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Renyan Wang, ; Establishment and Regulation of the B lymphocytes and Orbital Fibroblasts Co-culture in Thyroid Associated Ophthalmopathy. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1843.
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© ARVO (1962-2015); The Authors (2016-present)
1.To establish and to identify an in vitro experimental model of autoimmune TAO by co-culture the orbital fibroblasts and peripheral B lymphocytes. 2.To investigate the Insulin-like growth factor-1(IGF-1R) receptor expressing on OFs, observe the inflammatory responses in the model, and explore the important role of B cells in TAO. 3.Exploring pathogenesis and therapeutic approaches for TAO.
1.Orbital fibroblasts obtained from 15 patients with TAO and from 15 control subjects were used to set up primary cultures, and were identified by immunohistochemistry. B lymphocytes were isolated from peripheral blood obtained from 10 patients with TAO and 10 controls. B cells were enriched, purified using immunomagnetic beads separation techniques and analyzed by folw cytometry. 2.The IGF-1R of OFs were evaluated by flow cytometry and observed under confocal microscopy. The expression of interleukin-6 and chemoattractants RANTES of were quantified by ELISA at 24hours,48 hours and 72 hours after co-culture. 3.The depleting effects of Rituximab on B cells at several concentrations in different time were measured by MTS. The inhibition on the expression of IL-6 and RANTES after using RTX and IGF-1 binding protein on the co-culture model were analyzed by ELISA.
1.The co-culture of orbital fibroblasts and B lymphocytes was established. 2.The expression of IGF-1R on OFs in patients with TAO was significantly higher than normal’s. 3.The expression of IL-6 and RANTES in each co-culture group was increased at 24 hours, especially in the T+T group (B lymphocytes and orbital fibroblasts all obtained from the TAO patients) is the highest. 4.RTX and IGF-1 binding protein significantly inhibited the expression of IL-6 and RANTES in the T+T co-culture group at 48 hours.
1.An in vitro model that partially represents TAO, as an autoimmune inflammatory disease, was established by co-culture of orbital fibroblasts and peripheral B lymphoctes. The interactions between the two cells may play a role in TAO pathogenesis. 2.IGF-1R may be the pathway in the interactions between OFs and B cells. It may mediate the process of TAO. Inflammatory responses may be depressed after blocking the recptor. 3.B cell depleting agents RTX have a good anti-inflammatory effects in the vitro model. Monoclonal anti-CD20 antibody therapy may be a novel treatment option in TAO in the future.
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