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K. Maruyama, D. G. Jackson, S. Kinoshita, P. A. D'Amore, J. Stein-Streilein; Differences in Corneal Lymphangiogenesis Between BALB/c and C57BL/6 Mice. Invest. Ophthalmol. Vis. Sci. 2007;48(13):3199.
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The development of lymph vessels (lymphangiogenesis) following corneal transplantation is known to be a risk factor for graft failure. We have previously shown a higher rejection rate for corneal transplantation in C57BL/6 mice than in BALB/c mice. We set out to determine the differences between C57BL/6 and BALB/c mouse corneas that might mediate the different rejection risk.
Flat mounts were prepared from corneas of C57BL/6 and BALB/c mice, and observed by immunofluorescence for the presence of lymph vessels using LYVE-1 as a marker of lymphatic endothelial cells. The innate immune cells were detected using CD11b, CD40, as well as MHC-Class II as a co-stimulatory molecule-marker. Digital images of the flat mounts were taken using a spot image analysis system, and the area covered by lymphatic vessels was measured using NIH Image software. The number of immune cells was counted.
The area of lymph vessels in C57BL/6 corneas was significantly greater than in BALB/c corneas (p=0.03). Moreover, there were significantly more CD11b- (p<0.01) and CD40-, MHC-Class II- (+) cells in the C57BL/6 corneas than in BALB/c mouse corneas. In vitro analysis revealed that CD11b (+) antigen presenting cells express lymphatic endothelial markers and have the capacity to form tube-like structures.
C57BL/6 mouse corneas have more endogenous CD11b (+) cells as well as more lymph vessels. We postulate that the endogenous lymphatic vessels, along with the pro-inflammatory CD11b (+) cells, account for the high risk of corneal graft rejection in C57BL/6 mice. CD11b (+) antigen presenting cells derived from bone marrow cell appear to play an important role in the induction of lymphangiogenesis.
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