The Charles T. Campbell Eye Microbiology Lab
UPMCUniversity of Pittsburgh Schools of the Health Sciences
HomeContact InformationLab Diagnostic TestingAntibiotic SusceptibilityAntimicrobial TherapyCurrent ResearchPhotos


Ocular Microbiology and Immunology Group
Back to OMIG Main Page

< Previous | 2019 Agenda and Abstracts | Next >

 

2019 OMIG Abstract

IL-23 Drives Effector Th17/1 to Memory Th17 Cells in Dry Eye Disease

Nai-Wen Fan, Yihe Chen, Afsaneh Amouzegar, Reza Dana
Schepens Eye Research Institute, Massachusetts Eye and Ear, Department of Ophthalmology,
Harvard Medical School, Boston, MA

Purpose: Dry eye disease (DED) is one of the most prevalent ocular surface diseases. Immune-mediated inflammation plays a critical role in disease induction and chronicity. Our previous work has identified two subsets of effector T helper 17 that drive acute DED: effector Th17/1 (eTh17/1; CD4+IL-17+IFN-Y+) and effector Th17 (eTh17; CD4+IL-17+IFN-Y-); in contrast, memory Th17 cells (mTh17) perpetuate the chronic inflammation. However, it remains unknown which effector subset(s) serve as the precursor cells for memory Th17. In this study, we investigated the roles of IL-23 in driving eTh17/1 and eTh17 subsets into memory pool in DED.

Methods: Na´ve CD4+ cells were isolated from wild-type (WT) and T-bet null (no IFN-Y expression) mice, and then adoptively transferred into Rag1 -/- mice, respectively. The Rag1 -/- recipients were subsequently exposed to desiccating stress for 14 days followed by housed in the standard vivarium for additional 3 weeks. At the end of the follow up, the bulbar conjunctiva and DLNs of Rag1 -/- mice were collected, and mTh17 frequencies were examined by flow cytometry. Furthermore, eTh17 and eTh17/1 cells were isolated from DLNs of WT DED mice and cultured in medium supplemented with IL-23 for 48 hours, and the frequencies of mTh17 cells were assessed by flow cytometry. To evaluate the in vivo blockade effect, IL-23 signaling was blocked by intraperitoneal injection of anti-IL-23R antibody at day 14 when acute DED mice were returned to the normal environment. The disease severity was then evaluated for 7 days by corneal fluorescein staining, and mTh17 cells were examined using flow cytometry.

Results: Rag1 -/- mice receiving na´ve CD4+ cells isolated from T-bet null mice did not generate eTh17/1 cells by day 14, and developed significantly lower frequencies of mTh17 in both DLNs and conjunctiva by day 35 (chronic DED), compared to those receiving WT na´ve CD4+ cells (p<0.05). In vitro culture of eTh17/1 in the presence of IL-23 led to a prominent generation of mTh17 as 2 times high as eTh17 culture. Finally, in vivo blockade of IL-23 signaling prevented the development of mTh17 cells from existing effector Th17 cells, and significantly decreased ongoing disease severity by 33% (p = 0.004).

Conclusion: Our data suggest that eTh17/1 is the predominant effector subset converting to mTh17 mediated by IL-23, and blockade of IL-23 could be a promising therapeutic strategy for chronic DED.

Disclosure: N. This study was supported by grant: NIH R01 EY20889.

 

 

<Previous | 2019 Agenda and Abstracts | Next >

 


 

 

 

space