角膜感染后单克隆抗体检测HSV—1研究

单纯疱疹病毒性角膜炎是临床上常见病多发病,占角膜病首位,但目前仍缺乏理想的病因诊断方法。过去有细胞学检查,病毒分离和免疫荧光的检测等,近年来免疫酶技术发展迅     

单纯疱疹病毒Ⅰ型在兔角膜内潜伏和活化的实验研究

在健康新西兰兔角膜上建立HSK模型，待其病变稳定期，一组行自体穿透性角膜移植，另一组与未感染HSV-1的兔角膜行交叉穿透移植，拆线后行肾上腺素离子透入拟使潜伏病毒活化。离子透人后的植片抗原检测均为阳性，透射电镜检查显示病毒感染征象，植片角膜基质细胞培养发生HSV—1自发感染。实验结果显示角膜组织是HSV—1的另一个潜伏感染处和复发源地。     

单纯疱疹病毒Ⅰ型潜伏感染的研究——兔三叉神经节致敏T淋巴细胞检测研究

报告应用抗兔致敏T淋巴细胞的单克隆抗体和ABC免疫组织化学技术对22只实验性单纯疱疹病毒性角膜炎的家兔三叉神经节致敏T淋巴细胞检测的结果，首次揭示在角膜原发感染HSV-I过程中三叉神经节内致敏T淋巴细胞的出现。致敏T淋巴细胞在角膜接种后第13天出现。第85天消失，结果表明：T淋巴细胞在TG内HSV-I潜伏感染的建立过程中起重要作用。本研究对指导临床治疗提供了理论依据。     

单纯疱疹病毒I型潜伏感染的分子生物学研究

单纯疱疹病毒Ｉ型（ＨＳＶ－１）在角膜发生原发感染后可在三叉神经节（ＴＧ）及角膜组织内发生潜伏感染。分子生物学研究发现潜伏期ＴＧ及角膜组织中都存在ＨＳＶ－１的ＤＮＡ片段及特异性ＲＮＡ的转录，它们与ＨＳＶ－１的潜伏和复发有着密切的关系。但ＨＳＶ－１在ＴＧ与角膜内的潜伏机制有所不同，对这一问题的进一步研究将有助于对ＨＳＶ－１潜伏和复发机理的彻底阐明。     

用原位杂交法检测单纯疱疹病毒Ⅰ型在潜伏感染时的少量潜伏相关转录本表达

目的探讨单纯疱疹病毒Ⅰ型在三叉神经节内潜伏感染时的基因表达。方法以 ＢＡＬＢ／ｃ小鼠制备单纯疮疹性角膜炎,种毒后３６天取三叉神经节和角膜,用免疫组化法检测病毒的抗原表达。制备ｃＲＮＡ探针,用原位杂交法检测 ｍ－ＬＡＴ的表达。结果种毒后３６天,免疫组化结果为阴性,原位杂交结果为阳性。结论 ＨＳＶ－１在三叉神经节内潜伏感染时基因的转录并未静止,有ｍ－ＬＡＴ基因的表达。     

Latent infections by herpes simplex virus in experimental animals

Studies in experimental animals, initially the rabbit and more recently the mouse, have been of great importance in establishing present day concepts concerning the phenomenon of herpesvirus latency. These early observations coupled with more recent knowledge of virological consequences following surgical manipulation of the trigeminal tract have led to a general hypothesis for the natural history of herpetic infections: The infection follows a circuit from skin, mucous membrane, or eye (the primary infection) to the corresponding sensory ganglia via associated nerves. Virus becomes latent in the ganglia and later, as a result of one of the many provocations known to be associated with recurrence of herpetic lesions, is reactivated and travels via the nerve to the surface and again produces lesions. Current research investigating this hypothesis is reviewed.     

单纯疱疹病毒角膜感染后的播散

应用细胞培养技术,结合光镜及电镜检查,对小鼠单眼角膜接种HSV-1后病毒在双侧角膜、球后壁、视神经、三叉神经节及脑干的分布进行了动态观察并详细记录。光镜观察炎症逐渐扩展至眼球深层及神经组织,电镜证实细胞中病毒样颗粒。提出HSV-1除通过神经轴浆运输外,还可能通过眼组织及血行播散,解释了某些临床现象,并对控制单疱病毒性眼病的感染播散及防止复发提出了初步建议。     

单纯疱疹病毒眼感染的快速诊断

在兔实验性HSV-Ⅰ眼感染模型上研究角膜刮片ABC法染色检测HSV抗原在HSV眼感染诊断中的价值。结果表明:角膜刮片ABC法染色检测HSV相对于病毒分离(Ⅵ)的敏感性为97.7%,其总体阳性率(68.5%)高于Ⅵ的阳性率(52.9%),尤其在感染两周后更为显著。ABC法可于2小时内完成,操作简便,是一种敏感快速而实用的HSV眼感染诊断方法,值得推广使用。     

Cyclaradine的抗单纯疱疹病毒作用:在体和离体实验研究

目的 探讨(十)—Cyclaradine在体外及动物模型中抗HSV—1及其耐药株的作用。方法 应用VER0细胞培养和实验性单纯疱疹病毒性角膜炎动物模型，观察(十)—Cyclaradine对HSV—1KOS株、F株、HFTC株、dPyK株、7—2667ACVr株和LeBleu株的抑制效果，并与无环鸟苷(acyclovir，ACV)和三氟胸苷(trif1uorothymidine，TFT)作比较。结果 (十)—Cyclaradine在体外及动物模型中有确切的抗HSV—1作用，而且对ACV和TFT的耐药株也同样有效。结论 (十)—Cyclaradine是一种值得进一步研究的治疗单纯疱疹病毒性角膜灸的药物。     

ABC法在单纯疱疹病毒感染检测中的初步应用

对兔单纯疱疹病毒性角膜炎模型眼,用培养24小时联合ABC法染色(C-ABC)快速检测HSV感染,并在HSV-1感染的Vero细胞制成的抗原片上比较了ABC法、PAP法、IIF和IIE检测HSV抗原的敏感性。结果ABC方法检测HSV感染的相对敏感性为95.8%,特异性为100%,其阳性率与单纯培养7天结果接近,显著大于单纯培养24小时的阳性率。抗原片染色敏感性比较表明ABC法较PAP法、IIF及IIE敏感8～64倍。     

17997眼膏对兔实验性单纯疱疹角膜炎的疗效观察

目的　评价不同浓度 17997眼膏治疗实验性单纯疱疹病毒性角膜炎 (herpes sim -plex keratitis,HSK)的疗效及其刺激性。方法　建立兔眼 HSK动物模型 ,给予 5 0、2 0 0 μg·g- 1 17997眼膏滴眼 ,并以 10 g· L- 1无环鸟苷眼膏、赋形剂和空白组作为对照 ,裂隙灯观察角膜病变并评分 ,分离病毒 ,计算病毒滴度评价 17997眼膏疗效。采用 Draize眼刺激实验评价药物毒性。结果　 5 0 μg· g- 1 17997眼膏角膜病变评分及平均病毒滴度均明显增高 ,与赋形剂和空白组统计学上无显著性差异 (P>0 .0 5 )。 2 0 0 μg· g- 1 17997眼膏组与 10 g·L- 1无环鸟苷眼膏对照组角膜病变相似 ,病毒滴度高峰时平均病毒滴度升高不明显 ,2组统计学上无显著性差异 (P>0 .0 5 ) ,与另外 3组统计学上有显著性差异 (P<0 .0 5 )。 17997眼膏的 Draize评分与赋形剂组统计学上无显著性差异 (P>0 .0 5 )。结论　 5 0 μg· g- 1 17997眼膏对 HSK治疗无效 ;2 0 0μg· g- 1 17997眼膏能有效地治疗 HSK,疗效与 10 g· L- 1 无环鸟苷眼膏相似 ,且无眼刺激性     

The role of viral and host genes in corneal infection with herpes simplex virus type 1

Herpes simplex virus infection of the eye is the leading cause of blindness due to infection in the US despite the availability of several antiviral drugs. Studies with animal models have shown that three factors, innate host resistance, the host adaptive immune response, and the strain of virus interact to determine whether an infection is asymptomatic or proceeds to the development of blinding keratitis (HSK). Of these, the role of adaptive immunity has received the most attention. This work has clearly shown that stromal keratitis is an immunopathological disease, most likely due to the induction of a delayed type hypersensitivity response. Substantially less is known about the role of specific host genes in resistance to HSK. The fact that different strains of virus display different disease phenotypes indicates that viral 'virulence' genes are critical. Of the 80 plus HSV genes, few have been formally tested for their role in HSV keratitis. Most studies of virulence genes to date have focused on a single gene or protein and large changes in disease phenotypes are usually measured. Large changes in the ability to cause disease are likely to reduce the fitness of the virus, thus such studies, although useful, do not mimic the natural situation. Viral gene products are known to interact with each other, and with host proteins and these interactions are critical in determining the outcome of infection. In reality, the 'constellation' of genes encoded by each particular strain is critical, and how this constellation of genes works together and with host proteins determines the outcome of an infection. The goal of this review is to discuss the current state of knowledge regarding the role of host and viral genes in HSV keratitis. The roles of specific genes that have been shown to influence keratitis are discussed. Recent data showing that different viral genes cooperate to influence disease severity and confirming that the constellation of genes within a particular strain determines the disease phenotype are also discussed, as are the methods used to test the role of viral genes in virulence. It will become apparent that there is a paucity of information regarding the function of many viral genes in keratitis. Improving our knowledge of the role of viral genes is critical for devising more effective treatments for this disease.     

我国疱疹病毒性角膜炎的十年研究进展

近十年来国内学者对疱疹病毒性角膜炎进行研究,范围涉及疱疹病毒在角膜内潜伏及T细胞和细胞因子的表达在复发机制中的作用、免疫荧光法及PCR法检测病毒的应用、病毒性小梁网炎的临床表现及角膜内皮炎鉴别诊断要点、抗病毒药物的作用与临床选择以及手术治疗学研究等诸多方面.但是目前国内有关此病复发机制的研究较少,有关导致角膜炎的疱疹病毒种类的研究亟待开展.     

The Epidemiology of Herpes Simplex Virus Eye Disease in Northern California

Purpose: To calculate the incidence and prevalence of herpes simplex virus (HSV) eye disease in a large, well-defined population in Northern California, USA, and to determine the recurrence rate following an initial episode of disease in this cohort.

Methods: A retrospective, observational, cohort study using population-based data and medical record review. The patient database of a large, regional health maintenance organization (Northern California Kaiser Permanente) was searched, and the study population consisted of 1,042,351 people over a 1-year study period from 1 July 1998 through 30 June 1999. Only ocular HSV cases with definitive clinical or laboratory confirmed diagnoses were included. Active and inactive cases were included, however only active cases were used in incidence and prevalence calculations. Bilateral disease was counted as one case. Newly diagnosed cases were followed for recurrence from initial presentation through 31 December 2002.

Results: After chart review of 322 possible cases, 71 new cases and 59 previously diagnosed active cases of ocular HSV were confirmed. This resulted in an incidence rate of 6.8 new cases/100,000 person-years (95% confidence interval, CI, 5.3–8.6). Incidence increased with age, and rates were highest in people over 75 years of age (p?<?0.001). The recurrence rate in new cases was 18% for the 3-year follow-up time, and was equal to 5% per year (95% CI 3–9%).

Conclusion: The incidence and prevalence of ocular herpes simplex in this study was lower than previously reported. Incidence increased with age, and there were significantly higher rates in the older population.