复发性单纯疱疹性角膜基质性角膜炎小鼠模型研究人类HSK
Summary
疱疹性角膜病变的大多数研究使用的主要感染模型。然而，与HSV-1的原发感染通常不导致人类疾病。在这里，我们描述了一个经常性疱疹性角膜病变，更加紧密地模仿人类疾病模型。
Abstract
疱疹性眼部疾病，被称为疱疹性角膜基质炎（HSK），是一种潜在的致盲的角膜感染，超过30万的临床治疗每年访问的结果。 1至2％的临床疾病的患者将体验到失去视力的角膜受感染的。这些案件的绝大多数是由疱疹病毒潜伏感染的激活单纯I型病毒，而不是由于急性疾病。有趣的是，在急性感染是最经常被用来研究这种疾病的模型。然而，有人认为，汉语水平考试将是一个经常性的模型更能反映过程中所发生的临床疾病。经常性的动物模型，为HSK，兔子和老鼠。兔子的优点是，他们的经验重新从潜伏期没有任何已知的刺激。这就是说，它是艰难探索的作用，许多免疫因素在复发性HSK，因为兔模型没有IMMUN，，目的论和遗传资源的鼠标。我们选择使用的小鼠模型，因为它具有复发性HSK的优势，有许多资源，也是我们知道什么时候就会重新激活，因为重新激活诱导暴露于UV-B光。到目前为止，这种模式已使这些实验室用它来定义一些免疫方面的因素，这种疾病是非常重要的。它也使我们能够测试治疗和疫苗的有效性。
Protocol
动物保健注：在这个模型中不使用止痛药，因为它们是抗炎和无效的模型。 1。制备病毒成长HSV-1在Vero细胞（80％汇合），使用0.01多重感染（在T-150烧瓶中，约5×10 5个噬斑形成单位）的起始稀释3-4天，直到所有的细胞都四舍五入容易脱落击中烧瓶中。 删除媒体无菌离心管中，离心SORVALL传奇RT在4°C和3500转每分钟为15分钟。取出后上清液（见1.3），重悬大颗?…
Discussion
这里的单纯疱疹性角膜基质性角膜炎（HSK）中的经常性模型使研究者研究人类的HSK在一个模型与人类疾病的过程中观察到，更符合。因此，模型的优势是疾病发生的背景下的免疫系统，已经在原发病的刺激。由于这些小鼠已经有HSV-1的免疫反应，重新激活，免疫反应的因素，是可能不同于那些最初生成。因此，治疗性干预，以防止初级HSK并不总是会导致复发性HSK的改善，这是我们在各种出版物进?…
Divulgaciones
The authors have nothing to disclose.
Acknowledgements
作者想感谢周杰伦Pepose的教学模型，我们帮助。这项工作是由国家研究院年健康教育资助EY11885（PMS），EY21247（PMS）和医院眼科防盲研究不受限制的资金支持。
Materials
| DMEM | Sigma | D5796 |
| Fetal Bovine Serum | Atlanta Biologicals | S11150 |
| Human Serum | Sigma | S7023 |
| L-Glutamine | Cellgro | 25-005-C1 |
| Pen/ Strep | Sigma | P4333 |
| Fungizone | Invitrogen | 15290018 |
| Centrifuge | Sorvall | Legend RT |
| Centrifuge | Beckman | 52-21 |
| Transilluminatior | UVP | 95-0452-02 |
| Sonicatior | Branson | Sonifier 450 |
| Oak Ridge Tubes | Nalgene | Z 720410 |
| Flasks- 7150 | Corning | 430823 |
| Plates- 12 well | Corning | CLS 3513 |
| Plates- 48 well | Corning | CLS 3548 |
| Sterile Conical Tubes- 30 ml | Corning | CLS 430828 |
| Sterile Cotton-tipped Applicators | Fisher | 23-400-125 |
| 30G Needles | Becton-Dickinson | 305106 |
| 25G Needles | BD | 305122 |
| 10 ml Syringe | BD | 309602 |
| 10 ml Syringe | BD | 309604 |
Referencias
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Citar este artículo
Morris, J., Stuart, P. M., Rogge, M., Potter, C., Gupta, N., Yin, X. Recurrent Herpetic Stromal Keratitis in Mice, a Model for Studying Human HSK. J. Vis. Exp. (70), e4276, doi:10.3791/4276 (2012).