Yang Yujia, Flandrin Orneika, Abboud Sara, Jedel Eric, Evans David J, Fleiszig Suzanne M J
Herbert Wertheim School of Optometry & Vision Science, University of California, Berkeley, CA, USA.
Graduate Program in Infectious Diseases and Immunity, University of California, Berkeley, CA USA.
bioRxiv. 2025 Aug 27:2025.08.27.672703. doi: 10.1101/2025.08.27.672703.
Contact lens wear in humans and mice is consistently associated with asymptomatic corneal parainflammation. Here, we tested the hypothesis that the corneal response to lens wear alone can function to protect it against bacterial adhesion enabled by superficial injury. One eye only of mT/mG-LysMcre mice (cell membranes red; Lyz2 cells green) wore a contact lens for 4-6 days. After lens removal, mice were anesthetized and both corneas superficially-injured before bacterial inoculation with either a mouse eyelid commensal () or a corneal pathogen (). Inoculation was repeated hourly for 4 hours under anesthesia before euthanasia. Enucleated eyes were fixed overnight, and adherent bacteria visualized using a universal 16S rRNA-targeted FISH probe () or Blue Fluorescent Protein (). Confocal imaging and Imaris software were used to quantify bacterial adhesion and location in the epithelium, and also the number, location and morphology of Lyz2 cells. For both commensal and , prior lens wear resulted in reduced adhesion to the superficially injured corneas (~ 55% and ~ 45% respectively). In both instances this correlated with increased numbers of corneal Lyz2 cells. Other details differed for the two bacterial types. For the commensal, prior lens wear resulted in bacteria penetrating deeper into the epithelium versus contralateral eyes, with Lyz2 cells extending their processes further into the epithelium and localizing closer to the cornea surface. For , prior lens wear resulted in adherent bacteria closer to the cornea surface, and Lyz2 cells moving further away from it. Moreover, while overall Lyz2 cell sphericity increased for with prior lens wear versus contralateral eyes, it showed no overall change for the commensal. Lyz2 cell volume in the central cornea was decreased for with prior lens wear but increased for the commensal. Thus, corneal responses to prior lens wear can quantitively reduce bacterial adhesion to superficially-injured corneas correlating with a Lyz2 cell response for both a commensal and a pathogen, with differences in details for the two bacterial types. How continued lens wear supersedes this protective response to promote infection pathogenesis remains to be determined, as does the relationship to the Lyz2 cell response.
人类和小鼠佩戴隐形眼镜始终与无症状性角膜副炎症相关。在此，我们检验了以下假设：角膜对单纯佩戴镜片的反应可起到保护作用，防止因表面损伤而导致的细菌黏附。仅mT/mG-LysMcre小鼠（细胞膜呈红色；Lyz2细胞呈绿色）的一只眼睛佩戴隐形眼镜4 - 6天。摘除镜片后，对小鼠进行麻醉，在接种小鼠眼睑共生菌（）或角膜病原体（）之前，对双眼角膜进行表面损伤。在麻醉状态下，每小时重复接种一次，持续4小时，然后实施安乐死。摘除眼球并固定过夜，使用通用的靶向16S rRNA的荧光原位杂交探针（）或蓝色荧光蛋白（）观察黏附细菌。使用共聚焦成像和Imaris软件对细菌在上皮中的黏附及位置进行定量分析，同时也对Lyz2细胞的数量、位置和形态进行分析。对于共生菌和，先前佩戴镜片均导致对表面损伤角膜的黏附减少（分别约为55%和约45%）。在这两种情况下，这都与角膜Lyz2细胞数量增加相关。两种细菌类型的其他细节有所不同。对于共生菌，先前佩戴镜片导致细菌比未佩戴镜片的对侧眼更深地侵入上皮，Lyz2细胞的突起进一步延伸至上皮并更靠近角膜表面定位。对于，先前佩戴镜片导致黏附细菌更靠近角膜表面，而Lyz2细胞则远离角膜表面。此外，与未佩戴镜片的对侧眼相比，先前佩戴镜片的情况下，总体上Lyz2细胞的球形度增加，而对于共生菌则没有总体变化。先前佩戴镜片的情况下，中央角膜中Lyz2细胞的体积对于减少，对于共生菌则增加。因此，角膜对先前佩戴镜片的反应可定量减少细菌对表面损伤角膜的黏附，这与共生菌和病原体的Lyz2细胞反应相关，两种细菌类型在细节上存在差异。持续佩戴镜片如何取代这种保护反应以促进感染发病机制仍有待确定，与Lyz2细胞反应的关系也是如此。
