title: CD4 T-cell cytokines synergize to induce proliferation of malignant and nonmalignant innate intraepithelial lymphocytes.
abstract: Refractory celiac disease type II (RCDII) is a severe complication of celiac disease (CD) characterized by the presence of an enlarged clonal population of innate intraepithelial lymphocytes (IELs) lacking classical B-, T-, and natural killer (NK)-cell lineage markers (Lin-IELs) in the duodenum. In ∼50% of patients with RCDII, these Lin-IELs develop into a lymphoma for which no effective treatment is available. Current evidence indicates that the survival and expansion of these malignant Lin-IELs is driven by epithelial cell-derived IL-15. Like CD, RCDII is strongly associated with HLA-DQ2, suggesting the involvement of HLA-DQ2-restricted gluten-specific CD4+T cells. We now show that gluten-specific CD4+T cells isolated from CD duodenal biopsy specimens produce cytokines able to trigger proliferation of malignant Lin-IEL lines as powerfully as IL-15. Furthermore, we identify TNF, IL-2, and IL-21 as CD4+T-cell cytokines that synergistically mediate this effect. Like IL-15, these cytokines were found to increase the phosphorylation of STAT5 and Akt and transcription of antiapoptotic mediator bcl-xLSeveral small-molecule inhibitors targeting the JAK/STAT pathway blocked proliferation elicited by IL-2 and IL-15, but only an inhibitor targeting the PI3K/Akt/mTOR pathway blocked proliferation induced by IL-15 as well as the CD4+T-cell cytokines. Confirming and extending these findings, TNF, IL-2, and IL-21 also synergistically triggered the proliferation of freshly isolated Lin-IELs and CD3-CD56+IELs (NK-IELs) from RCDII as well as non-RCDII duodenal biopsy specimens. These data provide evidence implicating CD4+T-cell cytokines in the pathogenesis of RCDII. More broadly, they suggest that adaptive immune responses can contribute to innate IEL activation during mucosal inflammation.
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden 2333 ZA, The Netherlands.
authors.forename: Yvonne M C
authors.initials: YM
authors.lastname: Kooy-Winkelaar
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden 2333 ZA, The Netherlands.
authors.forename: Dagmar
authors.initials: D
authors.lastname: Bouwer
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden 2333 ZA, The Netherlands.
authors.forename: George M C
authors.initials: GM
authors.lastname: Janssen
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden 2333 ZA, The Netherlands.
authors.forename: Allan
authors.initials: A
authors.lastname: Thompson
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden 2333 ZA, The Netherlands.
authors.forename: Martijn H
authors.initials: MH
authors.lastname: Brugman
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden 2333 ZA, The Netherlands.
authors.forename: Frederike
authors.initials: F
authors.lastname: Schmitz
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden 2333 ZA, The Netherlands.
authors.forename: Arnoud H
authors.initials: AH
authors.lastname: de Ru
authors.affiliation: Department of Gastroenterology and Hepatology, VU University Medical Center, Amsterdam 1081 HZ, The Netherlands.
authors.forename: Tom
authors.initials: T
authors.lastname: van Gils
authors.affiliation: Department of Gastroenterology and Hepatology, VU University Medical Center, Amsterdam 1081 HZ, The Netherlands.
authors.forename: Gerd
authors.initials: G
authors.lastname: Bouma
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden 2333 ZA, The Netherlands; vanrood@europdonor.nl J.van_Bergen@lumc.nl.
authors.forename: Jon J
authors.initials: JJ
authors.lastname: van Rood
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden 2333 ZA, The Netherlands.
authors.forename: Peter A
authors.initials: PA
authors.lastname: van Veelen
authors.affiliation: Department of Pediatrics, Leiden University Medical Center, Leiden 2333 ZA, The Netherlands.
authors.forename: M Luisa
authors.initials: ML
authors.lastname: Mearin
authors.affiliation: Department of Gastroenterology and Hepatology, VU University Medical Center, Amsterdam 1081 HZ, The Netherlands.
authors.forename: Chris J
authors.initials: CJ
authors.lastname: Mulder
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden 2333 ZA, The Netherlands.
authors.forename: Frits
authors.initials: F
authors.lastname: Koning
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden 2333 ZA, The Netherlands; vanrood@europdonor.nl J.van_Bergen@lumc.nl.
authors.forename: Jeroen
authors.initials: J
authors.lastname: van Bergen
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism*; D002446:Celiac Disease / Q000235:genetics / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects* / Q000235:genetics; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D004357:Drug Synergism; D004386:Duodenum / Q000378:metabolism; D006801:Humans; D019409:Interleukin-15 / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D007376:Interleukin-2 / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D007378:Interleukins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D000075942:Intraepithelial Lymphocytes / Q000187:drug effects* / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D059467:Transcriptome / Q000187:drug effects / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D000098546:Interleukin-21
chemical_list: D016207:Cytokines; D019409:Interleukin-15; D007376:Interleukin-2; D007378:Interleukins; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D000098546:Interleukin-21
keywords: celiac disease; enteropathy-associated T-cell lymphoma; intraepithelial lymphocyte; refractory celiac disease; small-molecule inhibitor