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

Abstract

SummaryThe 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.

Sialic Acid-Engineered IL4–10 Fusion Protein is Bioactive and Rapidly Cleared from the Circulation

“…Systemic administration of IL4-10 FP attenuates experimental arthritis in mice (8). Intrathecal administration of IL4-10 FP remarkably reduces pain in various mouse models of inflammatory and neuropathic pain, as well as intra-articular administration in dogs with experimental OA (8,9). These findings suggest a potential of IL4-10 FP for the treatment of diseases, such as inflammatory pain and OA, where local application would be preferred over systemic administration.…”

“…Several explanations for these results have been put forward including pharmacokinetic limitations, and lack of synergism with other cytokines when used as stand-alone drugs (5,6). IL4-10 FP, a fusion protein of IL-4 and IL-10, combines the effects of both cytokines in a single molecule (7,8), and overcomes some of the limitations of stand-alone IL-4 or IL-10. Previously, we published the immunoregulatory activity of IL4-10 fusion protein in vitro.…”

“…Previously, we published the immunoregulatory activity of IL4-10 fusion protein in vitro. In whole blood and peripheral blood mononuclear cell (PBMC) cultures IL4-10 FP strongly inhibits the production of LPS-induced pro-inflammatory cytokines such as IL-1β, TNF, IL-6, and IL-8, while having minimal, if any, effect on the production of IL-1RA and sTNFR (8). In addition, the therapeutic potential of IL4-10 FP was demonstrated in several disease models both ex vivo and in vivo.…”

“…In addition, the therapeutic potential of IL4-10 FP was demonstrated in several disease models both ex vivo and in vivo. The fusion protein inhibits pro-inflammatory cytokine production by rheumatoid arthritis (RA) and osteoarthritis (OA) synovial tissue and cartilage cultures (8,9). Systemic administration of IL4-10 FP attenuates experimental arthritis in mice (8).…”

“…The fusion protein inhibits pro-inflammatory cytokine production by rheumatoid arthritis (RA) and osteoarthritis (OA) synovial tissue and cartilage cultures (8,9). Systemic administration of IL4-10 FP attenuates experimental arthritis in mice (8). Intrathecal administration of IL4-10 FP remarkably reduces pain in various mouse models of inflammatory and neuropathic pain, as well as intra-articular administration in dogs with experimental OA (8,9).…”