IL4-10 fusion protein: a novel immunoregulatory drug combining activities of interleukin 4 and interleukin 10

C Steen-Louws; S A Y Hartgring; J Popov-Celeketic; A P Lopes; M B M de Smet; N Eijkelkamp; F P J G Lafeber; C E Hack; J A G van Roon

doi:10.1111/cei.13224

IL4-10 fusion protein: a novel immunoregulatory drug combining activities of interleukin 4 and interleukin 10

Clin Exp Immunol. 2019 Jan;195(1):1-9. doi: 10.1111/cei.13224. Epub 2018 Nov 11.

Authors

C Steen-Louws^{1

2}, S A Y Hartgring^{1

2}, J Popov-Celeketic², A P Lopes^{1

2}, M B M de Smet¹, N Eijkelkamp^{1

3}, F P J G Lafeber², C E Hack^{1

2}, J A G van Roon^{1

2}

Affiliations

¹ Laboratory of Translational Immunology, University Medical Center Utrecht, the Netherlands.
² Department of Rheumatology and Clinical Immunology, Utrecht, the Netherlands.
³ Laboratory of Neuroimmunology and Developmental Origins of Disease, Utrecht, the Netherlands.

Abstract

The objective of this study was to test the capacity of a newly developed fusion protein of interleukin 4 (IL-4) and IL-10 [IL4-10 fusion protein (FP)] to shift multiple pro-inflammatory pathways towards immune regulation, and to inhibit pro-inflammatory activity in arthritis models. The effects of IL4-10 FP in comparison with IL-4, IL-10 and IL-4 plus IL-10 on pro- and anti-inflammatory mediators, T cells and immunoglobulin (Ig) receptors in favour of immunoregulatory activity were studied. In addition, the capacity of IL4-10 FP to inhibit pro-inflammatory activity in ex-vivo and in-vivo arthritis models was investigated. IL4-10 FP robustly inhibited pro-inflammatory cytokine [IL-1β, tumour necrosis factor (TNF)-α, IL-6 and IL-8] production in whole blood cultures, mediated by both the IL-10 and the IL-4 moiety. IL4-10 fusion protein induced IL-1 receptor antagonist (IL-1RA) production and preserved soluble TNF receptor (sTNFR) levels, strongly increasing IL-1RA/IL-1β and sTNFR/TNF-α ratios. In addition, IL4-10 FP strongly inhibited T helper (Th) type 1 and 17 cytokine secretion, while maintaining FoxP3 expression and up-regulating Th2 activity. In addition, while largely leaving expression of activating Fc gamma receptor (FcγR)I, III and Fc epsilon receptor (FcεR) unaffected, it significantly shifted the FcγRIIa/FcγRIIb ratio in favour of the inhibitory FcγRIIb. Moreover, IL4-10 FP robustly inhibited secretion of pro-inflammatory cytokines by rheumatoid arthritis synovial tissue and suppressed experimental arthritis in mice, without inducing B cell hyperactivity. IL4-10 fusion protein is a novel drug, signalling cells to induce immunoregulatory activity that overcomes limitations of IL-4 and IL-10 stand-alone therapy, and therefore has therapeutic potential for inflammatory diseases such as rheumatoid arthritis.

Keywords: Th1/Th2 cells; arthritis; autoinflammatory diseases; cytokines; inflammation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Arthritis, Rheumatoid / chemically induced
Arthritis, Rheumatoid / immunology
Arthritis, Rheumatoid / therapy*
Cell Proliferation
Cells, Cultured
Disease Models, Animal
Female
Flow Cytometry
Humans
Immunomodulation
Immunotherapy / methods*
Inflammation / immunology
Inflammation / therapy*
Interleukin-10 / immunology*
Interleukin-4 / genetics
Interleukin-4 / therapeutic use*
Leukocytes, Mononuclear / immunology*
Lipopolysaccharides / immunology
Mice
Mice, Inbred BALB C
Proteoglycans
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / therapeutic use*
Synovial Membrane / metabolism
Synovial Membrane / pathology

Substances

Lipopolysaccharides
Proteoglycans
Recombinant Fusion Proteins
Interleukin-10
Interleukin-4