June 2020
Volume 61, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2020
Effect of Dry Eye Disease on the Kinetics of Lacrimal Gland Dendritic Cells as Visualized by Intravital Multi-Photon Microscopy
Author Affiliations & Notes
  • Gustavo Ortiz
    Center for Translational Ocular Immunology, Department of Ophthalmology, Tufts Medical Center, Tufts University School of Medicine, Boston, Massachusetts, United States
  • Cecilia Chao
    Center for Translational Ocular Immunology, Department of Ophthalmology, Tufts Medical Center, Tufts University School of Medicine, Boston, Massachusetts, United States
  • Arsia Jamali
    Center for Translational Ocular Immunology, Department of Ophthalmology, Tufts Medical Center, Tufts University School of Medicine, Boston, Massachusetts, United States
  • Yashar Seyed-Razavi
    Center for Translational Ocular Immunology, Department of Ophthalmology, Tufts Medical Center, Tufts University School of Medicine, Boston, Massachusetts, United States
  • Brendan Kenyon
    Center for Translational Ocular Immunology, Department of Ophthalmology, Tufts Medical Center, Tufts University School of Medicine, Boston, Massachusetts, United States
    Program in Neuroscience, Tufts University Graduate School of Biomedical Sciences, Boston, Massachusetts, United States
  • Deshea L Harris
    Center for Translational Ocular Immunology, Department of Ophthalmology, Tufts Medical Center, Tufts University School of Medicine, Boston, Massachusetts, United States
  • Driss Zoukhri
    Department of Comprehensive Care, Tufts University School of Dental Medicine, Boston, Massachusetts, United States
  • Pedram Hamrah
    Center for Translational Ocular Immunology, Department of Ophthalmology, Tufts Medical Center, Tufts University School of Medicine, Boston, Massachusetts, United States
    Cornea Service, New England Eye Center; Department of Ophthalmology, Tufts Medical Center, Tufts University School of Medicine, Boston, Massachusetts, United States
  • Footnotes
    Commercial Relationships   Gustavo Ortiz, None; Cecilia Chao, None; Arsia Jamali, None; Yashar Seyed-Razavi, None; Brendan Kenyon, None; Deshea Harris, None; Driss Zoukhri, None; Pedram Hamrah, None
  • Footnotes
    Support  NIH-R01- EY022695, NIH-R21- EY025393, NIH-R01- EY026963, NIH-R01- EY029602 , Tufts Medical Center Institutional Support (PH), Research to Prevent Blindness Challenge Grant and Massachusetts Lions Eye Research Fund, Inc.
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 1961. doi:
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    • Get Citation

      Gustavo Ortiz, Cecilia Chao, Arsia Jamali, Yashar Seyed-Razavi, Brendan Kenyon, Deshea L Harris, Driss Zoukhri, Pedram Hamrah; Effect of Dry Eye Disease on the Kinetics of Lacrimal Gland Dendritic Cells as Visualized by Intravital Multi-Photon Microscopy. Invest. Ophthalmol. Vis. Sci. 2020;61(7):1961.

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

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Abstract

Purpose : Ocular surface dryness induced by desiccating stress (DS) compromises afferent nerve pathways in the lacrimal function unit, which may lead to lacrimal gland (LG) inflammation and reduced tear secretion. We aimed to evaluate the impact of DS-induced DED on conventional dendritic cell (cDC) density, morphology and kinetics in the LG, using intravital multiphoton microscopy (IV-MPM).

Methods : DED was induced by housing 6-8 week old CD11cYFP×Thy1YFP double transgenic mice in a controlled environmental chamber for a period of 2 and 4 weeks (w). DED severity was assessed by corneal fluorescein staining score (CFS) using NEI scale, and tear secretion using phenol red thread test. CD11cYFP cell phenotype and density was quantified by flow cytometry in LG single cell suspensions. A novel approach was developed to image CD11cYFP cells in the LG by IV-MPM, by placing mice onto a custom-designed stage. The LG was exposed carefully and the area to be imaged was covered by a sealed chamber. Analysis of CD11cYFP cell density, 3D morphology, and kinetics was performed using Imaris software. One-way ANOVA with Tukey’s post-hoc was used to assess statistical significance.

Results : DED mice had a 3-fold decreased tear secretion and 12-fold increased fluorescein staining score (p<0.01) compared to naïve mice. IV-MPM and flow cytometry of the LG demonstrated 2-fold increase in the density of cDCs in the LGs of DED mice, compared with controls (p<0.001). cDCs were more spherical in DED at 2w and 4w compared to naïve mice (p<0.001); surface area were found at 2w in DED compared with naïve mice (p<0.001). Mean track speed was 1.8-fold and 1.3-fold higher at 2w and 4w compared to naïve mice (p<0.001). Finally, the meandering index, an index for directionality (scale from 0 to 1), was increased at 4w (0.20±0.01), compared with naïve (0.14±0.01) and 2w (0.15±0.01) of DED (p<0.001).

Conclusions : Our IV-MPM study sheds light into the 3D morphological alterations and cDC kinetics in the LG during DED. While in normal LGs, cDCs exhibit a more dendritic morphology and are less motile, they became more spherical with enhanced motility during DED. This study shows that IV-MPM represents a robust tool to study immune cell trafficking and kinetics in the LG, which might elucidate cellular alterations in immunological diseases, such as DED.

This is a 2020 ARVO Annual Meeting abstract.

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