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Tomas Blanco, Arsia Jamali, Victor G. Sendra, Maria J Lopez, Hamid-Reza Moein, Pedram Hamrah; Plasmacytoid Dendritic Cells in the Mouse Cornea: a Multiphoton Intravital Microscopy Study. Invest. Ophthalmol. Vis. Sci. 2017;58(8):980.
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Plasmacytoid dendritic cells (pDCs) play a main role in linking the innate and adaptive immune responses during inflammation. Herein, we studied dynamic pDCs migratory kinetics in vivo through the use of multiphoton intravital microscopy (MP-IVM)
Corneal inflammation was induced in 8 weeks old DPE-GFP×RAG1-/- transgenic mice (pDC-GFP, n=5/group) by either placement of 3 intrastromal 11-0 nylon sutures in the paracentral cornea, thermal cautery or HSV-1 infection (1x106 PFU McKrae). Naïve pDC-GFP mice served as control. Mice were scanned by a MP-IVM, with MaiTai Ti/Sapphire lasers set at 880nm, at day 7 (suture and HSV-1) or day 3 (cautery). Distribution, kinetics and velocity of pDCs were evaluated and 4D movies rendered using Bitplane software. The meandering index (MI) of directionality (0 maximum, 1 minimum) was calculated. Values from naïve mice (considered static) and inflamed corneas (less than 30 µm of shift) were not included for MI. Results are shown as mean ± SD
Corneal GFP+ pDCs were visualized by MP-IVM. The second harmonic generation delineated pDCs within the anterior stroma, but not in the posterior stroma or epithelium. Naïve corneas show 153±32.55 GFP+ pDCs /mm2 in the periphery and 72.84±11.16 in the center (P=0.036) . In the center, pDC density increased respect to the naïve to 144.10±48.49 cells/mm2 (cautery, P=0.081), 150.40± 61.85 (HSV1, P=0.046) and 315.00± 104.50 (suture, P<0.001). The velocity of pDCs significantly increased from 0.65±0.13 μm/min (naïve) to 3.29 ±1.22 (cautery, P<0.001), 3.67±1.45 (HSV-1, P<0.001) and 4.62±2.03 (suture, P<0.001). The full track length of pDCs in the central cornea significantly increased from 18.62±6.47 µm/hour (naïve) to 127.80±62.22 (cautery, P<0.001), 114.70±54.74 (HSV-1, P<0.001) and 184.4±109.00 (suture, P<0.001). The total displacement length significantly increased from 13.02± 6.47 μm/hour (naïve) to 43.62±23.23 (cautery, P<0.001), 37.11±20.90 (HSV-1, P=0.0023) and (42.19±48.20 (suture, P<0.001). There was no statistical difference in MI between the inflamed groups 0.36±0.15 (cautery), 0.35±0.18 (HSV-1) and 0.37±0.19 (suture)
This study demonstrates, for first time, long-term migratory pDC kinetics in different settings of corneal inflammation through high-resolution MP-IVM. These results suggest that pDCs are active components of the corneal immune system that actively respond to inflammation
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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