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Jing Hua, William Stevenson, Thomas Dohlman, Narghes Calcagno, Negar Pirmadjid, Zahra Sadrai, Sunil Chauhan, Daniel Saban, Reza Dana; The CCR7-CCL19/CCL21 Axis Mediates Enhanced Antigen-Presenting Cell Trafficking In High-Risk Corneal Transplantation. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1289.
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High-risk (HR) corneal transplantation is associated with ocular surface inflammation, antigen-presenting cell (APC) maturation and a high rate of graft rejection. Mature APCs are capable of migrating to the draining lymph nodes (LNs) and initiating rejection. This study aimed to examine the magnitude, kinetics and molecular mechanisms of APC trafficking in HR transplantation.
Corneas from congenic CD45.1 or eGFP transgenic mice (C57Bl/6 background) were transplanted to naïve (low risk, LR) or suture-induced neovascularized (HR) BALB/c recipient beds. Ipsilateral draining LNs were excised at 4, 24, and 48h post-transplantation and cells were analyzed using FACS (CD45.1+ and eGFP+ for donor-derived APCs, and YAe+ for alloantigen-bearing recipient APCs). Quantitative PCR and ELISA were performed to evaluate the factors involved in APC homing to draining LNs. Secretory factors from LN explant cultures were assessed using transwell migration assays.
The CD45.1+, eGFP+ and YAe+ cells were detected in the LNs as early as 4h post-transplantation. The frequency of YAe+ cells in HR LNs was significantly higher than in LR at all time points (4h: 0.68±0.14% vs. 0.13±0.06, 24h: 0.70±0.09 vs. 0.32±0.01, and 48h: 0.64±0.06 vs. 0.30±0.05 of total cells, p<0.0001, two-way ANOVA, Bonferroni post-test). CD45.1+ frequencies were significantly higher in HR LNs than in LR LNs at 24h (1.09±0.12% vs. 0.78±0.01 of total cells, p<0.0001) and eGFP+ frequencies showed a similar trend. The donor-derived APCs in HR LNs exhibited higher frequencies of MHC IIhiCCR7+ cells than in LR LNs. CCR7 ligands (CCL19 and CCL21) were significantly upregulated in the HR LNs at both mRNA and protein levels. The supernatant of HR LN explant cultures augmented mature APC migration, and neutralization of CCL19 and CCL21 in transwell assays dramatically reduced migration of mature APCs (0.8x104/well compared to 4.7x104, n=3, p<0.01).
Trafficking of both donor- and recipient-derived APCs is enhanced in HR transplantation. Higher frequencies of mature CCR7+ APCs as a result of enhanced CCL19 and CCL21 expression in the draining LNs of HR recipients augment the trafficking of graft site-derived APCs to the lymphoid tissues.
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