Arshad Sana, Rana Hafsa, Truong Naomi R, Pattamatta Ushasree, Bertram Kirstie M, White Andrew, Chinnery Holly R, Carnt Nicole A, Cunningham Anthony L
Centre for Virus Research, The Westmead Institute for Medical Research, Westmead, New South Wales, Australia.
Centre for Vision Research, The Westmead Institute for Medical Research, Westmead, New South Wales, Australia.
PLoS Pathog. 2025 May 2;21(5):e1013162. doi: 10.1371/journal.ppat.1013162. eCollection 2025 May.
Herpes Keratitis (HK) is a debilitating infection of the cornea that remains the leading cause of infectious blindness in developed countries. Caused primarily by herpes simplex virus type 1 (HSV-1), it is associated with recurrent inflammation, leading to corneal scarring. This study investigated the initial events during acute HSV-1 infection in the cornea by adapting our human anogenital mucosal explant model to a HSV-1 infected porcine corneal explant model. We infected these corneas topically via high-density microarray patches (HD-MAPs) dipped in GFP-labelled HSV-1. Virus infection and spread was detected by both GFP protein and RNAscope, adapted for HSV-1 DNA. The punctures were consistent, usually in the epithelium but some extended into the underlying stroma. However, HSV-1 was restricted to the corneal epithelium, without spread through the anterior limiting membrane (ALM) or Bowman's layer into the stroma nor to the uppermost epithelial layer. This layer expressed SPRR1A similarly to the stratum granulosum of skin which is refractory to HSV-1 infection. In corneas where infected epithelial cells extended to the ALM, SPRR1A was also observed in this layer, suggesting it may contribute to its barrier function. Such studies of HSV-1 infection and spread will help improve therapy for HK and vaccine design to prevent it.
疱疹性角膜炎（HK）是一种使人衰弱的角膜感染，在发达国家仍是感染性失明的主要原因。它主要由1型单纯疱疹病毒（HSV-1）引起，与反复炎症相关，导致角膜瘢痕形成。本研究通过将我们的人类肛门生殖器黏膜外植体模型应用于HSV-1感染的猪角膜外植体模型，研究了角膜急性HSV-1感染期间的初始事件。我们通过浸入绿色荧光蛋白（GFP）标记的HSV-1的高密度微阵列贴片（HD-MAPs）对这些角膜进行局部感染。通过GFP蛋白和适用于HSV-1 DNA的RNAscope检测病毒感染和传播情况。穿刺点是一致的，通常位于上皮层，但有些延伸到下面的基质层。然而，HSV-1局限于角膜上皮，没有通过前弹力层（ALM）或Bowman层扩散到基质层，也没有扩散到最上层的上皮层。这一层表达的丝聚蛋白前体1A（SPRR1A）与对HSV-1感染具有抵抗力的皮肤颗粒层相似。在感染的上皮细胞延伸到ALM的角膜中，在这一层也观察到了SPRR1A，这表明它可能有助于其屏障功能。此类关于HSV-1感染和传播的研究将有助于改善HK的治疗方法以及预防HK的疫苗设计。
