Quantification of HSV protein positive neurons and association with an inflammatory cuff post-hyperthermic stress of C57BL/6 mice latently infected with McKrae or 17syn+.

Mice latently infected with McKrae or 17syn+ were subjected to hyperthermic stress and ganglia were harvested at the indicated times post-hyperthermic stress (phs) (n = 5–7 per experiment; results pooled from 2 independent experiments at each time point). HSV proteins were first detected in the whole ganglia and TG were subsequently embedded, cut as 10 μm sections, and counterstained with cresyl violet, as described in Methods. Each HSV protein positive neuron in the sectioned material was then related back to the whole ganglia tissue analysis to confirm the identity as “intact” or “fragmented” and assessed for association or lack of association with an inflammatory cuff, determined as ≥4 inflammatory cell radius (see Fig 6).

将以McKrae株或17syn+株潜伏感染的小鼠施以热应激，于热应激后（post-hyperthermic stress，phs）的指定时间点采集神经节样本；每组实验的样本量为5-7，各时间点的实验结果均合并自2次独立重复实验。首先在全神经节组织中检测到单纯疱疹病毒（Herpes Simplex Virus，HSV）蛋白，随后参照实验方法所述步骤，将三叉神经节（Trigeminal Ganglia，TG）进行包埋、切片（厚度为10 μm），并以克紫（cresyl violet）进行复染色。将切片样本中每一处HSV蛋白阳性神经元与全神经节组织的分析结果进行回溯关联，以确认其为"完整（intact）"或"碎裂（fragmented）"状态，并评估其是否与炎性套存在关联；炎性套的判定标准为炎性细胞半径≥4倍（详见图6）。