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Dual biological functions of an interleukin‐1 receptor antagonist–interleukin‐10 fusion protein and its suppressive effects on joint inflammation
Author(s) -
Chang DehMing,
Shyue SongKun,
Liu ShaoHsiang,
Chen YenTeen,
Yeh ChiouYueh,
Lai JennHuang,
Lee HerngSheng,
Chen Ann
Publication year - 2004
Publication title -
immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.297
H-Index - 133
eISSN - 1365-2567
pISSN - 0019-2805
DOI - 10.1111/j.1365-2567.2004.01910.x
Subject(s) - fusion protein , interleukin 1 receptor antagonist , concanavalin a , splenocyte , interleukin , receptor antagonist , interleukin 2 , inflammation , microbiology and biotechnology , biology , receptor , arthritis , immunology , cytokine , antibody , recombinant dna , antagonist , in vitro , biochemistry , gene
Summary The aim of this study was to construct and purify a novel interleukin‐1 receptor antagonist (IL‐1ra)–interleukin‐10 (IL‐10) fusion protein and determine its biological function and anti‐inflammatory effects. The isolated cDNAs of two inhibitory cytokines (IL‐1ra, IL‐10) were used to construct a cDNA for the IL‐1ra–IL‐10 fusion protein. The expressed recombinant cytokines and fusion product were purified and their biological properties analysed. The anti‐IL‐1 effect was evaluated by using a thymocyte‐proliferation assay, and the IL‐10 effect was investigated by the inhibition of interferon‐γ (IFN‐γ) production from splenocytes. The clinical response and histological analyses were studied in an adjuvant arthritic rat model. The fusion protein was 38 000 molecular weight in size. Sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and immunoblotting demonstrated that the purified protein was recognized by both IL‐1ra and IL‐10 antibodies. The fusion protein significantly inhibited IL‐1‐mediated thymocyte proliferation and concanavalin A (ConA)‐primed IFN‐γ production from splenocytes. The fusion protein also suppressed joint swelling (paw circumference reduced from 5·0 ± 0·2 to 4·1 ± 0·1 cm; paw thickness ≈ 2 mm in difference) and synovial inflammation in adjuvant arthritis of rats. Our investigations indicate that this fusion protein effectively suppresses inflammatory arthritis and may initiate a trend for future clinical application to target multiple molecules at the same time.