A fusion protein of interleukin‐4 and interleukin‐10 protects against blood‐induced cartilage damage in vitro and in vivo

Abstract

Background Joint damage still causes significant morbidity in hemophilia. It results from synovial inflammation and direct cartilage-degenerating properties of blood components. Interleukin (IL)-4 and IL-10 have been shown to protect cartilage from blood-induced damage. Recently an IL4-10 fusion protein has been developed to combine the function of IL-4 and IL-10 and increase their bioavailability. Objectives In this study we evaluate whether this IL4-10 fusion protein protects against blood-induced joint damage. Methods In vitro, human cartilage explants were exposed to whole blood and simultaneously to a broad concentration range of the IL4-10 fusion protein. Effects on cartilage matrix turnover were compared with the individual cytokines. Moreover, the influence of the fusion protein and its individual components on IL-1β and IL-6 production was investigated. In hemophilia A mice, the effect of intra-articular treatment on synovitis and cartilage damage resulting from joint bleeding was evaluated by histochemistry. Results In vitro, the fusion protein prevented blood-induced cartilage damage in a dose-dependent manner, with equal effectiveness to the combination of the separate cytokines. In whole blood cultures 10 ng mL fusion protein completely blocked the production of IL-1β and IL-6 by monocytes/macrophages. In hemophilic mice, intra-articular injection of IL-4 and IL-10 did not influence synovitis or cartilage degeneration. In contrast, equimolar amounts of the fusion protein attenuated cartilage damage upon repeated joint bleeding, although synovial inflammation was hardly affected. Conclusions Overall, this study shows that the IL4-10 fusion protein prevents blood-induced cartilage damage in vitro and ameliorates cartilage degeneration upon joint bleeding in hemophilic mice.

Bleed volume of experimental knee haemarthrosis correlates with the subsequent degree of haemophilic arthropathy

“…On the other hand, cartilage degeneration was not observed. This aligns with previous findings and the fact that cartilage degeneration has been shown to be more pronounced in this model following two induced joint bleeds …”

“…This aligns with previous findings and the fact that cartilage degeneration has been shown to be more pronounced in this model following two induced joint bleeds. 16,35 Our findings substantially add to the value of this model, from which important knowledge on HA has been gained previously. 8 The observed variability in bleeding volume could reflect an exaggerated view of the clinical situation where "clinically evident joint bleeds" range from clinically unconfirmed joint bleeds based on the patients' experience all the way to the largest joint bleeds.…”

Resolution of inflammation-induced depression requires T lymphocytes and endogenous brain interleukin-10 signaling

“…Nasal administration of neutralizing anti-IL-10 antibody and recombinant IL-10 Thirty min before antibody or recombinant IL-10 fusion of human IL-10 and human IL-4 connected via a linker sequence to increase its stability [36][37][38], A concentration of 3 µl of hyaluronidase (total 100 U; Sigma-Aldrich) in PBS was delivered to each nostril [39]. c The tail-suspension test (TST) was performed in WT and Rag1 −/− mice at 24 h postinjection (n = 7 mice/group).…”

A Translational Study of TNF-Alpha Antagonists as an Adjunctive Therapy for Preventing Hemophilic Arthropathy

“…Pro-inflammatory cytokines have been reported to be involved in the pathogenesis of synovitis [25,28] as interactions within the cytokine network and other inflammatory mediators in the affected joint can create an inflammatory environment. Theoretically, an anti-cytokine approach can be used to protect against bleeding in damaged joints; however, limited studies have shown that intervention of the cytokine network may serve as a novel therapeutic direction [5,[29][30][31]; this includes our previous study with anti-IL-6 in hemophilia A mice [8].…”