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2014 Agenda and Abstracts | < Previous | Next >

2014 OMIG Abstract 23

Proinflammatory Th1 and Th17 Cells Provide B Cell Help in Dry Eye Disease
Brinda Subbarayal, Antonio Di Zazzo, Sang-Mok Lee, Sunil Chauhan, Reza Dana
Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, MA

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 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 GC (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 staining lymph node sections with ki67-FITC and IgM-APC using immunhistochemistry. To determine T cell help on B cells, T cell-B cell coculture assays were performed and cells were activated with 1μg/ml of anti-CD3 and 1μg/ml of anti-IgM for 36hrs before analyzing B cell proliferation using flow cytometry.

Results: GC B cell formation in the LNs of DED mice peaked on day 21, whereas no GC formation was observed in naïve mice. Teffector (Teff) cells from DED mice significantly enhanced B cells in a contact dependent manner. To determine which subset (Th1 or Th17) provides major B cell help, IFN-γ or IL-17 depleted Teff (CD4+CD25-) cells were cocultured with B cells, and both Th1 and Th17 depleted Teff cells effectively reduced B cell proliferation in vitro.

Conclusion: Both Th1 and Th17 cells act as effective helper cells for B cells in dry eye disease.

Disclosure: BS:N; AZ: N; SL:N; SC:N; RD:S
Grant Support: NIH Grant R01-EY-20889

2014 Agenda and Abstracts | < Previous | Next >