title: Manipulating the tumor microenvironment ex vivo for enhanced expansion of tumor-infiltrating lymphocytes for adoptive cell therapy.
abstract: OBJECTIVE
  Cultured tumor fragments from melanoma metastases have been used for years as a source of tumor-infiltrating lymphocytes (TIL) for adoptive cell therapy (ACT). The expansion of tumor-reactive CD8(+) T cells with interleukin-2 (IL2) in these early cultures is critical in generating clinically active TIL infusion products, with a population of activated 4-1BB CD8(+) T cells recently found to constitute the majority of tumor-specific T cells.
  
  
  METHODS
  We used an agonistic anti-4-1BB antibody added during the initial tumor fragment cultures to provide in situ 4-1BB costimulation.
  
  
  RESULTS
  We found that addition of an agonistic anti-4-1BB antibody could activate 4-1BB signaling within early cultured tumor fragments and accelerated the rate of memory CD8(+) TIL outgrowth that were highly enriched for melanoma antigen specificity. This was associated with NFκB activation and the induction of T-cell survival and memory genes, as well as enhanced IL2 responsiveness, in the CD8(+) T cells in the fragments and emerging from the fragments. Early provision of 4-1BB costimulation also affected the dendritic cells (DC) by activating NFκB in DC and promoting their maturation inside the tumor fragments. Blocking HLA class I prevented the enhanced outgrowth of CD8(+) T cells with anti-4-1BB, suggesting that an ongoing HLA class I-mediated antigen presentation in early tumor fragment cultures plays a role in mediating tumor-specific CD8(+) TIL outgrowth.
  
  
  CONCLUSIONS
  Our results highlight a previously unrecognized concept in TIL ACT that the tumor microenvironment can be dynamically regulated in the initial tumor fragment cultures to regulate the types of T cells expanded and their functional characteristics.
authors.affiliation: Department of Melanoma Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas. The Immunology Program of the University of Texas Health Science Center, Graduate School of Biomedical Sciences, The University of Texas MD Anderson Cancer Center, Houston, Texas. jchacon@mail.med.upenn.edu laszlo.radvanyi@lionbio.com.
authors.forename: Jessica Ann
authors.initials: JA
authors.lastname: Chacon
authors.affiliation: Donald A. Adam Comprehensive Melanoma Research Center, H. Lee Moffitt Cancer Center, Tampa, Florida.
authors.forename: Amod A
authors.initials: AA
authors.lastname: Sarnaik
authors.affiliation: Department of Melanoma Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas. Lion Biotechnologies, Woodland Hills, Los Angeles, California.
authors.forename: Jie Qing
authors.initials: JQ
authors.lastname: Chen
authors.affiliation: Department of Melanoma Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas.
authors.forename: Caitlin
authors.initials: C
authors.lastname: Creasy
authors.affiliation: Department of Melanoma Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas.
authors.forename: Charuta
authors.initials: C
authors.lastname: Kale
authors.affiliation: Donald A. Adam Comprehensive Melanoma Research Center, H. Lee Moffitt Cancer Center, Tampa, Florida.
authors.forename: John
authors.initials: J
authors.lastname: Robinson
authors.affiliation: Donald A. Adam Comprehensive Melanoma Research Center, H. Lee Moffitt Cancer Center, Tampa, Florida.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Weber
authors.affiliation: Department of Melanoma Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas. The Immunology Program of the University of Texas Health Science Center, Graduate School of Biomedical Sciences, The University of Texas MD Anderson Cancer Center, Houston, Texas.
authors.forename: Patrick
authors.initials: P
authors.lastname: Hwu
authors.affiliation: Donald A. Adam Comprehensive Melanoma Research Center, H. Lee Moffitt Cancer Center, Tampa, Florida.
authors.forename: Shari
authors.initials: S
authors.lastname: Pilon-Thomas
authors.affiliation: Department of Melanoma Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas. The Immunology Program of the University of Texas Health Science Center, Graduate School of Biomedical Sciences, The University of Texas MD Anderson Cancer Center, Houston, Texas. Department of Immunology, H. Lee Moffitt Cancer Center, Tampa, Florida. Lion Biotechnologies, Woodland Hills, Los Angeles, California. jchacon@mail.med.upenn.edu laszlo.radvanyi@lionbio.com.
authors.forename: Laszlo
authors.initials: L
authors.lastname: Radvanyi
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D003713:Dendritic Cells / Q000187:drug effects / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping; D016219:Immunotherapy, Adoptive / Q000379:methods; D016246:Lymphocytes, Tumor-Infiltrating / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D008545:Melanoma / Q000276:immunology / Q000473:pathology / Q000628:therapy; D016328:NF-kappa B / Q000378:metabolism; D009369:Neoplasms / Q000235:genetics / Q000276:immunology* / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D010641:Phenotype; D015398:Signal Transduction / Q000187:drug effects; D059016:Tumor Microenvironment / Q000235:genetics / Q000276:immunology*; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000819:agonists / Q000235:genetics / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D016328:NF-kappa B; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9