Effects of rat mesenchymal stem cell-derived exosomes on rat experimental autoimmune uveitis_Chinese Journal of Ocular Fundus Diseases

Authors：

Xie Ruotian , Bai Lingling , Yang Jing , Li Yongtao , Dong Lijie , Ma Feifei , Li Xiaorong ,  Zhang Xiaomin

Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, Tianjin Medical University School of Optometry and Ophthalmology, Tianjin 300384, China (Bai Lingling, now working at Department of Ophtalmology, Beijing ChuiYangLiu Hospital Affiliated to Tsinghua University);

Corresponding author：

Zhang Xiaomin, Email: xiaomzh@126.com

Keywords：

Mesenchymal stem cells; Exosomes; Uveitis/therapy; Animal experimentation

DOI：

10.3760/cma.j.issn.1005-1015.2018.06.008

Video：

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ObjectiveTo observe the effects of exosomes derived from rat mesenchymal stem cells (MSC-exosomes) on the rat experimental autoimmune uveitis (EAU) model.MethodsTwelve Lewis rats were randomly divided into experimental group and control group by random number table, with 6 rats in each group. Rats in the experimental group were established with EAU model, 100 μl of MSC-exosomes (50 μg) were periocular injected on the 9th day after modeling while the control rats were injected with the same volume of phosphate buffer. At different time points after modeling, the retinal structure was observed by hematoxylin and eosin (HE) staining, and the clinical and pathological manifestations were evaluated. T cells from the two groups were analyzed by flow cytometry. Immunohistochemical staining was used to observe the expression of macrophage surface marker CD68. The effect of MSC-exosomes on T cells was measured by lymphocyte proliferation assays. And flow cytometry was used to detect Th1, Th17 and regulatory T cells Variety. Electroretinogram (ERG) was used to evaluate the retinal function. Data were compared between the two groups using the t test.ResultsHE staining showed that the retina structure of the experimental group was more complete than that of the control group on the 15th day after modeling. Immunohistochemical staining showed that the positive expression of CD68 in the experimental group was significantly less than that in the control group. On the 15th day after modeling, the retinal pathological score of the experimental group was lower than that of the control group. On the 9th to 13th day after modeling, compared to the control group, the average clinical scores of the retina in the experimental group were lower, and the difference was statistically significant (t=3.665, 3.21, 3.181, 4.121, 3.227; P＜0.01). The results of T cell proliferation assay showed that exosomes (1.0, 10.0 μg/ml) inhibited the proliferation of T cells under different concentrations of R16 (1, 10, 30 μg/ml), and the difference was statistically significant (F=11.630, 4.188, 6.011; P＜0.05). The results of flow cytometry showed that the number of Th1, Th17 and Treg cell subsets in the experimental group was decreased compared with the control group, and the difference was statistically significant (t=7.374, 4.525, 6.910; P＜0.01). There was no difference in the proportion of cells in the T cells and lymph nodes (t=1.126, 0.493, 0.178; P=0.286, 0.632, 0.862). The results of ERG showed that, compared with the control group, the amplitudes of 0.01, 3.0 cd/m² a wave and b wave of the experiment group were all increased on the 15th day after modeling, and the differences were statistically significant (t=3.604, 4.178, 4.551, 2.566, P＜0.05).ConclusionsMSC-exosomes can reduce the clinical and pathological manifestations of EAU, protect retinal function, reduce ocular macrophage infiltration, down-regulate the proportion of inflammatory cells in the eye, and inhibit T cell proliferation.

Citation： Xie Ruotian, Bai Lingling, Yang Jing, Li Yongtao, Dong Lijie, Ma Feifei, Li Xiaorong, Zhang Xiaomin. Effects of rat mesenchymal stem cell-derived exosomes on rat experimental autoimmune uveitis. Chinese Journal of Ocular Fundus Diseases, 2018, 34(6): 562-567. doi: 10.3760/cma.j.issn.1005-1015.2018.06.008 Copy

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11.	Caspi RR. Experimental autoimmune uveoretinitis in the rat and mouse[J]. Curr Protoc Immunol, 2003, Chapter 15: Unit 15, 16. DOI: 10.1002/0471142735.im1506s53.
12.	Li G, Yuan L, Ren X, et al. The effect of mesenchymal stem cells on dynamic changes of T cell subsets in experimental autoimmune uveoretinitis[J]. Clin Exp Immunol, 2013, 173(1): 28-37. DOI: 10.1111/cei.12080.
13.	Treacy O, O'Flynn L, Ryan AE, et al. Mesenchymal stem cell therapy promotes corneal allograft survival in rats by local and systemic immunomodulation[J]. Am J Transplant, 2014, 14(9): 2023-2036. DOI: 10.1111/ajt.12828.
14.	Bai L, Shao H, Wang H, et al. Effects of mesenchymal stem cell-derived exosomes on experimental autoimmune uveitis[J]. Sci Rep, 2017, 7(1): 4323. DOI: 10.1038/s41598-017-04559-y.
15.	Xin H, Li Y, Liu Z, et al. MiR-133b promotes neural plasticity and functional recovery after treatment of stroke with multipotent mesenchymal stromal cells in rats via transfer of exosome-enriched extracellular particles[J]. Stem Cells, 2013, 31(12): 2737-2746. DOI: 10.1002/stem.1409.
16.	Ju GQ, Cheng J, Zhong L, et al. Microvesicles derived from human umbilical cord mesenchymal stem cells facilitate tubular epithelial cell dedifferentiation and growth via hepatocyte growth factor induction[J/OL]. PLoS One, 2015, 10(3): 0121534[2015-05-20]. http://dx.plos.org/10.1371/journal.pone.0121534. DOI: 10.1371/journal.pone.0121534.
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18.	Del Fattore A, Luciano R, Pascucci L, et al. Immunoregulatory effects of mesenchymal stem cell-derived extracellular vesicles on T lymphocytes[J]. Cell Transplant, 2015, 24(12): 2615-2627. DOI: 10.3727/096368915X687543.

1. Tempest-Roe S, Joshi L, Dick AD, et al. Local therapies for inflammatory eye disease in translation: past, present and future[J]. BMC Ophthalmol, 2013, 13(1): 39. DOI: 10.1186/1471-2415-13-39.
2. Rosenbaum JT. Future for biological therapy for uveitis[J]. Curr Opin Ophthalmol, 2010, 21(6): 473-477. DOI: 10.1097/ICU.0b013e32833f00b3.
3. Gonzalez MA, Gonzalez-Rey E, Rico L, et al. Treatment of experimental arthritis by inducing immune tolerance with human adipose-derived mesenchymal stem cells[J]. Arthritis Rheum, 2009, 60(4): 1006-1019. DOI: 10.1002/art.24405.
4. Zhao S, Wehner R, Bornhauser M, et al. Immunomodulatory properties of mesenchymal stromal cells and their therapeutic consequences for immune-mediated disorders[J]. Stem Cells Dev, 2010, 19(5): 607-614. DOI: 10.1089/scd.2009.0345.
5. Zhang J, Brodie C, Li Y, et al. Bone marrow stromal cell therapy reduces proNGF and p75 expression in mice with experimental autoimmune encephalomyelitis[J]. J Neurol Sci, 2009, 279(1-2): 30-38. DOI: 10.1016/j.jns.2008.12.033.
6. Vercelli A, Mereuta OM, Garbossa D, et al. Human mesenchymal stem cell transplantation extends survival, improves motor performance and decreases neuroinflammation in mouse model of amyotrophic lateral sclerosis[J]. Neurobiol Dis, 2008, 31(3): 395-405. DOI: 10.1016/j.nbd.2008.05.016.
7. Chen X, Shao H, Zhi Y, et al. CD73 pathway contributes to the immunosuppressive ability of mesenchymal stem cells in intraocular autoimmune responses[J]. Stem Cells Dev, 2016, 25(4): 337-346. DOI: 10.1089/scd.2015.0227.
8. Zhang Y, Zhang M, Zhao S, et al. Effects of human umbilical cord-derived mesenchymal stem cells on anterior chamber-associated immune deviation[J]. Int Immunopharmacol, 2013, 15(1): 114-120. DOI: 10.1016/j.intimp.2012.11.007.
9. Jia Z, Jiao C, Zhao S, et al. Immunomodulatory effects of mesenchymal stem cells in a rat corneal allograft rejection model[J]. Exp Eye Res, 2012, 102: 44-49. DOI: 10.1016/j.exer.2012.06.008.
10. Amarnath S, Foley JE, Farthing DE, et al. Bone marrow-derived mesenchymal stromal cells harness purinergenic signaling to tolerize human Th1 cells in vivo[J]. Stem Cells, 2015, 33(4): 1200-1212. DOI: 10.1002/stem.1934.
11. Caspi RR. Experimental autoimmune uveoretinitis in the rat and mouse[J]. Curr Protoc Immunol, 2003, Chapter 15: Unit 15, 16. DOI: 10.1002/0471142735.im1506s53.
12. Li G, Yuan L, Ren X, et al. The effect of mesenchymal stem cells on dynamic changes of T cell subsets in experimental autoimmune uveoretinitis[J]. Clin Exp Immunol, 2013, 173(1): 28-37. DOI: 10.1111/cei.12080.
13. Treacy O, O'Flynn L, Ryan AE, et al. Mesenchymal stem cell therapy promotes corneal allograft survival in rats by local and systemic immunomodulation[J]. Am J Transplant, 2014, 14(9): 2023-2036. DOI: 10.1111/ajt.12828.
14. Bai L, Shao H, Wang H, et al. Effects of mesenchymal stem cell-derived exosomes on experimental autoimmune uveitis[J]. Sci Rep, 2017, 7(1): 4323. DOI: 10.1038/s41598-017-04559-y.
15. Xin H, Li Y, Liu Z, et al. MiR-133b promotes neural plasticity and functional recovery after treatment of stroke with multipotent mesenchymal stromal cells in rats via transfer of exosome-enriched extracellular particles[J]. Stem Cells, 2013, 31(12): 2737-2746. DOI: 10.1002/stem.1409.
16. Ju GQ, Cheng J, Zhong L, et al. Microvesicles derived from human umbilical cord mesenchymal stem cells facilitate tubular epithelial cell dedifferentiation and growth via hepatocyte growth factor induction[J/OL]. PLoS One, 2015, 10(3): 0121534[2015-05-20]. http://dx.plos.org/10.1371/journal.pone.0121534. DOI: 10.1371/journal.pone.0121534.
17. Blazquez R, Sanchez-Margallo FM, de la Rosa O, et al. Immunomodulatory potential of human adipose mesenchymal stem cells derived exosomes on in vitro stimulated T cells[J]. Front Immunol, 2014, 5: 556. DOI: 10.3389/fimmu.2014.00556.
18. Del Fattore A, Luciano R, Pascucci L, et al. Immunoregulatory effects of mesenchymal stem cell-derived extracellular vesicles on T lymphocytes[J]. Cell Transplant, 2015, 24(12): 2615-2627. DOI: 10.3727/096368915X687543.

Chinese Journal of Ocular Fundus Diseases

Effects of rat mesenchymal stem cell-derived exosomes on rat experimental autoimmune uveitis

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