Cargando…

Blocking the janus-activated kinase pathway reduces tumor necrosis factor alpha-induced interleukin-18 bioactivity by caspase-1 inhibition

INTRODUCTION: Our objective was to examine the role of the janus-activated kinase (JAK) pathway in the modulation of tumor necrosis factor-α (TNF)-induced-IL-18 bioactivity by reduction of caspase-1 function. METHODS: Caspase-1 expression in rheumatoid arthritis (RA) synovial fibroblasts treated wit...

Descripción completa

Detalles Bibliográficos
Autores principales: Marotte, Hubert, Tsou, Pei-Suen, Fedorova, Tatiana, Pinney, Adam J, Lewis, Benjamin, Koch, Alisa E
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4060193/
https://www.ncbi.nlm.nih.gov/pubmed/24762050
http://dx.doi.org/10.1186/ar4551
_version_ 1782321335768186880
author Marotte, Hubert
Tsou, Pei-Suen
Fedorova, Tatiana
Pinney, Adam J
Lewis, Benjamin
Koch, Alisa E
author_facet Marotte, Hubert
Tsou, Pei-Suen
Fedorova, Tatiana
Pinney, Adam J
Lewis, Benjamin
Koch, Alisa E
author_sort Marotte, Hubert
collection PubMed
description INTRODUCTION: Our objective was to examine the role of the janus-activated kinase (JAK) pathway in the modulation of tumor necrosis factor-α (TNF)-induced-IL-18 bioactivity by reduction of caspase-1 function. METHODS: Caspase-1 expression in rheumatoid arthritis (RA) synovial fibroblasts treated with TNF was assessed by qRT-PCR and Western blot. Interleukin (IL)-18 was assessed by enzyme-linked immunosorbent assay (ELISA) in cell lysates and conditioned media and detected by immunofluorescence (IF) staining in RA synovial fibroblasts. The critical pathways for TNF-induced caspase-1 expression were determined by using chemical inhibitors of signaling followed by TNF stimulation. IL-18 bioactivity was assessed using human myelomonocytic KG-1 cells. RESULTS: TNF induced RA synovial fibroblast caspase-1 expression at the protein level in a time-dependant manner (P < 0.05). Blocking the JAK pathway reduced TNF-induced-caspase-1 expression at the transcriptional and protein levels by approximately 60% and 40%, respectively (P < 0.05). Blocking the JAK pathway reduced TNF-induced-caspase-1 expression at the transcriptional, protein, and activity levels by approximately 60%, 40%, and 53%, respectively (P < 0.05). We then confirmed by IF that TNF-induced IL-18 and investigated roles of the ERK1/2 and JAK pathways. Blocking the JAK pathway, TNF induced intracytoplasmic granular IL-18 expression suggesting a defect of caspase-1. Finally, blocking the JAK pathway, we observed a reduction of IL-18 bioactivity by 52% in RA synovial fibroblasts (P < 0.05). CONCLUSIONS: These results provide a new way to regulate TNF-induced-IL-18 bioactivity by blocking capase-1. These data present a novel role for JAK inhibition in RA patients and emphasize JAK inhibition use as a new therapeutic option in RA management.
format Online
Article
Text
id pubmed-4060193
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-40601932014-06-17 Blocking the janus-activated kinase pathway reduces tumor necrosis factor alpha-induced interleukin-18 bioactivity by caspase-1 inhibition Marotte, Hubert Tsou, Pei-Suen Fedorova, Tatiana Pinney, Adam J Lewis, Benjamin Koch, Alisa E Arthritis Res Ther Research Article INTRODUCTION: Our objective was to examine the role of the janus-activated kinase (JAK) pathway in the modulation of tumor necrosis factor-α (TNF)-induced-IL-18 bioactivity by reduction of caspase-1 function. METHODS: Caspase-1 expression in rheumatoid arthritis (RA) synovial fibroblasts treated with TNF was assessed by qRT-PCR and Western blot. Interleukin (IL)-18 was assessed by enzyme-linked immunosorbent assay (ELISA) in cell lysates and conditioned media and detected by immunofluorescence (IF) staining in RA synovial fibroblasts. The critical pathways for TNF-induced caspase-1 expression were determined by using chemical inhibitors of signaling followed by TNF stimulation. IL-18 bioactivity was assessed using human myelomonocytic KG-1 cells. RESULTS: TNF induced RA synovial fibroblast caspase-1 expression at the protein level in a time-dependant manner (P < 0.05). Blocking the JAK pathway reduced TNF-induced-caspase-1 expression at the transcriptional and protein levels by approximately 60% and 40%, respectively (P < 0.05). Blocking the JAK pathway reduced TNF-induced-caspase-1 expression at the transcriptional, protein, and activity levels by approximately 60%, 40%, and 53%, respectively (P < 0.05). We then confirmed by IF that TNF-induced IL-18 and investigated roles of the ERK1/2 and JAK pathways. Blocking the JAK pathway, TNF induced intracytoplasmic granular IL-18 expression suggesting a defect of caspase-1. Finally, blocking the JAK pathway, we observed a reduction of IL-18 bioactivity by 52% in RA synovial fibroblasts (P < 0.05). CONCLUSIONS: These results provide a new way to regulate TNF-induced-IL-18 bioactivity by blocking capase-1. These data present a novel role for JAK inhibition in RA patients and emphasize JAK inhibition use as a new therapeutic option in RA management. BioMed Central 2014 2014-04-24 /pmc/articles/PMC4060193/ /pubmed/24762050 http://dx.doi.org/10.1186/ar4551 Text en Copyright © 2014 Marotte et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Marotte, Hubert
Tsou, Pei-Suen
Fedorova, Tatiana
Pinney, Adam J
Lewis, Benjamin
Koch, Alisa E
Blocking the janus-activated kinase pathway reduces tumor necrosis factor alpha-induced interleukin-18 bioactivity by caspase-1 inhibition
title Blocking the janus-activated kinase pathway reduces tumor necrosis factor alpha-induced interleukin-18 bioactivity by caspase-1 inhibition
title_full Blocking the janus-activated kinase pathway reduces tumor necrosis factor alpha-induced interleukin-18 bioactivity by caspase-1 inhibition
title_fullStr Blocking the janus-activated kinase pathway reduces tumor necrosis factor alpha-induced interleukin-18 bioactivity by caspase-1 inhibition
title_full_unstemmed Blocking the janus-activated kinase pathway reduces tumor necrosis factor alpha-induced interleukin-18 bioactivity by caspase-1 inhibition
title_short Blocking the janus-activated kinase pathway reduces tumor necrosis factor alpha-induced interleukin-18 bioactivity by caspase-1 inhibition
title_sort blocking the janus-activated kinase pathway reduces tumor necrosis factor alpha-induced interleukin-18 bioactivity by caspase-1 inhibition
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4060193/
https://www.ncbi.nlm.nih.gov/pubmed/24762050
http://dx.doi.org/10.1186/ar4551
work_keys_str_mv AT marottehubert blockingthejanusactivatedkinasepathwayreducestumornecrosisfactoralphainducedinterleukin18bioactivitybycaspase1inhibition
AT tsoupeisuen blockingthejanusactivatedkinasepathwayreducestumornecrosisfactoralphainducedinterleukin18bioactivitybycaspase1inhibition
AT fedorovatatiana blockingthejanusactivatedkinasepathwayreducestumornecrosisfactoralphainducedinterleukin18bioactivitybycaspase1inhibition
AT pinneyadamj blockingthejanusactivatedkinasepathwayreducestumornecrosisfactoralphainducedinterleukin18bioactivitybycaspase1inhibition
AT lewisbenjamin blockingthejanusactivatedkinasepathwayreducestumornecrosisfactoralphainducedinterleukin18bioactivitybycaspase1inhibition
AT kochalisae blockingthejanusactivatedkinasepathwayreducestumornecrosisfactoralphainducedinterleukin18bioactivitybycaspase1inhibition