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Thomas Ritter, Oliver Treacy, Aideen Ryan, Mourice Morcos, Marese Cregg, Mikhail Nosov, Lisa O'Flynn; Understanding the Mechanism of Donor Bone Marrow Derived Dendritic Cells in Promoting Corneal Allograft Survival in the Rat. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1290.
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© ARVO (1962-2015); The Authors (2016-present)
To understand the mechanism of ex-vivo generated donor bone marrow derived dendritic cells (BMDCs) on promoting corneal allograft survival in the rat.
BMDCs were propagated from Dark Agouti (DA) rat bone marrow precursor cells in complete medium supplemented with rat GMCSF (5ng/ml) and IL-4 (5ng/ml). For glucocorticoid treatment of BMDCs, dexamethasone (Dexa) (10-6M) was added to the culture. A fully allogeneic rat corneal transplantation model (DA to LEW) was used for in vivo studies. Day 10 donor BMDCs +/- Dexa were harvested and 1x106cells/ml injected intravenously into recipients 7 days prior to corneal transplant surgery. Graft survival and development of opacity, edema and neovascularisation were monitored throughout the therapy. On the average day of rejection the immune microenvironment (cell populations and cytokines expressed) within the graft and the draining lymph nodes was analysed. Alloantibody production was analysed for all experimental groups by flow cytometry.
Ex vivo generated BMDCs have a semi-mature phenotype and can be treated with Dexa to maintain their immature phenotype (reduction in MHC II, CD80 and CD86, n=5 p<0.05). These cells are functional antigen presenting cells but have a reduced allostimulatory capacity compared to control (control DCs: 32.23 ± 2.562%, BMDCs: 4.450 ± 1.354% and Dexa BMDCs: 5.920 ± 0.24%). When applied in vivo BMDC and Dexa BMDC significantly prolong corneal allograft survival (MST> 30d, n=14 p<0.0004 and n=24 p<0.0001 resp.) compared to untreated allogeneic controls (MST 18d, n=11). A significant reduction in the total number of graft infiltrating cells was observed for treated groups (p<0.05 each group n=4). Both treatments resulted in significant ratios of CD4+FoxP3+ cells to CD4+CD25+ cells and increased indoleamine mRNA expression within the graft (p<0.05 each group n=4). The same observation was made at the level of the draining lymph nodes. Interestingly, the alloantibody response (IgG1 and IgG2) was significantly different between BMDCs (n=4) treated and Dexa BMDCs (n=6) treated groups.
Our results demonstrate a significant therapeutic effect of donor-derived BMDCs with and without glucocorticoid treatment in corneal transplant survival. This represents a novel therapeutic approach for the prevention of corneal allograft rejection.
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