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Ex-vivo tolerogenic F4/80⁺ antigen-presenting cells (APC) induce efferent CD8⁺ regulatory T cell-dependent suppression of experimental autoimmune uveitis.

Hsu SM, Mathew R, Taylor AW, Stein-Streilein J - Clin. Exp. Immunol. (2014)

Bottom Line: We observed that retinal antigen-pulsed TolAPC suppressed the incidence and severity of the clinical expression of EAU and reduced the expression of associated inflammatory cytokines.Retinal antigen-pulsed TolAPC suppressed ongoing EAU by inducing CD8⁺ T(reg) cells that, in turn, suppressed the effector activity of the IRBP-specific T cells and altered the clinical symptoms of autoimmune inflammation in the eye.The ability to use retinal extract for the antigen raises the possibility that retinal extract could be used to produce autologous TolAPC and then used as therapy in human uveitis.

View Article: PubMed Central - PubMed

Affiliation: Schepens Eye Research Institute, Department of Ophthalmology, Harvard Medical School, Boston, MA, USA; Department of Ophthalmology, National Cheng-Kung University Hospital, Tainan City, Taiwan.

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Effect of CD8+ tumour necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)– cells on suppression. (a) Flow cytometry gate used for analysis. Spleen cells were cultured with antigen-presenting cells (APC) treated with transforming growth factor (TGF)-β2 and interphotoreceptor retinoid-binding protein (IRBP). Seven days later the CD8+ T cells were enriched by magnetic beads and single-stained for CD8 (top panel). Lower panel: an aliquot of enriched CD8+ T cells was stained with TRAIL phycoerythrin (PE) (abscissa) and forkhead box protein 3 (FoxP3) (fluorescein isothiocyanate (FITC) (ordinate). (b) Local adoptive transfer assay using CD8+TRAIL– T cells. The experiment was performed twice with similar results. *Significant difference (P ≤ 0·05).
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fig05: Effect of CD8+ tumour necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)– cells on suppression. (a) Flow cytometry gate used for analysis. Spleen cells were cultured with antigen-presenting cells (APC) treated with transforming growth factor (TGF)-β2 and interphotoreceptor retinoid-binding protein (IRBP). Seven days later the CD8+ T cells were enriched by magnetic beads and single-stained for CD8 (top panel). Lower panel: an aliquot of enriched CD8+ T cells was stained with TRAIL phycoerythrin (PE) (abscissa) and forkhead box protein 3 (FoxP3) (fluorescein isothiocyanate (FITC) (ordinate). (b) Local adoptive transfer assay using CD8+TRAIL– T cells. The experiment was performed twice with similar results. *Significant difference (P ≤ 0·05).

Mentions: After co-culturing F4/80+ TolAPC with spleen cells for 7 days, the non-adherent cells were harvested and the CD8+ T cells sorted into TRAIL-negative and -positive populations (Fig. 5a). The CD8+TRAIL– T cells suppressed the response to IRBP in a LAT assay (Fig. 5b) (there were insufficient CD8+TRAIL+ cells to test their function). Flow analysis of the TRAIL–CD8+ Treg cells confirmed that CD8+ Treg cells expressed CD103 (data not shown) 22. Thus, 7-day cultures of TolAPC with spleen cells generate CD8+CD103+FoxP3+TRAIL– Treg cells that are capable of suppressing efferent immune responses in vivo.


Ex-vivo tolerogenic F4/80⁺ antigen-presenting cells (APC) induce efferent CD8⁺ regulatory T cell-dependent suppression of experimental autoimmune uveitis.

Hsu SM, Mathew R, Taylor AW, Stein-Streilein J - Clin. Exp. Immunol. (2014)

Effect of CD8+ tumour necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)– cells on suppression. (a) Flow cytometry gate used for analysis. Spleen cells were cultured with antigen-presenting cells (APC) treated with transforming growth factor (TGF)-β2 and interphotoreceptor retinoid-binding protein (IRBP). Seven days later the CD8+ T cells were enriched by magnetic beads and single-stained for CD8 (top panel). Lower panel: an aliquot of enriched CD8+ T cells was stained with TRAIL phycoerythrin (PE) (abscissa) and forkhead box protein 3 (FoxP3) (fluorescein isothiocyanate (FITC) (ordinate). (b) Local adoptive transfer assay using CD8+TRAIL– T cells. The experiment was performed twice with similar results. *Significant difference (P ≤ 0·05).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3958152&req=5

fig05: Effect of CD8+ tumour necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)– cells on suppression. (a) Flow cytometry gate used for analysis. Spleen cells were cultured with antigen-presenting cells (APC) treated with transforming growth factor (TGF)-β2 and interphotoreceptor retinoid-binding protein (IRBP). Seven days later the CD8+ T cells were enriched by magnetic beads and single-stained for CD8 (top panel). Lower panel: an aliquot of enriched CD8+ T cells was stained with TRAIL phycoerythrin (PE) (abscissa) and forkhead box protein 3 (FoxP3) (fluorescein isothiocyanate (FITC) (ordinate). (b) Local adoptive transfer assay using CD8+TRAIL– T cells. The experiment was performed twice with similar results. *Significant difference (P ≤ 0·05).
Mentions: After co-culturing F4/80+ TolAPC with spleen cells for 7 days, the non-adherent cells were harvested and the CD8+ T cells sorted into TRAIL-negative and -positive populations (Fig. 5a). The CD8+TRAIL– T cells suppressed the response to IRBP in a LAT assay (Fig. 5b) (there were insufficient CD8+TRAIL+ cells to test their function). Flow analysis of the TRAIL–CD8+ Treg cells confirmed that CD8+ Treg cells expressed CD103 (data not shown) 22. Thus, 7-day cultures of TolAPC with spleen cells generate CD8+CD103+FoxP3+TRAIL– Treg cells that are capable of suppressing efferent immune responses in vivo.

Bottom Line: We observed that retinal antigen-pulsed TolAPC suppressed the incidence and severity of the clinical expression of EAU and reduced the expression of associated inflammatory cytokines.Retinal antigen-pulsed TolAPC suppressed ongoing EAU by inducing CD8⁺ T(reg) cells that, in turn, suppressed the effector activity of the IRBP-specific T cells and altered the clinical symptoms of autoimmune inflammation in the eye.The ability to use retinal extract for the antigen raises the possibility that retinal extract could be used to produce autologous TolAPC and then used as therapy in human uveitis.

View Article: PubMed Central - PubMed

Affiliation: Schepens Eye Research Institute, Department of Ophthalmology, Harvard Medical School, Boston, MA, USA; Department of Ophthalmology, National Cheng-Kung University Hospital, Tainan City, Taiwan.

Show MeSH
Related in: MedlinePlus