Sunao Sugita1, Shintaro Horie, Yukiko Yamada, Manabu Mochizuki. 1. Department of Ophthalmology and Visual Science, Tokyo Medical and Dental University Graduate School of Medicine and Dental Sciences, Tokyo, Japan. sunaoph@tmd.ac.jp
Abstract
PURPOSE: To determine whether retinal pigment epithelial (RPE) cells can inhibit B-cell activation in vitro. METHODS: Primary cultured RPE cells were established from normal C57BL/6 mice. Activated target B cells were established from splenic B cells stimulated with anti-mouse CD40 antibody and lipopolysaccharide (LPS) in the presence of recombinant interleukin 4 (rIL-4). B-cell activation was assessed by examining proliferation through [(3)H]-thymidine incorporation or carboxyfluorescein succinimidyl ester dilution, and antibody production was determined by ELISA. Expression of costimulatory molecules and the receptors on B cells was evaluated by flow cytometry. Neutralizing anti-TGFβ antibodies were used in the assay. RESULTS: Addition of primary cultured RPE cells suppressed B-cell proliferation in response to anti-CD40, LPS, and rIL-4 stimulation. Similarly, antibody production by these activated B cells was suppressed. Suppression of B-cell activation was mediated by a soluble factor because supernatants from cultured RPE cells were sufficient to inhibit B-cell responses. Moreover, TGFβ was identified as the soluble mediator given that RPE-supernatants failed to suppress B-cell activation if pretreated with neutralizing anti-TGFβ antibodies. CONCLUSIONS: Cultured RPE cells suppress the activation of B cells in vitro. These data support the hypothesis that retinal pigment epithelium has immunosuppressive properties that are capable of suppressing B-cell activation.
PURPOSE: To determine whether retinal pigment epithelial (RPE) cells can inhibit B-cell activation in vitro. METHODS: Primary cultured RPE cells were established from normal C57BL/6 mice. Activated target B cells were established from splenic B cells stimulated with anti-mouseCD40 antibody and lipopolysaccharide (LPS) in the presence of recombinant interleukin 4 (rIL-4). B-cell activation was assessed by examining proliferation through [(3)H]-thymidine incorporation or carboxyfluorescein succinimidyl ester dilution, and antibody production was determined by ELISA. Expression of costimulatory molecules and the receptors on B cells was evaluated by flow cytometry. Neutralizing anti-TGFβ antibodies were used in the assay. RESULTS: Addition of primary cultured RPE cells suppressed B-cell proliferation in response to anti-CD40, LPS, and rIL-4 stimulation. Similarly, antibody production by these activated B cells was suppressed. Suppression of B-cell activation was mediated by a soluble factor because supernatants from cultured RPE cells were sufficient to inhibit B-cell responses. Moreover, TGFβ was identified as the soluble mediator given that RPE-supernatants failed to suppress B-cell activation if pretreated with neutralizing anti-TGFβ antibodies. CONCLUSIONS: Cultured RPE cells suppress the activation of B cells in vitro. These data support the hypothesis that retinal pigment epithelium has immunosuppressive properties that are capable of suppressing B-cell activation.
Authors: Xiaoman Li; Xiaowu Gu; Timothy M Boyce; Min Zheng; Alaina M Reagan; Hui Qi; Nawajes Mandal; Alex W Cohen; Michelle C Callegan; Daniel J J Carr; Michael H Elliott Journal: Invest Ophthalmol Vis Sci Date: 2014-08-26 Impact factor: 4.799
Authors: Amanda L Piquet; Kala Venkiteswaran; Neena I Marupudi; Matthew Berk; Thyagarajan Subramanian Journal: Brain Res Bull Date: 2012-04-11 Impact factor: 4.077
Authors: Justine R Smith; Shawn Todd; Liam M Ashander; Theodosia Charitou; Yuefang Ma; Steven Yeh; Ian Crozier; Michael Z Michael; Binoy Appukuttan; Keryn A Williams; David J Lynn; Glenn A Marsh Journal: Transl Vis Sci Technol Date: 2017-07-14 Impact factor: 3.283