Antiinflammatory effect of retrovirally transfected interleukin‐10 on monosodium urate monohydrate crystal–induced acute inflammation in murine air pouches

Abstract

Objective. To investigate the role of interleukin-10 (IL-10) in the inflammatory response, the antiinflammatory effect of retrovirally transfected IL-10 was evaluated both in vitro and in vivo.Methods. A recombinant retrovirus containing the murine IL-10 gene was constructed using the pLXSN vector and was designated as LXSN-IL-10. Murine IL-10 was introduced into embryonic C57BL/6J fibroblast cells using LXSN-IL-10 to create C57-IL-10 cells. The effect of IL-10 in the culture supernatant of these cells was then evaluated by determining changes in the production of tumor necrosis factor ␣ (TNF␣), macrophage inflammatory protein 1␣ (MIP-1␣), and MIP-1␤ by macrophages. The antiinflammatory effect of C57-IL-10 cells was also investigated using an in vivo model of monosodium urate monohydrate (MSU) crystal-induced acute inflammation.Results. The IL-10 gene transcript and its product were detected by reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The level of IL-10 in the culture supernatant of C57-IL-10 cells was estimated to be 50 ng/ml. The culture supernatant of these cells exerted the biologic activity of IL-10, showing inhibition of TNF␣, MIP-1␣, and MIP-1␤ production by macrophages. Injection of C57-IL-10 cells into murine air pouches significantly inhibited MSU crystal-induced cellular infiltration (P < 0.01) and production of the mouse CXC chemokine KC (P < 0.05). These findings were consistent with the results obtained by the injection of recombinant human IL-10 into air pouches.Conclusion. In this murine air pouch model of MSU crystal-induced inflammation, IL-10 seemed to inhibit the recruitment of neutrophils at least partly by suppressing KC production. These findings seem to suggest that IL-10 gene therapy may be useful for inflammatory diseases.

Induction of triggering receptor expressed on myeloid cells 1 in murine resident peritoneal macrophages by monosodium urate monohydrate crystals

“…Maximum induction occurred at 4 hours, and expression subsequently declined 8 hours after stimulation. We previously demonstrated increased expression of IL-1␤, macrophage inflammatory protein 2, and KC in infiltrating cells in this air-pouch model (18,19). The time course of TREM-1 mRNA expression closely resembles gene expression of inflammatory cytokines and chemokines.…”

Signaling Events Involved in Macrophage Chemokine Expression in Response to Monosodium Urate Crystals

“…Administration of neutralizing Abs directed against IL-8 (10), MCP-1 (12), and growth-related oncogene ␣ (13) significantly reduced MSU crystal-induced leukocyte accumulation, suggesting an important role for these chemokines in acute gout. In parallel, the use of the murine air-pouch model has further confirmed that MSU crystal-mediated leukocyte recruitment correlates with chemokine (macrophage inflammatory protein (MIP)-1␣/CCL3, MIP-2/ CXCL2, MCP-1 and KC/CXCL1) and myeloid-related protein (S100A8, S100A9, and S100A8/A9) release (8,14,15). Consistent with these data, MSU crystals activate the expression of a number of cytokines and chemokines in various types of leukocytes.…”

“…In monocytes, MSU crystal administration results in the release of tumor necrosis factor-␣ (16), IL-1 (17), , and IL-8 (6). In macrophages, increased mRNA levels of tumor necrosis factor-␣, MIP-1␣, MIP-1␤, and KC (14) as well as of IL-␤ and MIP-2 (15) have been detected upon MSU crystal treatment. Similarly, neutrophils are known to respond to MSU crystals by secreting, among others, IL-1 (19), , and reactive oxygen species (21).…”

Preparation and <i>in Vitro</i>, <i>in Vivo</i> Characterization of Elastic Liposomes Encapsulating Cyclodextrin-Colchicine Complexes for Topical Delivery of Colchicine

“…18) The crystals were suspended in PBS at a concentration of 10 mg/ml. The crystals were sonicated to obtain rod-shaped crystals of uniform length (5 25 mm) and sterilized by autoclaving at 121°C for 30 min.…”