单疱病毒的耐药及治疗研究

以组织培养和动物实验方法研究3种作用机理不同的抗HSV-1药物的耐受问题,结果HSV对IDU最易耐药,PFA次之,ACV耐药性最差。ACV和PFA耐药株没有交叉耐药性,IDU耐药株对ACV和PFA敏感性稍有下降,但对细胞的感染力未变。ACV治疗IDU耐药株的兔角膜炎有显著疗效,PFA的疗效不明显,IDU治疗可见明显耐药现象。     

抗单纯疱疹病毒单克隆抗体的制备

用单纯疱疹病毒1型(HSV—1)KOS株免疫BALB/C小鼠,取免疫脾细胞与小鼠骨髓瘤细胞SP_2/O融合。经ELISA和免疫荧光(IF)检测,筛选出四株分泌抗HSV型共同性特异性单克隆抗体(McAb)杂交瘤细胞株C_5、B_3,B_8、F_1,其腹水McAbELISA效价分别是10~(-8)、10~(-5)、10~(-5)、10~(-5),而IF则分别为10~(-5)、10~(-4)、10~(-3)、10~(-3)。关键词:McAb,HSV—1 单纯疱疹病毒性角膜炎(HSK)的临床诊断迄今多为臆断。因病毒培养费时且分     

"17997"对无环鸟苷耐药毒株致兔单纯疱疹性角膜炎疗效的观察

目的 :17997为由云南省土壤中分离的一株放线菌产生的广谱抗病毒抗生素 ,在细胞培养内对HSV1、HSV2、HIV1、VZV及CoxB3均有显著抑制活性 ,其体内外抗HSV活性优于无环鸟苷〔1〕。方法 :本实验采用经ACV选择 2 5代产生的ACV耐药株感染兔角膜 ,观察 17997对此毒株致兔角膜炎的疗效 ,行病毒分离 ,计算病毒滴度 ,并与 0 1?V组及赋形剂组比较。结果 :17997对由ACV耐药的HSV1致兔角膜炎有明显疗效 ,减轻角膜病变程度 ,减少排毒数量 ,缩短平均治愈时间。结论 :与ACV相比 ,统计学上有显著性差异 (P <0 0 5 )。     

氨基乙酰无环鸟苷局部治疗实验性单疱角膜-色素膜炎

氨基乙酰无环鸟苷(a'-O-Glycylacyclovir)是无环鸟苷(ACV)的一种水溶性酯衍生物。ACV是一种作用较强和选择性较强的抗单纯疱疹病毒(HSV)1型、2型及水痘带状疱疹病毒药物。细胞培养中,ACV抗HSV-1和HSV-2的作用高于碘苷(IDU)、阿糖腺苷(Ara-A)和三氟胸苷(F_3T)。实验证明3?V眼膏治疗兔HSV-1上皮型角膜炎有效。进一步的临床研究证明3?V眼膏治疗角膜溃疡虽然较用碘苷治疗愈合的快,但其疗效为0.5%或1%IDU眼膏相似,治疗树枝状角膜炎的疗效与Ara-A相同。另外,对IDU或Ara-A耐药的角膜溃疡亦有效。     

HSV对Carbocyclic Oxetanocin G的耐药性及耐药病毒的胸腺嘧啶核苷激酶的活性研究

目的探讨治疗疱疹病毒性角膜炎的新药CarbocyclicoxetanocinG(C.OXTG)耐药性的产生情况及产生耐药性的机制。方法将1型单纯疱疹病毒(HSV1)接种于非洲绿猴肾细胞,在含10μmol/LC.OXTG的环境中连续培养20代,通过空斑抑制试验测定各代病毒株对C.OXTG的敏感性,测定野生HSV1、培养20代后的HSV1的耐药性及其生物克隆株的胸腺嘧啶核苷激酶的活性。结果HSV1在含C.OXTG的环境中连续传代培养4代后,便出现了耐药性。培养20代后的病毒株及其克隆株的ED50为野生株的10倍,且病毒的TK酶活性远远低于野生病毒株。结论HSV1在含C.OXTG的环境中很快产生耐药性且耐药病毒株缺乏TK酶活性。     

无环鸟苷滴眼液点眼的眼内药代动力学研究

无环鸟苷(Acyclorir.ACV)系一选择性抗单纯疱疹病毒(HSV)药物,具有作用强,毒副作用小的优点。眼科配成滴眼液点眼用于治疗各型HSV角膜炎取得良好效果。本文用[~3H]—ACV配成滴眼液(0.1%)给家兔点眼,观察其眼内透性并进行药代动力学分析。     

干扰素联合无环鸟苷抑制单纯疱疹病毒

目的　探讨干扰素 (interferon ,IFN )与无环鸟苷 (acyclovir,ACV)联合使用抑制单纯疱疹病毒 (herpessimplexvirus ,HSV)时的药效及病毒耐药性产生情况。 方法　将HSV Ⅰ分别接种于经IFN α作用 2 4h及未经IFN α处理的非洲绿猴肾细胞中 ,加入不同浓度的ACV ,培养 72h后固定、染色、清点空斑数并计算ACV的半数有效剂量 (ED50 )。将HSV Ⅰ在含ACV及IFN α与ACV的环境中连续培养 10代 ,分别在第 3、5、7和 10代测定ACV的ED50 。结果　ACV对野生HSV Ⅰ的ED50 为 14 2 .0× 10 -9mol·L-1,IFN α与ACV联合用药时ACV对野生HSV Ⅰ的ED50 为 4 7.4× 10 -9mol·L-1。HSV Ⅰ在含ACV的环境中连续培养 7代后即产生了耐药性 ,其ED50 为第 1代的 82倍 ,而HSV Ⅰ在含有ACV和IFN α的环境中培养 10代后未产生耐药性。结论　IFN和ACV联合应用具有协同作用 ,可以减少ACV的用量 ,提高疗效 ;联合用药可以有效延缓病毒对ACV耐药性的产生。     

单纯疱疹病毒侵入受体在单纯疱疹病毒性角膜炎发病机制中的作用

单纯疱疹病毒侵入受体(疱疹病毒侵入介质、连接素-1、连接素-2、3-O-硫酸化的硫酸乙酰肝素等)是单纯疱疹病毒(HSV)侵入细胞及在细胞间扩散所必需的物质.不同的受体对HSV在组织的感染和扩散中具有不同的作用.疱疹病毒侵入介质可介导HSV进入小梁网细胞、结膜上皮细胞和角膜成纤维细胞,连接素则介导HSV侵入神经组织和细胞...     

无环鸟苷治疗单纯疱疹性角膜炎的疗效和对角膜表面的损害作用

目的：观察无环鸟苷（acyclovir,ACV)治疗单纯疱疹性角膜炎（herpes simplex keratitis,HSK)的疗效和对眼角膜上皮损害的毒性作用，以指导临床正确用药。方法：共收集上皮型和浅实质层型HSK患者8例102眼，均分为2组，分别选用1g.L^-1ACV眼液及含1g.L^-1ACV及2g.L^-1玻璃酸钠的正大捷普滴眼液，病例随机分为每天滴眼3次，6次组，用药分为持续10d和15d 2个时间段组，对侧健眼26眼为用药对照组，观察眼刺激症状，睫状充血，角膜溃疡，角膜表面荧光对照组，观察眼刺激症状，睫状充血，角膜溃疡，角膜表面荧光素色，角膜实质层浸润水肿及前房Tyndall现象，治疗前后应用PCR技术检测角膜表面吸取物单纯疱疹病毒（herpes simplex virus ,HSV)变化，结果：用药治疗后角膜表浅溃疡愈合，HSV检测转阴，原角膜树枝状，星芒状及不规则点片状荧光素着色消退（91．8％），继发出现角膜表浅点状着色（78．5％），停止滴药后着色通常2－4d消退，每天滴1g.L^-1ACV眼液3次者较每天滴药6次平均提前2－3d治愈，但后者角膜表面散在点状着色则较前者多见，正大捷普眼液角膜表浅点状损害较滴ACV眼液轻。结论：ACV有抗HSV作用，对表浅HSK的治疗效果好，但长期滴眼可致角膜上皮损害，产生的药物毒性与用药时间正相关。     

单克隆抗体治疗单纯疱疹性角膜炎的临床观察

应用抗HSV 糖蛋白gC 和gD 的单克隆抗体制成的滴眼液,滴眼治疗单纯疱疹病毒性角膜炎患者共43例46只眼,其中治愈39只眼(84.8%),基本治愈7只眼(15.2%),总有效率为100%.表明局部应用单克隆抗体制剂是治疗单纯疱疹病毒性角膜炎的一条新途径。     

无环鸟苷抑制小鼠单纯疱疹性角膜炎潜伏病毒激活作用的研究

目的 证实无环鸟苷能够作用于潜伏单郊病毒的激活阶段,抑制病毒活化。方法 以Ｂａｌｂ／ｃ小鼠作为潜伏感染模型,以紫外线Ｂ照射为激活条件,治疗组给予无环鸟苷口服,停药后１天处死动物检测活化病毒。结果 治疗组３０例中无１例,而对照组３０例中有１２例检测出活化病毒（Ｐ〈０．０１）;治疗至照射后２天的１０只鼠无１例,而相应对照组１０只中６例检出活化病毒（Ｐ〈０．０１）。结论 预防性应用ＡＶＣ特异作用于病毒激活期,有效的抑制病毒活化。     

Novel dipeptide prodrugs of acyclovir for ocular herpes infections: Bioreversion,antiviral activity and transport across rabbit cornea

PURPOSE: A series of dipeptide prodrugs of antiviral nucleoside acyclovir (ACV) were designed to target the oligopeptide transporter on the cornea with an aim of improving the ocular bioavailability and therapeutic activity of ACV. METHODS: Aqueous stability, ocular bioreversion kinetics in various tissues, in vitro antiviral activity, cell proliferation assay and corneal transport characteristics of the dipeptide prodrugs were studied. Results. ACV dipeptide prodrugs were found to be more stable at pH 5.6 in comparison to L-Val-ACV, an amino acid prodrug of ACV. The prodrugs exhibited higher solubility than ACV. Val-Val-ACV and Val-Tyr-ACV were found to have excellent antiviral activity against herpes simplex virus-1 (HSV-1). All the dipeptide prodrugs exhibited lower cytotoxicity as compared to currently approved anti-HSV agent, trifluorothymidine (TFT). Transport of [(3)H] Val-ACV was inhibited significantly in the presence of the dipeptide prodrugs of ACV. Corneal permeabilities of all the ACV dipeptide prodrugs were observed to be higher than ACV possibly due to recognition of the prodrugs by the oligopeptide transporter on the cornea. CONCLUSIONS: The dipeptide prodrugs were found to be more permeable than the parent drug, ACV. More permeable, less cytotoxic ACV dipeptide prodrugs exhibited excellent chemical stability and antiviral activity against herpes simplex virus thereby rendering these lead compounds promising drug candidates against herpes virus infections.     

复方金银花提取液抗Ⅰ型单纯疱疹病毒的实验研究

本文通过细胞培养的方法对复方金银花进行了药物的抗ＨＳＶ１敏感性实验并同无环鸟苷和盐酸吗啉胍进行了对比观察，同时测定了培养液中的乳酸脱氢酶的活力，定量地分析了各种药物抑制ＨＳＶ１对指示细胞的感染和对细胞的保护作用。实验结果表明：复方金银花的抗病毒效果和对靶细胞的保护作用优于无环鸟苷和盐酸吗啉胍，但其作用机理尚有待于进一步研究。     

Contact lens disinfection to prevent transmission of viral disease

Current FDA standards for contact lens disinfecting systems require that viricidal activity be demonstrated against just one strain of herpes simplex virus, type 1. Small and nonenveloped viruses (e.g., adenoviruses) may be more resistant to disinfection than herpes simplex virus (HSV); however, the efficacy of disinfection systems against these other agents is not routinely tested. Currently approved methods of chemical and thermal lens disinfection do appear to be efficient means to inactivate HSV and the human immunodeficiency virus, both of which have lipid envelopes.     

微量地塞米松与抗病毒药物合用对抗单疱病毒作用的影响

报告在组织培养中地塞米松对单纯疱疹病毒(HSV)的生长繁殖无影响,对无环鸟苷(ACV)和环胞苷(CC)的抗病毒作用亦无干扰。在动物实验中,微量地塞米松[0.001%,常用浓度(0.1%)的百分之一]并不恶化上皮型HSV角膜炎。同时证明有效抗病毒药物的合用,微量地塞米松仍保留有良好的消炎作用,能促进上皮型和实质层型单疱角膜炎的痊愈过程。     

Antiviral drug sensitivity in ocular herpes simplex virus infection.

Thirty-nine herpes simplex virus (HSV) isolates were assayed for their sensitivity to 10 different antiviral agents. Of these 39 HSV isolates 10 were cultured from recipient buttons obtained at penetrating keratoplasty in patients with inactive stromal scarring due to recurrent herpetic keratitis, 25 were cultured from patients with conjunctival and ulcerative ocular infections, and the remaining four were laboratory strains with known drug sensitivity patterns, thus providing controls for the experiment. All but one of the 35 clinical isolates of HSV were type 1 and all were sensitive to the 10 antiviral agents. A single type 2 isolate from a young man with recurrent conjunctivitis proved to be resistant to a number of the antiviral agents. Since many of the clinical isolates had been exposed to multiple and protracted antiviral drug treatment, it is suggested that antiviral drug resistance in type 1 HSV ocular infection is not a significant problem.     

Necrotizing Keratitis Caused by Acyclovir-Resistant Herpes Simplex Virus

Background

We report a case of necrotizing keratitis caused by acyclovir (ACV)-resistant herpes simplex virus (HSV) with a clinical appearance similar to a previous fungal keratitis infection.

Methods

Observational case report.

Results

Penetrating keratoplasty was performed in the left eye with a history of herpetic keratitis that resolved with periodic treatment with ACV ointment and a topical steroid. The left eye was painful and red with an abscess and corneal erosion in the peripheral donor cornea. Examination of the scraped corneal epithelium by light microscopy and culturing identified Candida albicans; polymerase chain reaction (PCR) was negative for human herpes viruses. After antifungal treatment, the ocular pain gradually decreased and the lesions slowly improved but recurred with a similar clinical appearance. A second light microscopy examination and cultures were negative for pathogens including C. albicans. PCR was positive for HSV-1 DNA; treatment with 3% topical ACV ointment was unsuccessful. A third examination showed only HSV-1 DNA. Despite antiviral ACV ointment, no clinical improvement occurred based on the HSV DNA copy numbers, which were the same before and after treatment, indicating a possible ACV-resistant strain. When topical trifluorothymidine was substituted for ACV, clinical improvement occurred and the HSV DNA copy numbers decreased.

Conclusion

Necrotizing keratitis induced by ACV-resistant HSV occurred independently after fungal keratitis, with a similar clinical appearance in this case, making diagnosis and treatment difficult. Monitoring the HSV DNA load by real-time PCR could be useful for refractory cases even with atypical clinical appearances.Key words: Herpes simplex virus, Acyclovir-resistant herpes simplex virus, Necrotizing keratitis, Fungal infection, Real-time polymerase chain reaction     

流式细胞技术测定Ⅰ型单纯疱疹病毒的药物敏感性

目的探讨应用流式细胞技术（FMC）测定Ⅰ型单纯疱疹病毒（HSV-1）对抗病毒药物敏感性的可行性。方法分别应用流式细胞技术和空斑减数试验（PRA）测定对无环鸟苷（ACV）敏感的HSV-1病毒株（SM44）和对ACV耐药的病毒株（ACVr）对常用抗病毒药物ACV、更昔洛韦（GCV）、膦甲酸（PFA）和阿糖腺苷（Ara-A）的敏感性。结果两种检测方法均显示ACVr对ACV和GCV的IC50明显高于SM44（P〈0．05），而对PFA和Ara—A的IC50。两株病毒差异无统计学意义（P〉0．05），FCM法和PRA法的结果具有良好的相关性（r=0．9774，P〈0．01）。结论FCM可以用于抗病毒药物敏感性的测定。     

Recurrent herpetic keratitis: failure to detect herpes simplex virus infection using the Syva MicroTrak HSV1/HSV2 direct specimen identification/typing test

A 35-year-old man had developed recurrent herpetic keratitis characterized by dendritic keratitis at intervals of a year. We were able to culture cytopathic agents repeatedly from his lesions by inoculating Vero cells. The cultures yielded definitive evidence of a virus that caused a cytopathic effect within 3 days. However, these virus strains could not be identified as herpes simplex virus (HSV) in immunofluorescence assays using the Syva MicroTrak HSV1/HSV2 direct specimen identification/typing test. Rather they were identified as strains of HSV type 1 (HSV-1) on the basis of plaque morphology, neutralization tests, electron-microscopic examination and DNA restriction endonuclease analysis. Our results allow us to assume the existence of HSV-1 strains isolated clinically that are negative to analysis using the Syva Micro-Trak HSV1/HSV2 direct specimen identification/typing test.     

Effects of anti herpetic drugs on mice with herpetic epithelial keratitis after reactivation of herpes simplex virus type 1

To compare the efficacies of valacyclovir (VCV) and acyclovir (ACV) on murine herpetic epithelial keratitis, mice inoculated with herpes simplex virus type 1(HSV-1) strain McKrae were divided into 6 treatment groups: oral VCV 50 mg/kg and 100 mg/kg, oral ACV 50 mg/kg, ACV eye ointment (EO), ACV eye drops (ED), and placebo. Keratitis scores showed that oral VCV 50 mg/kg, oral ACV, and ACV ED had equivalent efficacies, while oral VCV 100 mg/kg was as efficacious as ACV EO during acute infection. Each treatment group was further divided into the stimulated group with HSV-1 reactivation by immunosuppressant drugs and hyperthermia, and the non-stimulated group without reactivation. We assessed the virus titers in tissues by plaque assay and HSV DNA copy number in the trigeminal ganglia (TG) by real time polymerase chain reaction (PCR). Results showed that the virus titers in the tissues were lowered after reactivation, and the oral VCV group with reactivation had significantly reduced DNA copy number in the TG than the same treatment group without reactivation. In conclusion, oral VCV is as efficacious as ACV EO and significantly suppresses HSV-1 reactivation.