMP-grade and approved by the regulatory authorities. The tumor-antigen specific CD4+TH1 lymphocytes can be adoptively transferred as a T-cell therapy to boost anticancer immunity and this novel cancer treatment approach is applicable to both T cells from healthy allogeneic donors as well as to autologous T cells derived from cancer patients.
authors.affiliation: University Children's Hospital Tübingen ; Tübingen, Germany.
authors.forename: Simone
authors.initials: S
authors.lastname: Kayser
authors.affiliation: Department of Dermatology; University Hospital Tübingen ; Tübingen, Germany.
authors.forename: Cristina
authors.initials: C
authors.lastname: Boβ
authors.affiliation: University Children's Hospital Tübingen ; Tübingen, Germany.
authors.forename: Judith
authors.initials: J
authors.lastname: Feucht
authors.affiliation: University Children's Hospital Tübingen ; Tübingen, Germany.
authors.forename: Kai-Erik
authors.initials: KE
authors.lastname: Witte
authors.affiliation: University Children's Hospital Tübingen ; Tübingen, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Scheu
authors.affiliation: University Children's Hospital Tübingen ; Tübingen, Germany.
authors.forename: Hans-Jörg
authors.initials: HJ
authors.lastname: Bülow
authors.affiliation: University Children's Hospital Tübingen ; Tübingen, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Joachim
authors.affiliation: Interfaculty Institute for Cell Biology, Department of Immunology, University Tübingen , Tübingen , Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Stevanović
authors.affiliation: University Children's Hospital Tübingen ; Tübingen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Schumm
authors.affiliation: Department of Internal Medicine; University Hospital Tübingen ; Tübingen, Germany.
authors.forename: Susanne M
authors.initials: SM
authors.lastname: Rittig
authors.affiliation: University Children's Hospital Tübingen ; Tübingen, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Lang
authors.affiliation: Department of Dermatology; University Hospital Tübingen ; Tübingen, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Röcken
authors.affiliation: University Children's Hospital Tübingen ; Tübingen, Germany.
authors.forename: Rupert
authors.initials: R
authors.lastname: Handgretinger
authors.affiliation: Oncology and Stem Cell Transplantation; Dr. von Hauner'sches Kinderspital; Ludwig-Maximilians-University ; Munich, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Feuchtinger
keywords: ACT, adoptive T cell transfer; CD4+ THELPER1 cells; CTL, cytotoxic T lymphocyte; DC, dendritic cell; GMP, good manufacturing practice; NK, natural killer; TH1, THelper1; TIL, tumor-infiltrating lymphocyte; Treg, regulatory T cell; adoptive T-cell transfer; cell cycle arrest; immunotherapy; polyfunctional T cells