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Sunao Sugita, Shintaro Horie, Yukiko Yamada, Manabu Mochizuki; Inhibition of B-Cell Activation by Retinal Pigment Epithelium. Invest. Ophthalmol. Vis. Sci. 2010;51(11):5783-5788. doi: 10.1167/iovs.09-5098.
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To determine whether retinal pigment epithelial (RPE) cells can inhibit B-cell activation in vitro.
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 [3H]-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.
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.
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.
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