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Rheumatoid arthritis synovial fibroblasts produce a soluble form of the interleukin-7 receptor in response to pro-inflammatory cytokines.

Badot V, Durez P, Van den Eynde BJ, Nzeusseu-Toukap A, Houssiau FA, Lauwerys BR - J. Cell. Mol. Med. (2011)

Bottom Line: We found that exposure of RA FLS to pro-inflammatory cytokines (TNF-α, IL-1β and combinations of TNF-α and IL-1β or TNF-α and IL-17) induces sIL-7R secretion.In DMARD-resistant patients, high sIL-7R serum concentrations are strongly associated with poor response to TNF-blockade.In conclusion, sIL-7R is induced by pro-inflammatory cytokines in RA FLS. sIL-7R could qualify as a new biomarker of response to therapy in RA.

View Article: PubMed Central - PubMed

Affiliation: Rheumatology Department, Cliniques Universitaires Saint-Luc, Université catholique de Louvain, Brussels, Belgium.

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Pro-inflammatory cytokines stimulate sIL-7R production by FLS. (A) IL-7R (open bars) and sIL-7R (closed bars) real-time qPCR studies carried out on FLS stimulated with the indicated cytokines. Results are expressed as mean fold changes in IL-7R and sIL-7R gene expression (±S.E.M.) over unstimulated FLS, obtained from two to five different experiments each. (B) Flow cytometric evaluation of IL-7R expression by PBMC and FLS. Cells were incubated with a PE-conjugated IL-7R antibody (red) or a PE-conjugated isotype control (blue). Autofluorescence of the cells is depicted in green. Graphs are representative of three different experiments. Similar results were obtained using FLS stimulated with pro-inflammatory cytokines, alone or in combination. (C) sIL-7R measurements were performed by sandwich-ELISA in supernatants of FLS cultures stimulated with the indicated cytokines. Results are expressed as mean optical density (O.D.) units ×1000 (after subtraction of the baseline O.D.) ±S.E.M. obtained from three different experiments. (D) Effect of IL-7 and sIL-7R-Fc fusion protein on proliferation of synovial CD4 T cells cultured in the presence of autologous FLS, autologous serum and IL-2. Results are expressed as mean cpm (±S.E.M.) obtained from two different experiments. *P < 0.05; **P < 0.005; ***P < 0.0005 versus unstimulated cells.
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fig02: Pro-inflammatory cytokines stimulate sIL-7R production by FLS. (A) IL-7R (open bars) and sIL-7R (closed bars) real-time qPCR studies carried out on FLS stimulated with the indicated cytokines. Results are expressed as mean fold changes in IL-7R and sIL-7R gene expression (±S.E.M.) over unstimulated FLS, obtained from two to five different experiments each. (B) Flow cytometric evaluation of IL-7R expression by PBMC and FLS. Cells were incubated with a PE-conjugated IL-7R antibody (red) or a PE-conjugated isotype control (blue). Autofluorescence of the cells is depicted in green. Graphs are representative of three different experiments. Similar results were obtained using FLS stimulated with pro-inflammatory cytokines, alone or in combination. (C) sIL-7R measurements were performed by sandwich-ELISA in supernatants of FLS cultures stimulated with the indicated cytokines. Results are expressed as mean optical density (O.D.) units ×1000 (after subtraction of the baseline O.D.) ±S.E.M. obtained from three different experiments. (D) Effect of IL-7 and sIL-7R-Fc fusion protein on proliferation of synovial CD4 T cells cultured in the presence of autologous FLS, autologous serum and IL-2. Results are expressed as mean cpm (±S.E.M.) obtained from two different experiments. *P < 0.05; **P < 0.005; ***P < 0.0005 versus unstimulated cells.

Mentions: Because sIL-7R transcripts result from an alternative splicing of IL-7R mRNAs, we wondered whether expression of both IL-7R isoforms is similarly regulated in FLS by the addition of pro-inflammatory cytokines known to play a role in the pathogenesis of RA. We found that mRNA levels of both the membrane-bound and soluble forms of IL-7R are up-regulated by the addition of TNF-α, IL-1β and combinations of TNF-α and IL-1β or TNF-α and IL-17 (Fig. 2A). Flow cytometry experiments were, however, unable to detect any cell surface expression of the membrane-bound IL-7R on FLS under any of these conditions (Fig. 2B). By contrast, ELISA experiments performed on culture supernatants indicated that sIL-7R secretion is induced in FLS by TNF-α, IL-1β and the combination of both cytokines (Fig. 2C). Interestingly, sIL-7R gene expression is negative in activated CD8 T and B cells but slightly positive in activated CD4 T cells (Fig. 1A). Taken together, these results indicate that sIL-7R is a marker of fibroblast and, to a lesser extent, CD4 T-cell activation.


Rheumatoid arthritis synovial fibroblasts produce a soluble form of the interleukin-7 receptor in response to pro-inflammatory cytokines.

Badot V, Durez P, Van den Eynde BJ, Nzeusseu-Toukap A, Houssiau FA, Lauwerys BR - J. Cell. Mol. Med. (2011)

Pro-inflammatory cytokines stimulate sIL-7R production by FLS. (A) IL-7R (open bars) and sIL-7R (closed bars) real-time qPCR studies carried out on FLS stimulated with the indicated cytokines. Results are expressed as mean fold changes in IL-7R and sIL-7R gene expression (±S.E.M.) over unstimulated FLS, obtained from two to five different experiments each. (B) Flow cytometric evaluation of IL-7R expression by PBMC and FLS. Cells were incubated with a PE-conjugated IL-7R antibody (red) or a PE-conjugated isotype control (blue). Autofluorescence of the cells is depicted in green. Graphs are representative of three different experiments. Similar results were obtained using FLS stimulated with pro-inflammatory cytokines, alone or in combination. (C) sIL-7R measurements were performed by sandwich-ELISA in supernatants of FLS cultures stimulated with the indicated cytokines. Results are expressed as mean optical density (O.D.) units ×1000 (after subtraction of the baseline O.D.) ±S.E.M. obtained from three different experiments. (D) Effect of IL-7 and sIL-7R-Fc fusion protein on proliferation of synovial CD4 T cells cultured in the presence of autologous FLS, autologous serum and IL-2. Results are expressed as mean cpm (±S.E.M.) obtained from two different experiments. *P < 0.05; **P < 0.005; ***P < 0.0005 versus unstimulated cells.
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fig02: Pro-inflammatory cytokines stimulate sIL-7R production by FLS. (A) IL-7R (open bars) and sIL-7R (closed bars) real-time qPCR studies carried out on FLS stimulated with the indicated cytokines. Results are expressed as mean fold changes in IL-7R and sIL-7R gene expression (±S.E.M.) over unstimulated FLS, obtained from two to five different experiments each. (B) Flow cytometric evaluation of IL-7R expression by PBMC and FLS. Cells were incubated with a PE-conjugated IL-7R antibody (red) or a PE-conjugated isotype control (blue). Autofluorescence of the cells is depicted in green. Graphs are representative of three different experiments. Similar results were obtained using FLS stimulated with pro-inflammatory cytokines, alone or in combination. (C) sIL-7R measurements were performed by sandwich-ELISA in supernatants of FLS cultures stimulated with the indicated cytokines. Results are expressed as mean optical density (O.D.) units ×1000 (after subtraction of the baseline O.D.) ±S.E.M. obtained from three different experiments. (D) Effect of IL-7 and sIL-7R-Fc fusion protein on proliferation of synovial CD4 T cells cultured in the presence of autologous FLS, autologous serum and IL-2. Results are expressed as mean cpm (±S.E.M.) obtained from two different experiments. *P < 0.05; **P < 0.005; ***P < 0.0005 versus unstimulated cells.
Mentions: Because sIL-7R transcripts result from an alternative splicing of IL-7R mRNAs, we wondered whether expression of both IL-7R isoforms is similarly regulated in FLS by the addition of pro-inflammatory cytokines known to play a role in the pathogenesis of RA. We found that mRNA levels of both the membrane-bound and soluble forms of IL-7R are up-regulated by the addition of TNF-α, IL-1β and combinations of TNF-α and IL-1β or TNF-α and IL-17 (Fig. 2A). Flow cytometry experiments were, however, unable to detect any cell surface expression of the membrane-bound IL-7R on FLS under any of these conditions (Fig. 2B). By contrast, ELISA experiments performed on culture supernatants indicated that sIL-7R secretion is induced in FLS by TNF-α, IL-1β and the combination of both cytokines (Fig. 2C). Interestingly, sIL-7R gene expression is negative in activated CD8 T and B cells but slightly positive in activated CD4 T cells (Fig. 1A). Taken together, these results indicate that sIL-7R is a marker of fibroblast and, to a lesser extent, CD4 T-cell activation.

Bottom Line: We found that exposure of RA FLS to pro-inflammatory cytokines (TNF-α, IL-1β and combinations of TNF-α and IL-1β or TNF-α and IL-17) induces sIL-7R secretion.In DMARD-resistant patients, high sIL-7R serum concentrations are strongly associated with poor response to TNF-blockade.In conclusion, sIL-7R is induced by pro-inflammatory cytokines in RA FLS. sIL-7R could qualify as a new biomarker of response to therapy in RA.

View Article: PubMed Central - PubMed

Affiliation: Rheumatology Department, Cliniques Universitaires Saint-Luc, Université catholique de Louvain, Brussels, Belgium.

Show MeSH
Related in: MedlinePlus