June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Proinflammatory Th17 cells are more effective than Th1 cells in providing B cell help in dry eye disease.
Author Affiliations & Notes
  • Brinda Subbarayal
    Opthalmology, Schepens Eye Research Institute, Quincy, MA
  • Antonio Di Zazzo
    Opthalmology, Schepens Eye Research Institute, Quincy, MA
  • Susanne Eiglmeier
    Opthalmology, Schepens Eye Research Institute, Quincy, MA
  • SANGMOK LEE
    Opthalmology, Schepens Eye Research Institute, Quincy, MA
  • Sunil Chauhan
    Opthalmology, Schepens Eye Research Institute, Quincy, MA
  • Reza Dana
    Opthalmology, Schepens Eye Research Institute, Quincy, MA
  • Footnotes
    Commercial Relationships Brinda Subbarayal, None; Antonio Di Zazzo, None; Susanne Eiglmeier, None; SANGMOK LEE, None; Sunil Chauhan, None; Reza Dana, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 4877. doi:
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      Brinda Subbarayal, Antonio Di Zazzo, Susanne Eiglmeier, SANGMOK LEE, Sunil Chauhan, Reza Dana; Proinflammatory Th17 cells are more effective than Th1 cells in providing B cell help in dry eye disease.. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):4877.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: T cell help is known to be fundamental for the induction and subsequent organization of germinal centers (GCs), resulting in the generation of memory B cells and long-lived plasma cells. However, it is not known whether Th17 cells can act as effective helper T cells in experimental dry eye disease (DED). The aim of this study was to determine whether Th17 cells function as helper cells for B cells in DED.

Methods: Dry eye was induced by exposing female C57BL/6 mice to desiccating stress (subcutaneous injection of scopolamine [0.5 mg/ 0.1 mL] 3 times a day, humidity < 15%, and airflow rate of 15L/min) for 3 weeks. To determine the kinetics of GC formation and isotype switched B cells, cervical lymph nodes were stained for GCs (B220+GL7+FAS+) and isotype switched (B220+IgD-IgM-) B cells on day 0, 7, 14, 21, and 28 using flow cytometry. On day 21, formation of GCs was analyzed by immunhistochemistcal staining of lymph node sections with anti-ki67-FITC and anti-IgM-APC antibodies. T cell-B cell coculture assays were performed to determine T cell help on B cells. The cells were activated with 1μg/ml of anti-CD3 and 1μg/ml of anti-IgM for 36hours before analyzing B cell proliferation using flow cytometry.

Results: Germinal center B cell formation in the lymph nodes of DED mice peaked on day 21, whereas no GC formation was observed in naïve mice. T effector (Teff) cells from DED mice significantly enhanced B cell proliferation in a contact dependent manner. To determine whether Th1<br /> (IFN-γ+) or Th17 (IL-17+) cells provide major B cell help, IFN-γ or IL-17 depleted Teff (CD4+CD25-) cells were cocultured with B cells. Although both subsets effectively reduced B cell proliferation in vitro, Th17 cells were more effective than Th1 cells in helping B cell proliferation in DED mice.

Conclusions: Th17 cells are more effective than Th1 cells in inducing B cell proliferation in DED.

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