April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Establishment and Regulation of the B lymphocytes and Orbital Fibroblasts Co-culture in Thyroid Associated Ophthalmopathy
Author Affiliations & Notes
  • Renyan Wang
    Ophthalmology, Beijing Tsinghua Chang Gung hospital, Beijing, China
    Ophthalmology, Peking Union Medical College Hospital, Beijing, China
  • Footnotes
    Commercial Relationships Renyan Wang, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 1843. doi:
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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)

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Abstract

Purpose: 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.

Methods: 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.

Results: 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.

Conclusions: 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.

Keywords: 637 pathology: experimental • 432 autoimmune disease • 554 immunohistochemistry  
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