Di Zazzo Antonio, Tahvildari Maryam, Subbarayal Brinda, Yin Jia, Dohlman Thomas H, Inomata Takenori, Mashaghi Alireza, Chauhan Sunil K, Dana Reza
1 Schepens Eye Research Institute, Massachusetts Eye & Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, MA.
Transplantation. 2017 Apr;101(4):778-785. doi: 10.1097/TP.0000000000001390.
Corneal neovascularization increases the risk of T cell-mediated allograft rejection. Here, we investigate whether T cells promote angiogenesis in transplantation.
Conventional effector T cells were collected from draining lymph nodes of allogeneic or syngeneic corneal transplanted BALB/c mice. T cells were either cocultured with vascular endothelial cells (VECs) to assess VEC proliferation or used in a mixed lymphocyte reaction assay. Messenger RNA (mRNA) expression of vascular endothelial growth factor (VEGF)-A, -C, and VEGF receptor 2 (VEGF-R2) in VECs was assessed by real-time PCR. VEGF-A protein expression was determined by enzyme-linked immunosorbent assay. Flow cytometry was used to analyze VEGF-R2 expression in corneal CD31 cells, and VEGF-A and IFNγ expression in corneal CD4 T cells.
Allogeneic T cells from high-risk (HR) grafted mice induced more VEC proliferation than those from syngeneic transplant recipients (P = 0.03). Vascular endothelial growth factor-A mRNA and protein expression were higher in T cells from draining lymph nodes (P = 0.03 and P = 0.04, respectively) and cornea (protein; P = 0.04) of HR compared with low-risk (LR) grafted hosts. Vascular endothelial growth factor-A, VEGF-C, and VEGF-R2 mRNA expression were increased in VECs when cocultured with T cells from HR transplants compared with LR transplants and naive mice. In addition, IFNγ blockade in T cell/VEC coculture increased VEC proliferation and VEGF-A protein expression, whereas blocking VEGF-A significantly reduced VEC proliferation (P = 0.04).
Allogeneic T cells from corneal transplant hosts promote VEC proliferation, probably via VEGF-A signaling, whereas IFNγ shows an antiangiogenic effect. Our data suggest that T cells are critical mediators of angiogenesis in transplantation.
角膜新生血管形成增加了T细胞介导的同种异体移植排斥反应的风险。在此,我们研究T细胞是否在移植中促进血管生成。
从同种异体或同基因角膜移植的BALB/c小鼠的引流淋巴结中收集传统效应T细胞。将T细胞与血管内皮细胞(VEC)共培养以评估VEC增殖,或用于混合淋巴细胞反应试验。通过实时PCR评估VEC中血管内皮生长因子(VEGF)-A、-C和VEGF受体2(VEGF-R2)的信使RNA(mRNA)表达。通过酶联免疫吸附测定法测定VEGF-A蛋白表达。流式细胞术用于分析角膜CD31细胞中VEGF-R2表达以及角膜CD4 T细胞中VEGF-A和IFNγ表达。
来自高风险(HR)移植小鼠的同种异体T细胞比来自同基因移植受体的T细胞诱导更多的VEC增殖(P = 0.03)。与低风险(LR)移植宿主相比,HR移植宿主的引流淋巴结(分别为P = 0.03和P = 0.04)和角膜(蛋白;P = 0.04)中的T细胞中血管内皮生长因子-A mRNA和蛋白表达更高。与LR移植和未处理小鼠相比,与来自HR移植的T细胞共培养时,VEC中血管内皮生长因子-A、VEGF-C和VEGF-R2 mRNA表达增加。此外,T细胞/VEC共培养中的IFNγ阻断增加了VEC增殖和VEGF-A蛋白表达,而阻断VEGF-A显著降低了VEC增殖(P = 0.04)。
角膜移植宿主的同种异体T细胞可能通过VEGF-A信号传导促进VEC增殖,而IFNγ显示出抗血管生成作用。我们的数据表明T细胞是移植中血管生成的关键介质。