Tags

Type your tag names separated by a space and hit enter

Immunosuppressive properties of regulatory T cells generated by incubation of peripheral blood mononuclear cells with supernatants of human RPE cells.
Invest Ophthalmol Vis Sci. 2012 Oct 23; 53(11):7299-309.IO

Abstract

PURPOSE

To determine whether supernatants of human retinal pigment epithelium (RPE) cells can convert CD4⁺ T cells into regulatory T cells (Tregs) under Treg-induction conditions in vitro and in vivo.

METHODS

Peripheral blood mononuclear cells were cocultured with supernatants from TGFβ2-pretreated human RPE lines on anti-CD3-coated plates. Cells were then separated with a CD4⁺CD25⁺ Treg isolation kit and cultured with supernatants from RPE, anti-CD3/CD28 antibodies, high-dose IL-2, and TGFβ2. By flow cytometry sorting, CD25⁺CD45RA⁻ Tregs were separated. Expressions of CD25(high), Foxp3, CD152, and TNFRSF 18 on Tregs were analyzed by flow cytometry. Cytokine production by Tregs was measured by ELISA. Proliferation of target T cells was assessed by [³H]thymidine incorporation or CFSE incorporation. In addition, mouse RPE-induced Tregs were used for the in vitro assay and in vivo experimental autoimmune uveitis (EAU) models.

RESULTS

Human RPE-induced Tregs expressed higher levels of the Treg markers CD25(high), Foxp3, CD152, and TNFRSF 18. In addition, RPE-induced Tregs included significant numbers of CD4⁺CD25(high)Foxp3(high)CD45RA⁻ active effector Tregs that significantly suppressed the activation of Th1/Th17 cell lines, indicating that they have immunosuppressive properties. Furthermore, CD4⁺CD25(low)Foxp3(low)CD45RA⁻ nonsuppressing cytokine-secreting T cells were removed from the in vitro-manipulated Treg population. Administration of mouse RPE-induced Tregs significantly suppressed ocular inflammation in mice with EAU. In addition, the Tregs suppressed retinal antigen-specific T cells in vitro.

CONCLUSIONS

It is hoped that through the data provided in this study that Tregs might become useful as individualized therapeutic agents for ocular autoimmune diseases.

Authors+Show Affiliations

Department of Ophthalmology and Visual Science, Tokyo Medical and Dental University Graduate School of Medicine and Dental Sciences, Tokyo, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23036994

Citation

Imai, Ayano, et al. "Immunosuppressive Properties of Regulatory T Cells Generated By Incubation of Peripheral Blood Mononuclear Cells With Supernatants of Human RPE Cells." Investigative Ophthalmology & Visual Science, vol. 53, no. 11, 2012, pp. 7299-309.
Imai A, Sugita S, Kawazoe Y, et al. Immunosuppressive properties of regulatory T cells generated by incubation of peripheral blood mononuclear cells with supernatants of human RPE cells. Invest Ophthalmol Vis Sci. 2012;53(11):7299-309.
Imai, A., Sugita, S., Kawazoe, Y., Horie, S., Yamada, Y., Keino, H., Maruyama, K., & Mochizuki, M. (2012). Immunosuppressive properties of regulatory T cells generated by incubation of peripheral blood mononuclear cells with supernatants of human RPE cells. Investigative Ophthalmology & Visual Science, 53(11), 7299-309. https://doi.org/10.1167/iovs.12-10182
Imai A, et al. Immunosuppressive Properties of Regulatory T Cells Generated By Incubation of Peripheral Blood Mononuclear Cells With Supernatants of Human RPE Cells. Invest Ophthalmol Vis Sci. 2012 Oct 23;53(11):7299-309. PubMed PMID: 23036994.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Immunosuppressive properties of regulatory T cells generated by incubation of peripheral blood mononuclear cells with supernatants of human RPE cells. AU - Imai,Ayano, AU - Sugita,Sunao, AU - Kawazoe,Yuko, AU - Horie,Shintaro, AU - Yamada,Yukiko, AU - Keino,Hiroshi, AU - Maruyama,Kazuichi, AU - Mochizuki,Manabu, Y1 - 2012/10/23/ PY - 2012/10/6/entrez PY - 2012/10/6/pubmed PY - 2013/1/9/medline SP - 7299 EP - 309 JF - Investigative ophthalmology & visual science JO - Invest Ophthalmol Vis Sci VL - 53 IS - 11 N2 - PURPOSE: To determine whether supernatants of human retinal pigment epithelium (RPE) cells can convert CD4⁺ T cells into regulatory T cells (Tregs) under Treg-induction conditions in vitro and in vivo. METHODS: Peripheral blood mononuclear cells were cocultured with supernatants from TGFβ2-pretreated human RPE lines on anti-CD3-coated plates. Cells were then separated with a CD4⁺CD25⁺ Treg isolation kit and cultured with supernatants from RPE, anti-CD3/CD28 antibodies, high-dose IL-2, and TGFβ2. By flow cytometry sorting, CD25⁺CD45RA⁻ Tregs were separated. Expressions of CD25(high), Foxp3, CD152, and TNFRSF 18 on Tregs were analyzed by flow cytometry. Cytokine production by Tregs was measured by ELISA. Proliferation of target T cells was assessed by [³H]thymidine incorporation or CFSE incorporation. In addition, mouse RPE-induced Tregs were used for the in vitro assay and in vivo experimental autoimmune uveitis (EAU) models. RESULTS: Human RPE-induced Tregs expressed higher levels of the Treg markers CD25(high), Foxp3, CD152, and TNFRSF 18. In addition, RPE-induced Tregs included significant numbers of CD4⁺CD25(high)Foxp3(high)CD45RA⁻ active effector Tregs that significantly suppressed the activation of Th1/Th17 cell lines, indicating that they have immunosuppressive properties. Furthermore, CD4⁺CD25(low)Foxp3(low)CD45RA⁻ nonsuppressing cytokine-secreting T cells were removed from the in vitro-manipulated Treg population. Administration of mouse RPE-induced Tregs significantly suppressed ocular inflammation in mice with EAU. In addition, the Tregs suppressed retinal antigen-specific T cells in vitro. CONCLUSIONS: It is hoped that through the data provided in this study that Tregs might become useful as individualized therapeutic agents for ocular autoimmune diseases. SN - 1552-5783 UR - https://www.unboundmedicine.com/medline/citation/23036994/Immunosuppressive_properties_of_regulatory_T_cells_generated_by_incubation_of_peripheral_blood_mononuclear_cells_with_supernatants_of_human_RPE_cells_ L2 - https://iovs.arvojournals.org/article.aspx?doi=10.1167/iovs.12-10182 DB - PRIME DP - Unbound Medicine ER -