title: Activation and propagation of tumor-infiltrating lymphocytes on clinical-grade designer artificial antigen-presenting cells for adoptive immunotherapy of melanoma.
abstract: OBJECTIVE
  Adoptive cell therapy with autologous tumor-infiltrating lymphocytes (TIL) is a therapy for metastatic melanoma with response rates of up to 50%. However, the generation of the TIL transfer product is challenging, requiring pooled allogeneic normal donor peripheral blood mononuclear cells (PBMC) used in vitro as "feeders" to support a rapid-expansion protocol. Here, we optimized a platform to propagate TIL to a clinical scale using K562 cells genetically modified to express costimulatory molecules such as CD86, CD137-ligand, and membrane-bound IL-15 to function as artificial antigen-presenting cells (aAPC) as an alternative to using PBMC feeders.
  
  
  METHODS
  We used aAPC or γ-irradiated PBMC feeders to propagate TIL and measured rates of expansion. The activation and differentiation state was evaluated by flow cytometry and differential gene expression analyses. Clonal diversity was assessed on the basis of the pattern of T-cell receptor usage. T-cell effector function was measured by evaluation of cytotoxic granule content and killing of target cells.
  
  
  RESULTS
  The aAPC propagated TIL at numbers equivalent to that found with PBMC feeders, whereas increasing the frequency of CD8 T-cell expansion with a comparable effector-memory phenotype. mRNA profiling revealed an upregulation of genes in the Wnt and stem-cell pathways with the aAPC. The aAPC platform did not skew clonal diversity, and CD8 T cells showed comparable antitumor function as those expanded with PBMC feeders.
  
  
  CONCLUSIONS
  TIL can be rapidly expanded with aAPC to clinical scale generating T cells with similar phenotypic and effector profiles as with PBMC feeders. These data support the clinical application of aAPC to manufacture TIL for the treatment of melanoma.
authors.affiliation: *Department of Melanoma Medical Oncology †Division of Pediatrics, MD Anderson Cancer Center, Houston, TX ‡Infectious Disease and Immunogenetics Section, Department of Transfusion Medicine, Clinical Center and trans-NIH Center for Human Immunology, National Institutes of Health, Bethesda, MD ∥Lion Biotechnologies, Woodland Hills, CA §Sidra Medical and Research Hospital, Doha, Qatar.
authors.forename: Marie-Andrée
authors.initials: MA
authors.lastname: Forget
authors.forename: Shruti
authors.initials: S
authors.lastname: Malu
authors.forename: Hui
authors.initials: H
authors.lastname: Liu
authors.forename: Christopher
authors.initials: C
authors.lastname: Toth
authors.forename: Sourindra
authors.initials: S
authors.lastname: Maiti
authors.forename: Charuta
authors.initials: C
authors.lastname: Kale
authors.forename: Cara
authors.initials: C
authors.lastname: Haymaker
authors.forename: Chantale
authors.initials: C
authors.lastname: Bernatchez
authors.forename: Helen
authors.initials: H
authors.lastname: Huls
authors.forename: Ena
authors.initials: E
authors.lastname: Wang
authors.forename: Francesco M
authors.initials: FM
authors.lastname: Marincola
authors.forename: Patrick
authors.initials: P
authors.lastname: Hwu
authors.forename: Laurence J N
authors.initials: LJ
authors.lastname: Cooper
authors.forename: Laszlo G
authors.initials: LG
authors.lastname: Radvanyi
mesh_terms: D000938:Antigen-Presenting Cells / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D006801:Humans; D016219:Immunotherapy, Adoptive; D020014:K562 Cells; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology*; D008545:Melanoma / Q000276:immunology*; D012878:Skin Neoplasms / Q000276:immunology*; D059467:Transcriptome