title: Human Neutrophil Elastase Induce Interleukin-10 Expression in Peripheral Blood Mononuclear Cells through Protein Kinase C Theta/Delta and Phospholipase Pathways.
abstract: OBJECTIVE
  Neutrophils have an important role in the rapid innate immune response, and the release or active secretion of elastase from neutrophils is linked to various inflammatory responses. Purpose of this study was to determine how the human neutrophil elastase affects the interleukin-10 (IL-10) response in peripheral blood mononuclear cells (PBMC).
  
  
  METHODS
  In this prospective study, changes in IL-10 messenger RNA (mRNA) and protein expression levels in monocytes derived from human PBMCs were investigated after stimulation with human neutrophil elastase (HNE). A set of inhibitors was used for examining the pathways for IL-10 production induced by HNE.
  
  
  RESULTS
  Reverse transcription polymerase chain reaction (RT-PCR) showed that stimulation with HNE upregulated IL-10 mRNA expression by monocytes, while the enzyme-linked immunosorbent assay (ELISA) revealed an increase of IL-10 protein level in the culture medium. A phospholipase C inhibitor (U73122) partially blunt- ed the induction of IL-10 mRNA expression by HNE, while IL-10 mRNA expression was significantly reduced by a protein kinase C (PKC) inhibitor (Rottlerin). A calcium chelator (3,4,5-trimethoxybenzoic acid 8-(diethylamino)octyl ester: TMB-8) inhibited the response of IL-10 mRNA to stimulation by HNE. In addition, pretreatment with a broad-spectrum PKC inhibitor (Ro-318425) partly blocked the response to HNE. Finally, an inhibitor of PKC theta/delta abolished the increased level of IL-10 mRNA expression.
  
  
  CONCLUSIONS
  These results indicate that HNE mainly upregulates IL-10 mRNA ex- pression and protein production in moncytes via a novel PKC theta/delta, although partially via the conventional PKC pathway.
authors: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors: Jin
authors: J
authors: Kawata
authors: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors: Rui
authors: R
authors: Yamaguchi
authors: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors: Takatoshi
authors: T
authors: Yamamoto
authors: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors: Yasuji
authors: Y
authors: Ishimaru
authors: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors: Arisa
authors: A
authors: Sakamoto
authors: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors: Manabu
authors: M
authors: Aoki
authors: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors: Masafumi
authors: M
authors: Kitano
authors: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors: Misako
authors: M
authors: Umehashi
authors: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors: Eiji
authors: E
authors: Hirose
authors: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors: Yasuo
authors: Y
authors: Yamaguchi
keywords: Interleukin-10; PBMC; Protein kinase C