Human Neutrophil Elastase Induce Interleukin-10 Expression in Peripheral Blood Mononuclear Cells through Protein Kinase C Theta/Delta and Phospholipase Pathways

(Pages: 692-700)
Jin Kawata, Ph.D, Rui Yamaguchi, Ph.D., Takatoshi Yamamoto, M.Sc., Yasuji Ishimaru, M.D., Ph.D., Arisa Sakamoto, M.Sc., Manabu Aoki, Ph.D., Masafumi Kitano, M.Sc., Misako Umehashi, Ph.D., Eiji Hirose, M.Sc., Yasuo Yamaguchi, M.D., Ph.D., *
Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan
Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan
*Corresponding Address: Graduate School of Medical Science Kumamoto Health Science University Kitaku Izumi-machi 325 Kumamoto 861-5598 Japan Email:yamaguti@kumamoto-hsu.ac.jp
Any use, distribution, reproduction or abstract of this publication in any medium, with the exception of commercial purposes, is permitted provided the original work is properly cited This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Kawata Jin, Yamaguchi Rui, Yamamoto Takatoshi, Ishimaru Yasuji, Sakamoto Arisa, Aoki Manabu, Kitano Masafumi, Umehashi Misako, Hirose Eiji, Yamaguchi Yasuo. Human Neutrophil Elastase Induce Interleukin-10 Expression in Peripheral Blood Mononuclear Cells through Protein Kinase C Theta/Delta and Phospholipase Pathways. Cell J. 2016; 17(4): 692-700.

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).

Materials and 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.

Conclusion

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.