Lymphatic vessels growing apart from blood vessels in transplanted corneas after the blockade of vascular endothelial growth factor C

doi:10.3969/j.issn.2095-4344.2016.33.011

Abstract

Abstract:

BACKGROUND: Corneal lymphangiogenesis is beneficial to the transport of corneal antigenic materials, and accelerates the process of antigen presentation, thereby playing an important role in corneal immunity. However, due to the parallel outgrowth of corneal blood and lymphatic vessels in transplanted corneas, it is often difficult to accurately evaluate the role of corneal lymphatic vessels in allograft rejection.
OBJECTIVE: To explore the development of corneal lymphangiogenesis and angiogenesis in transplanted rat corneas after the blockade of vascular endothelial growth factor C (VEGF-C).
METHODS: 130 rats used to establish corneal allogenic transplantation models were equally randomized into two groups: the anti-VEGF-C group and the control group. VEGF-C was blocked in the anti-VEGF-C group by intraperitoneal injection of neutralizing monoclonal anti-VEGF-C antibody every other day for 2 consecutive weeks. Meanwhile, rats in control groups received intraperitoneal injections of saline. Corneal angiogenesis and lymphangiogenesis were characterized using whole mount immunofluorescence, and the immune rejection of the grafts was evaluated by scoring the rejection index (RI). In addition, the expression of VEGF-C was examined by real-time PCR. The relationship of corneal lymphangiogenesis and angiogenesis to RI in transplanted corneas was also characterized.
RESULTS AND CONCLUSION: VEGF-C expression was markedly downregulated after VEGF-C blockade. Corneal lymphangiogenesis developed in parallel with corneal angiogenesis in the control group. While there was a mild reduction in blood vessel area (BVA) and a significant decrease in lymphatic vessel area (LVA) in the anti-VEGF-C group (P < 0.05). In addition, RI was positively correlated with BVA (P < 0.05) and LVA (P < 0.05) in the control group. However, although RI was significantly correlated with BVA (P < 0.05) in the anti-VEGF-C group, the correlation between RI and LVA was not statistically significant (P > 0.05). the graft survival time in the anti-VEGF-C group was significantly increased compared with that in the control group (P < 0.05). Our results show that the outgrowth of lymphatic vessels is separated from that of blood vessels in transplanted corneas by blocking VEGF-C. The blockade of VEGF-C has a significant role in preventing corneal lymphangiogenesis in corneal beds, which results in higher allograft survival rates.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Cornea, Vascular Endothelial Growth Factor C, Tissue Engineering

CLC Number:

R318

Ye Hui, Yan Hao, Zhong Lei, Wang Tao, Deng Juan, Ling Shi-qi. Lymphatic vessels growing apart from blood vessels in transplanted corneas after the blockade of vascular endothelial growth factor C[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(33): 4940-4948.

Figures/Tables 3

References

[1] Maruyama K, Ii M, Cursiefen C, et al. Inflammation- induced lymphangiogenesis in the cornea arises from CD11b-positive macrophages. J Clin Invest. 2005; 115(9):2363-2372.
[2] Cursiefen C, Chen L, Dana MR, et al. Corneal lymphangiogenesis: evidence, mechanisms, and implications for corneal transplant immunology. Cornea. 2003;22(3):273-281.
[3] Yamagami S, Dana MR. The critical role of lymph nodes in corneal alloimmunization and graft rejection. Invest Ophthalmol Vis Sci. 2001;42(6):1293-1298.
[4] Yamagami S, Dana MR, Tsuru T. Draining lymph nodes play an essential role in alloimmunity generated in response to high-risk corneal transplantation. Cornea. 2002;21(4):405-409.
[5] Ling S, Qi C, Li W, et al. Crucial role of corneal lymphangiogenesis for allograft rejection in alkali-burned cornea bed. Clin Experiment Ophthalmol. 2009;37(9):874-883.
[6] Dietrich T, Bock F, Yuen D, et al. Cutting edge: lymphatic vessels, not blood vessels, primarily mediate immune rejections after transplantation. J Immunol. 2010;184(2):535-539.
[7] Ling SQ, Liu C, Li WH, et al. Corneal lymphangiogenesis correlates closely with hemangiogenesis after keratoplasty. Int J Ophthalmol. 2010;3(1):76-79.
[8] Cursiefen C, Schlötzer-Schrehardt U, Küchle M, et al. Lymphatic vessels in vascularized human corneas: immunohistochemical investigation using LYVE-1 and podoplanin. Invest Ophthalmol Vis Sci. 2002;43(7): 2127-2135.
[9] Liu YC, Peng Y, Lwin NC, et al. A biodegradable, sustained-released, prednisolone acetate microfilm drug delivery system effectively prolongs corneal allograft survival in the rat keratoplasty model. PLoS One. 2013;8(8):e70419.
[10] Chang LK, Garcia-Cardeña G, Farnebo F, et al. Dose-dependent response of FGF-2 for lymphangiogenesis. Proc Natl Acad Sci U S A. 2004; 101(32):11658-11663.
[11] Chang L, Kaipainen A, Folkman J. Lymphangiogenesis new mechanisms. Ann N Y Acad Sci. 2002;979:111-119.
[12] Holland EJ, Olsen TW, Sterrer J, et al. Suppression of graft rejection using 15-deoxyspergualin in the allogeneic rat penetrating keratoplasty model. Cornea. 1994;13(1):28-32.
[13] Zhang W, Pan Z, Zhai C. Efficacy of topical cyclosporine A on keratoplasty rejection in rats. Zhonghua Yan Ke Za Zhi. 2001;37(2):140-143.
[14] Yuen D, Grimaldo S, Sessa R, et al. Role of angiopoietin-2 in corneal lymphangiogenesis. Invest Ophthalmol Vis Sci. 2014;55(5):3320-3327.
[15] Tang X, Sun J, Du L, et al. Neuropilin-2 contributes to LPS-induced corneal inflammatory lymphangiogenesis. Exp Eye Res. 2016;143:110-119.
[16] Ling S, Lin H, Liang L, et al. Development of new lymphatic vessels in alkali-burned corneas. Acta Ophthalmol. 2009;87(3):315-322.
[17] Mouta C, Heroult M. Inflammatory triggers of lymphangiogenesis. Lymphat Res Biol. 2003;1(3):201-218.
[18] Yan H, Qi C, Ling S, et al. Lymphatic vessels correlate closely with inflammation index in alkali burned cornea. Curr Eye Res. 2010;35(8):685-697.
[19] Yan H, Yuan J, Peng R, et al. The Blockade of Vascular Endothelial Growth Factor C Effectively Inhibits Corneal Lymphangiogenesis and Promotes Allograft Survival. J Ocul Pharmacol Ther. 2015;31(9):546-554.
[20] Oh SJ, Jeltsch MM, Birkenhäger R, et al. VEGF and VEGF-C: specific induction of angiogenesis and lymphangiogenesis in the differentiated avian chorioallantoic membrane. Dev Biol. 1997;188(1):96-109.
[21] Skobe M, Hawighorst T, Jackson DG, et al. Induction of tumor lymphangiogenesis by VEGF-C promotes breast cancer metastasis. Nat Med. 2001;7(2):192-198.
[22] Jeltsch M, Kaipainen A, Joukov V, et al. Hyperplasia of lymphatic vessels in VEGF-C transgenic mice. Science. 1997;276(5317):1423-1425.
[23] Mäkinen T, Jussila L, Veikkola T, et al. Inhibition of lymphangiogenesis with resulting lymphedema in transgenic mice expressing soluble VEGF receptor-3. Nat Med. 2001;7(2):199-205.
[24] Ling S, Qi C, Li W, et al. The expression of vascular endothelial growth factor C in transplanted corneas. Curr Eye Res. 2009;34(7):553-561.
[25] Oh SJ, Jeltsch MM, Birkenhäger R, et al. VEGF and VEGF-C: specific induction of angiogenesis and lymphangiogenesis in the differentiated avian chorioallantoic membrane. Dev Biol. 1997;188(1):96-109.
[26] Kriehuber E, Breiteneder-Geleff S, Groeger M, et al. Isolation and characterization of dermal lymphatic and blood endothelial cells reveal stable and functionally specialized cell lineages. J Exp Med. 2001;194(6):797-808.
[27] Lymboussaki A, Achen MG, Stacker SA, et al. Growth factors regulating lymphatic vessels. Curr Top Microbiol Immunol. 2000;251:75-82.
[28] Cao Y, Linden P, Farnebo J, et al. Vascular endothelial growth factor C induces angiogenesis in vivo. Proc Natl Acad Sci U S A. 1998;95(24):14389-14394.