title: Polyfunctional T-cell responses are disrupted by the ovarian cancer ascites environment and only partially restored by clinically relevant cytokines.
abstract: BACKGROUND
  Host T-cell responses are associated with favorable outcomes in epithelial ovarian cancer (EOC), but it remains unclear how best to promote these responses in patients. Toward this goal, we evaluated a panel of clinically relevant cytokines for the ability to enhance multiple T-cell effector functions (polyfunctionality) in the native tumor environment.
  
  
  RESULTS
  Experiments were performed with resident CD8+ and CD4+ T cells in bulk ascites cell preparations from high-grade serous EOC patients. T cells were stimulated with α-CD3 in the presence of 100% autologous ascites fluid with or without exogenous IL-2, IL-12, IL-18 or IL-21, alone or in combination. T-cell proliferation (Ki-67) and function (IFN-γ, TNF-α, IL-2, CCL4, and CD107a expression) were assessed by multi-parameter flow cytometry. In parallel, 27 cytokines were measured in culture supernatants. While ascites fluid had variable effects on CD8+ and CD4+ T-cell proliferation, it inhibited T-cell function in most patient samples, with CD107a, IFN-γ, and CCL4 showing the greatest inhibition. This was accompanied by reduced levels of IL-1β, IL-1ra, IL-9, IL-17, G-CSF, GM-CSF, Mip-1α, PDGF-bb, and bFGF in culture supernatants. T-cell proliferation was enhanced by exogenous IL-2, but other T-cell functions were largely unaffected by single cytokines. The combination of IL-2 with cytokines engaging complementary signaling pathways, in particular IL-12 and IL-18, enhanced expression of IFN-γ, TNF-α, and CCL4 in all patient samples by promoting polyfunctional T-cell responses. Despite this, other functional parameters generally remained inhibited.
  
  
  CONCLUSIONS
  The EOC ascites environment disrupts multiple T-cell functions, and exogenous cytokines engaging diverse signaling pathways only partially reverse these effects. Our results may explain the limited efficacy of cytokine therapies for EOC to date. Full restoration of T-cell function will require activation of signaling pathways beyond those engaged by IL-2, IL-12, IL-18, and IL-21.
authors.affiliation: Department of Microbiology and Biochemistry, University of Victoria, Victoria, British Columbia, Canada.
authors.forename: Eric
authors.initials: E
authors.lastname: Tran
authors.forename: Julie S
authors.initials: JS
authors.lastname: Nielsen
authors.forename: Darin A
authors.initials: DA
authors.lastname: Wick
authors.forename: Alvin V
authors.initials: AV
authors.lastname: Ng
authors.forename: Lisa D S
authors.initials: LD
authors.lastname: Johnson
authors.forename: Nancy J
authors.initials: NJ
authors.lastname: Nesslinger
authors.forename: Elissa
authors.initials: E
authors.lastname: McMurtrie
authors.forename: John R
authors.initials: JR
authors.lastname: Webb
authors.forename: Brad H
authors.initials: BH
authors.lastname: Nelson
mesh_terms: D000328:Adult; D000368:Aged; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000378:metabolism; D049109:Cell Proliferation; D016207:Cytokines / Q000378:metabolism*; D005260:Female; D006801:Humans; D018664:Interleukin-12 / Q000378:metabolism; D020382:Interleukin-18 / Q000378:metabolism; D007376:Interleukin-2 / Q000378:metabolism; D007378:Interleukins / Q000378:metabolism; D008875:Middle Aged; D010051:Ovarian Neoplasms / Q000097:blood / Q000378:metabolism*; D013601:T-Lymphocytes / Q000166:cytology*; D000098546:Interleukin-21
chemical_list: D016207:Cytokines; D020382:Interleukin-18; D007376:Interleukin-2; D007378:Interleukins; D018664:Interleukin-12; D000098546:Interleukin-21