病毒性皮肤疣小鼠模型的建立

Development of a mouse model of papillomavirus-mediated skin wart

目的疣是因人乳头瘤病毒(HPV)感染导致的皮肤科疾病之一,常表现为寻常疣、尖锐湿疣等症状,治疗后易复发,目前尚无有效的小鼠模型。为研究皮肤疣的防治策略及产品,本研究拟建立病毒性皮肤疣小鼠模型。方法免疫缺陷小鼠麻醉后,利用刀片轻轻刮擦小鼠尾部皮肤制造轻微创口,接种1.5×10^(8) copies小鼠乳头瘤病毒,定期观察尾部皮肤变化,使用不同方法检测病毒DNA、病毒载量及病理分析。结果MmuPV1感染小鼠尾部皮肤16周后,肉眼可见乳头瘤状增生,伴有过度角化以及表面粗糙;HE染色分析为皮肤表面鳞状上皮增生,棘层和颗粒层有空泡变性,可原位检测MmuPV1 E4 RNA转录本。增生部位也可检测到MmuPV1 DNA以及病毒载量。结论本研究利用小鼠乳头瘤病毒MmuPV1感染免疫缺陷小鼠尾部皮肤,成功建立病毒性皮肤疣小鼠模型,支撑皮肤型HPV感染、致病机制等研究。

Objective Verruca is a dermatological disease caused by human papillomavirus(HPV),and includes conditions such as verruca vulgaris and condyloma acuminatum.Although treatments are available to remove skin warts,effective therapies are still lacking.Because of species specificity,the development of animal models of HPV infection has been limited.In this study,mouse papillomavirus(MmuPV1)was used to infect immunodeficient mice to establish a murine model of skin papillomatous hyperplasia.Methods After anesthesia,the tail skin was gently scraped with a blade to produce a slight wound,which was then inoculated with 1.5×10^(8) copies of viral DNA.Changes in the tail skin were observed,viral DNA and load were examined,and pathological analysis was performed.Results Sixteen weeks post-infection with MmuPV1,papillomatous hyperplasia with hyperkeratosis had developed at the infection sites.HE staining showed hyperplasia of squamous epithelium and vacuolar degeneration in the spinous and granular layers.MmuPV1 E4 transcript was detected by RNAscope in situ hybridization.MmuPV1 DNA in the samples was detected by PCR amplification of the E2 gene.Viral DNA was tested in all MmuPV1-infected NU/NU mice samples by qRT-PCR.Conclusions Our result suggest that MmuPV1 skin infection in mice mimics HPV disease,and can therefore serve as a useful model for studying the pathogenesis and natural history of HPV infection.