小鼠角膜移植模型，研究实体器官移植的最常见形式
Summary
Mice have been used as a model for studying many forms of transplantation, including corneal transplantation. We describe in this report a murine model for both acute and late-term corneal transplantation.
Abstract
Corneal transplantation is the most common form of organ transplantation in the United States with between 45,000 and 55,000 procedures performed each year. While several animal models exist for this procedure and mice are the species that is most commonly used. The reasons for using mice are the relative cost of using this species, the existence of many genetically defined strains that allow for the study of immune responses, and the existence of an extensive array of reagents that can be used to further define responses in this species. This model has been used to define factors in the cornea that are responsible for the relative immune privilege status of this tissue that enables corneal allografts to survive acute rejection in the absence of immunosuppressive therapy. It has also been used to define those factors that are most important in rejection of such allografts. Consequently, much of what we know concerning mechanisms of both corneal allograft acceptance and rejection are due to studies using a murine model of corneal transplantation. In addition to describing a model for acute corneal allograft rejection, we also present for the first time a model of late-term corneal allograft rejection.
Introduction
角膜移植是最成功的和常见的类型移植在人执行的一个。为什么进行该手术的原因是损伤，感染性疾病1，或其他形式的非感染性角膜病2的结果。从美国眼库协会的数据显示，46000在2011年进行（见网站：restoresight.org/eye_banks/eye_banks.html）。其成功的标志是有一年不合格率同种异体角膜移植物的范围从10％至15％，并在5年的成功是超过70％3-8。因为许多研究已经显示，角膜移植的成功是直接关系到一个事实，即眼睛的免疫特权部位。负责角膜状态作为免疫特权部位因素包括缺乏既血管和淋巴管在角膜中，相对缺乏抗原呈递细胞的因子，由角膜该suppres产生免疫效应器funtions 9-15，MHC的低表达抗原16，和FasL 17-20的表达。
然而，尽管这些因素诱发这些移植成功的，它们经历排斥3-7。因此，了解这些调解这种排斥反应的机制，以及测试各种治疗，以防止排斥反应是至关重要的。为此，我们在这里描述角膜移植是已经使用了超过20年，研究角膜移植在受控的实验环境的小鼠模型。自移植反应涉及协同工作的许多不同的因素，这将最终确定移植的组织是否失败或成功，则它不可能了解这些因素中的任何体外模型中的重要性。因此，使用完整的动物研究是必需的，以确定哪些因素是重要的成功或者failu重新移植组织。
而其它种类的动物被用来研究角膜移植中，当相比于使用其他种类的鼠模型中具有几个优点。第一个是表示某些转基因的或已被基因靶向至缺乏特异性免疫因子，其功能在移植可以更好地研究了表达小鼠的许多菌株的存在。此外，还有许多试剂（包括重组因子和抗体是中和因子）是特定于小鼠，并且不为其他许多种动物的存在。由于这些因素的存在，该模型已被广泛用于鉴定参与急性角膜移植反应15，17,18,20 -29有关因素。此外，有许多参与角膜移植的因素也是已知的是功能在其他组织中的移植。
Protocol
Representative Results
Discussion
这里描述角膜移植的小鼠模型使研究人员研究人类角膜移植排斥反应的模型，该模型预测的因素是什么，最好既排斥15,17,18,20，26-30和角膜的验收21-25关联移植。不象人类角膜移植，其中给予患者或者局部或全身激素治疗，以任一治疗或预防排斥31，该模型通常用于确定那些是相关的同种异体移植排斥反应在不存在这样的治疗的因素。除了一款型号为急性角膜移植排斥反应?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
The authors would like to thank the many individuals who have worked on and perfected this technique and have been responsible for the generation of many manuscripts both in this lab and others. This work was supported by National Institutes of Health Grant EY12707 (PMS) and an unrestricted grant from Research to Prevent Blindness to Department of Ophthalmology.
Materials
| Name of Material/ Equipment | Company | Catalog Number | Comments/Description |
| Zeiss Surgical Microscope | Zeiss | Rebuilt | |
| 1 mL Syringe | BD | 305122 | |
| 3 mL Syringe | BD | 309657 | |
| 10 mL Syringe | BD | 309602 | |
| Vannus Scissors | Stortz | E-3387 | |
| 11-0 Sutures | Alcon | 717939M | |
| Trephine 2.0mm | Katena | K 2-7520 | |
| Trephine 1.5 mm | Katena | K 2-7510 | |
| Tricaine Hydrochloride 0.5% | Alcon | NDC 0065-0741-12 | |
| Healon | Abbott | Healon OVD | |
| Forceps | FST | 11251-20 | |
| 7-0 Sutures | Alcon | 8065 | |
| 2.5% Phenylephrine HCl | Alcon | NDC 61314-342-02 | |
| 1% Tropicamide | Bausch & Lomb | NDC-24208-585-59 | |
| Hamilton Syringe | Hamilton | 7654-01 | |
| 33 gauge needle | Hamilton | 90033 | |
| Cell Strainer (100 μm nylon) | BD Falcon | 352360 | |
| Hemocytometer | Cardinal Health | B3175 | |
| Trypan Blue | Sigma | T8154 |
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