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Intraperitoneal infusion of mesenchymal stem/stromal cells prevents experimental autoimmune uveitis in mice.

Oh JY, Kim TW, Jeong HJ, Lee HJ, Ryu JS, Wee WR, Heo JW, Kim MK - Mediators Inflamm. (2014)

Bottom Line: The hMSCs did not reduce the levels of IL-1β, IL-6, IL-12, and IL-23 which are the cytokines that drive Th1/Th17 differentiation.Also, hMSCs did not induce CD4(+)CD25(+)Foxp3(+) cells.Our results support suggestions that hMSCs may offer a therapy for autoimmune diseases mediated by Th1/Th17 responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Seoul National University Hospital, Seoul 110-744, Republic of Korea ; Laboratory of Ocular Regenerative Medicine and Immunology, Seoul Artificial Eye Center, Seoul National University Hospital Biomedical Research Institute, Seoul 110-744, Republic of Korea.

ABSTRACT
Autoimmune uveitis is one of the leading causes of blindness. We here investigated whether intraperitoneal administration of human mesenchymal stem/stromal cells (hMSCs) might prevent development of experimental autoimmune uveitis (EAU) in mice. Time course study showed that the number of IFN-γ- or IL-17-expressing CD4(+) T cells was increased in draining lymph nodes (DLNs) on the postimmunization day 7 and decreased thereafter. The retinal structure was severely disrupted on day 21. An intraperitoneal injection of hMSCs at the time of immunization protected the retina from damage and suppressed the levels of proinflammatory cytokines in the eye. Analysis of DLNs on day 7 showed that hMSCs decreased the number of Th1 and Th17 cells. The hMSCs did not reduce the levels of IL-1β, IL-6, IL-12, and IL-23 which are the cytokines that drive Th1/Th17 differentiation. Also, hMSCs did not induce CD4(+)CD25(+)Foxp3(+) cells. However, hMSCs increased the level of an immunoregulatory cytokine IL-10 and the population of IL-10-expressing B220(+)CD19(+) cells. Together, data demonstrate that hMSCs attenuate EAU by suppressing Th1/Th17 cells and induce IL-10-expressing B220(+)CD19(+) cells. Our results support suggestions that hMSCs may offer a therapy for autoimmune diseases mediated by Th1/Th17 responses.

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Histological findings of the eye. (a) EAU was induced in mice on day 0, and either hMSCs (1 × 106 cells in 100 μL BSS) or BSS (100 μL) were intraperitoneally (IP) injected immediately after EAU induction. On days 7, 14, and 21, the eyes or DLNs were collected for assays. (b) ELISA showed that IFN-γ in the eye was markedly increased in the eyeball on day 14 and significantly reduced by hMSCs. Data are presented in mean + SEM. n = 5 in each group. (c) Time course of histological disease scores demonstrated that the retinal pathology gradually developed with a peak at day 21. Histological scores were significantly lower in hMSCs-treated mice at all time-points, suggesting that hMSCs prevented the disease development. Data are presented in mean + SEM. n = 5 in each group. (d) Hematoxylin-eosin staining of the eye on day 21 showed severe disruption of the retinal structure including the photoreceptor layer with inflammatory cell infiltration in the vitreous cavity and in the retina of EAU mice. In contrast, the retinal structure was well-reserved, and few inflammatory cells were observed in EAU mice treated with hMSCs. (e) TUENL staining showed a number of dead cells in the disrupted photoreceptor layer of EAU mice. In contrast, no TUENL-positive cells were found in the retina of mice treated with hMSCs.
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fig1: Histological findings of the eye. (a) EAU was induced in mice on day 0, and either hMSCs (1 × 106 cells in 100 μL BSS) or BSS (100 μL) were intraperitoneally (IP) injected immediately after EAU induction. On days 7, 14, and 21, the eyes or DLNs were collected for assays. (b) ELISA showed that IFN-γ in the eye was markedly increased in the eyeball on day 14 and significantly reduced by hMSCs. Data are presented in mean + SEM. n = 5 in each group. (c) Time course of histological disease scores demonstrated that the retinal pathology gradually developed with a peak at day 21. Histological scores were significantly lower in hMSCs-treated mice at all time-points, suggesting that hMSCs prevented the disease development. Data are presented in mean + SEM. n = 5 in each group. (d) Hematoxylin-eosin staining of the eye on day 21 showed severe disruption of the retinal structure including the photoreceptor layer with inflammatory cell infiltration in the vitreous cavity and in the retina of EAU mice. In contrast, the retinal structure was well-reserved, and few inflammatory cells were observed in EAU mice treated with hMSCs. (e) TUENL staining showed a number of dead cells in the disrupted photoreceptor layer of EAU mice. In contrast, no TUENL-positive cells were found in the retina of mice treated with hMSCs.

Mentions: EAU was induced in mice by subcutaneous injection of IRBP in a footpad on day 0. Simultaneously, either hMSCs (1 × 106 cells/mouse) or BSS was injected intraperitoneally. On days 7, 14, and 21, the mice were humanely killed, and eyeballs and draining LNs (DLNs) were collected for assays (Figure 1(a)). ELISA showed that the level of the proinflammatory cytokine IFN-γ was markedly increased in the eyeball on day 14 and significantly reduced by treatment with hMSCs (Figure 1(b)). Histology demonstrated that the retinal structure including the photoreceptor layer was severely disorganized with massive infiltration of inflammatory cells in the vitreous cavity and in the retina of BSS-treated EAU mice on day 21 (histological score 2.13 ± 0.31, Figures 1(c) and 1(d)). In contrast, the retinal architecture was almost completely preserved with few inflammatory cells in hMSCs-treated mice on day 21 (histological score 0.25 ± 0.14, Figures 1(c) and 1(d)). Similarly, TUNEL staining indicated the presence of many dead cells in the photoreceptor layer in BSS-treated EAU mice on day 21, while there were few TUNEL-positive cells in mice treated with hMSCs (Figure 1(e)).


Intraperitoneal infusion of mesenchymal stem/stromal cells prevents experimental autoimmune uveitis in mice.

Oh JY, Kim TW, Jeong HJ, Lee HJ, Ryu JS, Wee WR, Heo JW, Kim MK - Mediators Inflamm. (2014)

Histological findings of the eye. (a) EAU was induced in mice on day 0, and either hMSCs (1 × 106 cells in 100 μL BSS) or BSS (100 μL) were intraperitoneally (IP) injected immediately after EAU induction. On days 7, 14, and 21, the eyes or DLNs were collected for assays. (b) ELISA showed that IFN-γ in the eye was markedly increased in the eyeball on day 14 and significantly reduced by hMSCs. Data are presented in mean + SEM. n = 5 in each group. (c) Time course of histological disease scores demonstrated that the retinal pathology gradually developed with a peak at day 21. Histological scores were significantly lower in hMSCs-treated mice at all time-points, suggesting that hMSCs prevented the disease development. Data are presented in mean + SEM. n = 5 in each group. (d) Hematoxylin-eosin staining of the eye on day 21 showed severe disruption of the retinal structure including the photoreceptor layer with inflammatory cell infiltration in the vitreous cavity and in the retina of EAU mice. In contrast, the retinal structure was well-reserved, and few inflammatory cells were observed in EAU mice treated with hMSCs. (e) TUENL staining showed a number of dead cells in the disrupted photoreceptor layer of EAU mice. In contrast, no TUENL-positive cells were found in the retina of mice treated with hMSCs.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig1: Histological findings of the eye. (a) EAU was induced in mice on day 0, and either hMSCs (1 × 106 cells in 100 μL BSS) or BSS (100 μL) were intraperitoneally (IP) injected immediately after EAU induction. On days 7, 14, and 21, the eyes or DLNs were collected for assays. (b) ELISA showed that IFN-γ in the eye was markedly increased in the eyeball on day 14 and significantly reduced by hMSCs. Data are presented in mean + SEM. n = 5 in each group. (c) Time course of histological disease scores demonstrated that the retinal pathology gradually developed with a peak at day 21. Histological scores were significantly lower in hMSCs-treated mice at all time-points, suggesting that hMSCs prevented the disease development. Data are presented in mean + SEM. n = 5 in each group. (d) Hematoxylin-eosin staining of the eye on day 21 showed severe disruption of the retinal structure including the photoreceptor layer with inflammatory cell infiltration in the vitreous cavity and in the retina of EAU mice. In contrast, the retinal structure was well-reserved, and few inflammatory cells were observed in EAU mice treated with hMSCs. (e) TUENL staining showed a number of dead cells in the disrupted photoreceptor layer of EAU mice. In contrast, no TUENL-positive cells were found in the retina of mice treated with hMSCs.
Mentions: EAU was induced in mice by subcutaneous injection of IRBP in a footpad on day 0. Simultaneously, either hMSCs (1 × 106 cells/mouse) or BSS was injected intraperitoneally. On days 7, 14, and 21, the mice were humanely killed, and eyeballs and draining LNs (DLNs) were collected for assays (Figure 1(a)). ELISA showed that the level of the proinflammatory cytokine IFN-γ was markedly increased in the eyeball on day 14 and significantly reduced by treatment with hMSCs (Figure 1(b)). Histology demonstrated that the retinal structure including the photoreceptor layer was severely disorganized with massive infiltration of inflammatory cells in the vitreous cavity and in the retina of BSS-treated EAU mice on day 21 (histological score 2.13 ± 0.31, Figures 1(c) and 1(d)). In contrast, the retinal architecture was almost completely preserved with few inflammatory cells in hMSCs-treated mice on day 21 (histological score 0.25 ± 0.14, Figures 1(c) and 1(d)). Similarly, TUNEL staining indicated the presence of many dead cells in the photoreceptor layer in BSS-treated EAU mice on day 21, while there were few TUNEL-positive cells in mice treated with hMSCs (Figure 1(e)).

Bottom Line: The hMSCs did not reduce the levels of IL-1β, IL-6, IL-12, and IL-23 which are the cytokines that drive Th1/Th17 differentiation.Also, hMSCs did not induce CD4(+)CD25(+)Foxp3(+) cells.Our results support suggestions that hMSCs may offer a therapy for autoimmune diseases mediated by Th1/Th17 responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Seoul National University Hospital, Seoul 110-744, Republic of Korea ; Laboratory of Ocular Regenerative Medicine and Immunology, Seoul Artificial Eye Center, Seoul National University Hospital Biomedical Research Institute, Seoul 110-744, Republic of Korea.

ABSTRACT
Autoimmune uveitis is one of the leading causes of blindness. We here investigated whether intraperitoneal administration of human mesenchymal stem/stromal cells (hMSCs) might prevent development of experimental autoimmune uveitis (EAU) in mice. Time course study showed that the number of IFN-γ- or IL-17-expressing CD4(+) T cells was increased in draining lymph nodes (DLNs) on the postimmunization day 7 and decreased thereafter. The retinal structure was severely disrupted on day 21. An intraperitoneal injection of hMSCs at the time of immunization protected the retina from damage and suppressed the levels of proinflammatory cytokines in the eye. Analysis of DLNs on day 7 showed that hMSCs decreased the number of Th1 and Th17 cells. The hMSCs did not reduce the levels of IL-1β, IL-6, IL-12, and IL-23 which are the cytokines that drive Th1/Th17 differentiation. Also, hMSCs did not induce CD4(+)CD25(+)Foxp3(+) cells. However, hMSCs increased the level of an immunoregulatory cytokine IL-10 and the population of IL-10-expressing B220(+)CD19(+) cells. Together, data demonstrate that hMSCs attenuate EAU by suppressing Th1/Th17 cells and induce IL-10-expressing B220(+)CD19(+) cells. Our results support suggestions that hMSCs may offer a therapy for autoimmune diseases mediated by Th1/Th17 responses.

Show MeSH
Related in: MedlinePlus