IL-13 signaling via IL-13Rα(2) triggers TGF-β(1)-dependent allograft fibrosis

BACKGROUND: Allograft fibrosis still remains a critical problem in transplantation, including heart transplantation. The IL-13/TGF-β(1) interaction has previously been identified as a key pathway orchestrating fibrosis in different inflammatory immune disorders. Here we investigate if this pathway is also responsible for allograft fibrosis and if interference with the IL-13/TGF-β(1) interaction prevents allograft fibrosis. METHODS: FVB or control DBA/1 donor hearts were transplanted heterotopically into DBA/1 recipient mice and hearts were explanted at day 60 and 100 post-transplantation. Cardiac tissue was examined by Masson’s trichrome staining and immunohistochemistry for CD4, CD8, CD11b, IL-13, Fas ligand, matrix metalloproteinase (MMP)-1, MMP-13, β2-microglobulin, and Gremlin-1. Graft-infiltrating cells were isolated and analyzed by flow cytometry. IL-13 and TGF-β(1) levels were determined by enzyme-linked immunosorbent assay (ELISA) and the amount of collagen was quantified using a Sircol assay; IL-13Rα(2) expression was detected by Western blotting. In some experiments IL-13/ TGF-β(1) signaling was blocked with specific IL-13Rα(2) siRNA. Additionally, a PCR array of RNA isolated from the allografts was performed to analyze expression of multiple genes involved in fibrosis. RESULTS: Both groups survived long-term (>100 days). The allogeneic grafts were infiltrated by significantly increased numbers of CD4(+) (P <0.0001), CD8(+) (P <0.0001), and CD11b(+) cells (P = 0.0065) by day 100. Furthermore, elevated IL-13 levels (P = 0.0003) and numbers of infiltrating IL-13(+) cells (P = 0.0037), together with an expression of IL-13Rα(2), were detected only within allografts. The expression of IL-13 and IL-13Rα(2) resulted in significantly increased TGF-β(1) levels (P <0.0001), higher numbers of CD11b(high)Gr1(intermediate)TGF-β(1)(+) cells, and elevated cardiac collagen deposition (P = 0.0094). The allograft fibrosis found in these experiments was accompanied by upregulation of multiple profibrotic genes, which was confirmed by immunohistochemical stainings of allograft tissue. Blockage of the IL-13/TGF-β(1) interaction by IL-13Rα(2) siRNA led to lower numbers of CD11b(high)Gr1(intermediate)TGF-β(1)(+), CD4(+), CD8(+), and CD11b(+) cells, and prevented collagen deposition (P = 0.0018) within these allografts. CONCLUSIONS: IL-13 signaling via IL-13Rα(2) induces TGF-β(1) and causes allograft fibrosis in a murine model of chronic transplant rejection. Blockage of this IL-13/TGF-β(1) interaction by IL-13Rα(2) siRNA prevents cardiac allograft fibrosis. Thus, IL-13Rα(2) may be exploitable as a future target to reduce allograft fibrosis in organ transplantation.