Research review Study and application of monoclonal antibodies against herpes simplex virus

The results of research work completed in recent years in the authors’ laboratory withmonoclonal antibodies (McAbs) against herpes simplex virus (HSV) are reported in this paper as fol-lows.(1) Eighteen hybridoma cell lines steadily producing McAbs against HSV were established byhybridoma technic.(2) The mechanisms of neutralization in vitro and animal protection in vivo medi-ated by different McAbs were investigated.(3) A method of detecting virus antigua was developedwith the McAbs.(4) Tne isolates of virus were typed and antigenically analysed by type-specificand type-common McAbs.(5) HSV antigen in clinical spectimens was detected and directly typed byan ELISA method coating with type-specific and type-common McAb.(6) The target antigens ofMcAbs were identified,purified,and their genes were localized.(7) Experimental rabbitHSV keratitis was treated with McAbs.

DNA vaccine expressing herpes simplex virus 1 glycoprotein C and D protects mice against herpes simplex keratitis

AIM： To investigate whether DNA vaccine encoding herpes simplex virus 1（HSV-1） glycoprotein C（g C） and glycoprotein D（g D） will achieve better protective effect against herpes simplex keratitis（HSK） than DNA vaccine encoding gD alone. METHODS： DNA vaccine expressing gD or gC combined g D（g D.g C） were constructed and carried by chitosan nanoparticle. The expression of fusion protein gD and gC were detected in DNA/nanoparticle transfected 293 T cells by Western-blot. For immunization, mice were inoculated with DNA/nanoparticle for 3 times with 2 wk interval, and two weeks after the final immunization, the specific immune responses and clinical degrees of primary HSK were evaluated. RESULTS： Fusion protein g D.g C could be expressed successfully in cultured 293 T cells. And, p RSC-g C.g DIL21 DNA/chitosan nanoparticle could effectively elicit strongest humoral and cellular immune response in primary HSK mice evidenced by higher levels of specific neutralizing antibody and s Ig A production, enhanced cytotoxicities of splenocytes and nature killer cells（NK）,when compared with those of gD alone or mocked vaccine immunized mice. As a result, gC-based vaccine immunized mice showed least HSK disease. CONCLUSION： gC-based DNA vaccine could effectively prevent the progress of primary HSK, suggesting that this DNA vaccine could be a promising vaccine for HSK treatment in the future.

Corneal esthesiometry and sub-basal nerves morphological changes in herpes simplex virus keratitis/uveitis patients

AIM: To describe and compare corneal sensation and morphological changes of sub-basal corneal nerves by in vivo laser scanning confocal microscopy(LSCM) in herpes simplex virus(HSV) keratitis/uveitis and contralateral, clinically unaffected eyes. METHODS: A prospective clinical study included 30 HSV eyes and 30 contralateral eyes of 30 patients, diagnosed with unilateral HSV keratitis/uveitis. Both eyes underwent a complete ophthalmological examination, Cochet-Bonnet aesthesiometry and LSCM of the central cornea, using the Heidelberg Retina Tomograph III Rostock Cornea Module. After 6 mo, the same examination of the HSV affected and contralateral, clinically unaffected eyes was performed.RESULTS: HSV eyes, as compared to contralateral eyes, demonstrated a significant decrease in mean corneal sensation(3.1±1.6 vs 5.3±0.8 cm), total nerve fibres number(5.7±4.4 vs 15.1±5.4), nerve branches(3.4±3.0 vs 8.4±4.7), main nerve trunks(2.3±1.6 vs 5.8±2.2), and nerve fibres density(7.5±5.6 vs 18.1±5.3 mm/mm2, P<0.05). There was no significant difference between keratitis and uveitis eyes in mean corneal sensation and nerve fibres parameters. After 6 mo, corneal sensation and sub-basal nerve fibres parameters were increased significantly, but did not reach the parameters of contralateral, clinically unaffected eyes.CONCLUSION: Corneal aesthesiometry and LSCM in HSV affected eyes reveals a significant decrease of corneal sensation and sub-basal nerve fibres which recovers at6 mo but does not reach the normal level.

In Vivo Observations and Electron Microscopy of Treatment of Experimental HSV Keratitis with Anti-HSV Monoclonal Antibodies

Purpose: To study the antiviral activity of monoclonal antibodies (McAb) in vivo and identify their effects on experimental herpetic keratitis.Methods: Topical use of anti-HSV monoclonal glycoprotein antibodies was carried out on acute herpetic keratitis of rabbits infected by HSV-1 SM44. The application of the eye drops in each group was five times per day for 14 days by double-blind method. In vivo observation and electron microscopy were performed during the whole procedure. The anti-HSV McAb’s solution was mixed up of five monoclonal antibodies with high neutrilization titers and/or high ADCC activity. Results: Compared with placebo-treated eyes, anti-HSV McAb treatment made statistically significant reduction of herpetic corneal epithelial lesion on rabbits from day 3 to day 14 postinnoculation (P【0. 01). Punctate and short dendritic lesion were the main patterns. The area of involvement was also limited. Electron microscopic analysis showed ultrastructural changes of herpetic corneal infection.

Herpes simplex keratitis:A brief clinical overview

The aim of our minireview is to provide a brief overview of the diagnosis,clinical aspects,treatment options,management,and current literature available regarding herpes simplex keratitis(HSK).This type of corneal viral infection is caused by the herpes simplex virus(HSV),which can affect several tissues,including the cornea.One significant aspect of HSK is its potential to cause recurrent episodes of inflammation and damage to the cornea.After the initial infection,the HSV can establish a latent infection in the trigeminal ganglion,a nerve cluster near the eye.The virus may remain dormant for extended periods.Periodic reactivation of the virus can occur,leading to recurrent episodes of HSK.Factors triggering reactivation include stress,illness,immunosuppression,or trauma.Recurrent episodes can manifest in different clinical patterns,ranging from mild epithelial involvement to more severe stromal or endothelial disease.The severity and frequency of recurrences vary among individuals.Severe cases of HSK,especially those involving the stroma and leading to scarring,can result in vision impairment or even blindness in extreme cases.The cornea's clarity is crucial for good vision,and scarring can compromise this,potentially leading to visual impairment.The management of HSK involves not only treating acute episodes but also implementing long-term strategies to prevent recurrences and attempt repairs of corneal nerve endings via neurotization.Antiviral medications,such as oral Acyclovir or topical Ganciclovir,may be prescribed for prophylaxis.The immune response to the virus can contribute to corneal damage.Inflammation,caused by the body's attempt to control the infection,may inadvertently harm the corneal tissues.Clinicians should be informed about triggers and advised on measures to minimize the risk of reactivation.In summary,the recurrent nature of HSK underscores the importance of both acute and long-term management strategies to preserve corneal health and maintain optimal visual function.

Anti-viral activity of Staphylococcus aureus lysates against herpes simplex virus type-Ⅰinfection:an in vitro and in vivo study

AIM:To investigate the effect of Staphylococcus aureus(S.aures)lysates(SALs)on herpes simplex virus type-Ⅰ(HSV1)infection in human corneal epithelial(HCE)cells and in a mouse model of HSV1 keratitis.METHODS:HCE,Vero,HeLa,and BV2 cells were infected with HSV1[HSV1f strain,HSV1f;HSV-1-H129 with green fluorescent protein(GFP)knock-in,HSV1g].Pre-or post-infection,SAL at various concentrations was added to the culture medium for 24 h.GFP fluorescence in HSV1g or plaque formation by HSV1f were examined.The effects of heat-treated SAL,precooled acetone-precipitated SAL,and SAL subjected to ultrafiltration(100 kDa)were evaluated.The effects of other bacterial components and lysates on HSV1 infection were also tested,including lipoteichoic acid(LTA),peptidoglycan(PGN),staphylococcal protein A(SPA),andα-hemolysin from S.aureus(α-toxin)as well as lysates from a wild-type S.aureus strain,S.epidermidis,and Escherichia coli(W-SAL,SEL,and ECL,respectively).In addition,SAL eye drops were applied topically to BALB/c mice with HSV1 keratitis,followed by in vivo observations.RESULTS:The cytopathic effect,plaque formation(HSV1f),and GFP expression(HSV1g)in infected cells were inhibited by SAL in a dose-dependent manner.The active component of SAL(≥100 kDa)was heat-sensitive and retained activity after acetone precipitation.In HSV1ginfected cells,treatment with LTA-sa,α-toxin,PGN-sa,or SPA did not inhibit GFP expression.SAL,W-SAL,and SEL(but not ECL)decreased GFP expression.In mice with HSV1 keratitis,SAL reduced corneal lesions by 71%.CONCLUSION:The results of this study demonstrate that SAL can be used to inhibit HSV1 infection,particularly keratitis.Further studies are needed to determine the active components and mechanism underlying the effects of SAL.

Systemic IL-1β production as a consequence of corneal HSV-1 infection-contribution to the development of herpes simplex keratitis

This study sought to identify potential therapeutic targets in herpes simplex keratitis(HSK) patients with active and inactive infection by investigating peripheral cytokine production. Peripheral blood mononuclear cells(PBMCs) and serum were prepared from healthy controls and HSK patients during active infection or following treatment(inactive infection). Serum antibody titres were determined by ELISA. Protein expression levels were analysed by Western blot. Cytokine levels were determined by multiplex ELISA. Active corneal herpes simplex virus type 1(HSV-1) infection resulted in significantly elevated peripheral levels of IL-1β in HSK patients compared to healthy controls, and remained significantly increased following treatment. Elevated production of IL-1β in inactive patients was associated with significantly increased levels of IRF3 and STAT1, key proteins involved in promoting anti-viral immune responses. Our data suggest that inflammation persists beyond the period that it is clinically evident and that enhanced peripheral production of IL-1β may have implications for HSV-1 viral clearance in active and inactive HSK patients.

Infectious keratitis in Vietnam:etiology,organisms,and management at Vietnam National Eye Hospital

AIM:To report the etiologies,risk factors,treatments,and outcomes of infectious keratitis(IK)at a major Vietnamese eye hospital.METHODS:This is a retrospective review of all cases of IK at Vietnam National Eye Hospital(VNEH)in Hanoi,Vietnam.Medical histories,demographics,clinical features,microbiological results,and treatment outcomes were reviewed.RESULTS:IK was diagnosed in 1974 eyes of 1952 patients,with ocular trauma being the greatest risk factor for IK(34.2%),frequently resulting from an agriculturerelated injur y(53.3%).The mean duration between symptom onset and presentation to VNEH was 19.3±14.4 d,and 98.7%of patients had been treated with topical antibiotic and/or antifungal agents prior to evaluation at VNEH.Based on smear results of 1706 samples,the most common organisms identified were bacteria(n=1107,64.9%)and fungi(n=1092,64.0%),with identification of both bacteria and fungi in 614(36.0%)eyes.Fifty-five of 374 bacterial cultures(14.7%)and 426 of 838 fungal cultures(50.8%)were positive,with the most commonly cultured pathogens being Pseudomonas aeruginosa,Streptococcus pneumonia,Fusarium spp.,and Aspergillus spp.Corneal perforation and descemetocele developed in 391(19.8%)and 93(4.7%)eyes,respectively.Medical treatment was successful in resolving IK in 50.4%eyes,while 337(17.1%)eyes underwent penetrating or anterior lamellar keratoplasty.Evisceration was performed in 7.1%of eyes,most commonly in the setting of fungal keratitis.CONCLUSION:Ocular trauma is a major risk factor for IK in Vietnam,which is diagnosed in almost 400 patients each year at VNEH.Given this,and as approximately one quarter of the eyes that develop IK require corneal transplantation or evisceration,greater emphasis should be placed on the development of prevention and treatment programs for IK in Vietnam.

Meta-Herpetic Keratitis and Therapeutic Approach: Case Report

The purpose of this study is to disclose the steps in the therapeutic approach for meta-herpetic corneal ulcer. Report of a case with anterior segment photography was used. The 6 months of follow-up results of the case were disclosed. The efficacy of the therapeutic approach for meta-herpetic corneal ulcer was discussed.

Herpes simplex virus-1 infection or Simian virus 40-mediated immortalization of corneal cells causes permanent translocation of NLRP3 to the nuclei

AIM: To investigate into the potential involvement of pyrin containing 3 gene(NLRP3), a member of the nucleotide-binding oligomerization domain-like receptors with cytosolic pattern recognition, in the host defense of corneas against viruses.METHODS: The herpes viral keratitis model was utilized in BALB/c mice with inoculation of herpes simplex virus-1(HSV-1). Corneal tissues removed during therapy of patients with viral keratitis as well as a Simian vacuolating virus 40(SV40)-immortalized human corneal epithelial cell line were also examined.Immunohistochemistry was used to detect NLRP3 in these subjects, focusing on their distribution in tissue or cells. Western blot was used to measure the level of NLRP3 and another two related molecules in NLPR3 inflammasome, namely caspase-1 and IL-1β.RESULTS: The NLRP3 activation induced by HSV-1infection in corneas was accompanied with redistribution of NLRP3 from the cytoplasm to the nucleus in both murine and human corneal epithelial cells. Furthermore,in the SV40-immortalized human corneal epithelial cells,NLRP3 was exclusively located in the nucleus, and treatment of the cells with high concentration of extracellular potassium(known as an inhibitor of NLRP3activation) effectively drove NLRP3 back to the cytoplasm as reflected by both immunohistochemistry and Western blot.· CONCLUSION: It is proposed that herpes virus infection activates and causes redistribution of NLRP3 to nuclei. Whether this NLRP3 translocation occurs with other viral infections and in other cell types merit further study.

抗单纯疱疹病毒1型的IgY制备及其生物学活性检测

目的:制备抗单纯疱疹病毒1型(HSV-1)的卵黄抗体(IgY),探讨该抗体的生物学活性,阐明其抗HSV-1的能力。方法:制备HSV-1灭活疫苗,采用鸡胸多点注射法免疫高产蛋鸡,采用聚乙二醇-6000(PEG-6000)法提纯IgY,采用间接ELISA法、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)技术和蛋白浓度定量试剂盒测定IgY效价、纯度及蛋白水平,采用间接ELISA法检测IgY中和病毒的能力,测定病毒阻断率,以Vero细胞为病毒感染靶细胞,测定不同抗体浓度(0.01560、0.03125、0.06250、0.12500、0.50000、和1.00000 g·L^(-1))作用后细胞病变效应(CPE),根据CPE程度测定IgY体外抗HSV-1能力。结果:制备的IgY效价随免疫时间逐渐升高,达到1/1024000后保持稳定;IgY轻链重链条带清晰;IgY平均蛋白水平为11.544 g·L^(-1);IgY对HSV-1的阻断率可高达77.90%;HSV-1致CPE程度随抗体浓度升高而逐渐降低,当IgY浓度达到0.50000g·L^(-1)及以上时,25%以下细胞(甚至无细胞)发生病变。结论:成功制备出抗HSV-1的IgY,该抗体对HSV-1具有明显的中和阻断能力和体外抑制活性,可用于药物及诊断方法的开发。

继发于单纯疱疹病毒脑炎的抗NMDAR和抗GABA_(BR)双阳性自身免疫性脑炎1例报告及文献复习

目的:分析1例单纯疱疹病毒性脑炎(HSVE)继发抗N-甲基-D-天冬氨酸受体(NMDAR)和抗γ-氨基丁酸B型受体(GABA_(BR))双阳性自身免疫性脑炎(AE)患者的临床表现及诊疗经过,以提高临床医生对该类病的认识。方法:收集1例HSVE继发抗NMDAR和抗GABA_(BR)双阳性AE患者的临床资料,对其诊断和治疗经过进行总结,并结合相关文献进行复习。结果:患者,男性,36岁,以头痛起病,随后出现肢体抽搐,并进展为意识障碍。入院后脑脊液常规生化检测异常,脑脊液单纯疱疹病毒1型(HSV-1) IgG抗体阳性,脑脊液和血清NMDAR抗体检测阳性,头部磁共振成像(MRI)检查提示右侧枕叶白质异常信号,诊断为HSVE继发抗NMDAR脑炎。数月后患者出现精神行为异常、认知障碍和睡眠障碍等症状,血清NMDAR抗体和GABA_(BR)抗体均阳性,诊断为HSVE继发抗NMDAR脑炎和抗GABA_(BR)脑炎。给予激素冲击和静脉注射免疫球蛋白(IVIG)治疗后,患者病情好转出院。随访1年,患者精神症状完全消失,遗留轻度认知功能障碍。结论:HSVE抗病毒治疗有效的恢复期患者临床症状再度恶化时,应高度怀疑继发AE的可能,应尽快完善自身免疫性抗体检测,以期早期诊断,早期治疗,以改善患者预后。

单纯疱疹病毒脑炎后的抗Ri抗体脑炎1例报告

单纯疱疹病毒脑炎是感染性脑炎中最常见的类型,但其预后较差。部分患者会出现复发或者继发自身免疫性脑炎的发生。本文报告1例单纯疱疹病毒脑炎后继发抗Ri抗体脑炎的病例,以帮助提高临床医生对其的认识,避免发生误诊、漏诊等事件发生。尤其在既往有肿瘤的患者中,副肿瘤综合征相关抗体应予以筛查,早期的诊断及免疫治疗对患者的预后起到重要作用。

更昔洛韦眼用凝胶与干扰素滴眼液治疗小儿单疱病毒性角膜炎的效果探究

目的 评定更昔洛韦眼用凝胶与干扰素滴眼液治疗小儿单纯疱疹(单疱)病毒性角膜炎的效果。方法 100例小儿单疱病毒性角膜炎患儿,以计算机随机化方法分为对照组与试验组,每组50例。对照组患儿应用干扰素滴眼液治疗,试验组患儿应用更昔洛韦眼用凝胶联合干扰素滴眼液治疗。比较两组患儿治疗效果,症状消退时间,视力水平,炎性因子指标[C反应蛋白(CRP)、肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)、干扰素-α(IFN-α)]水平及不良反应发生情况。结果 试验组总有效率(98.00%)较对照组(84.00%)更高(P<0.05)。试验组眼部刺激征、溃疡、角膜充血消退时间分别为(6.50±1.22)、(12.51±5.32)、(7.01±1.44)d,均短于对照组的(10.41±2.50)、(21.08±4.65)、(12.38±2.30)d(P<0.05)。治疗后,两组视力、INF-α水平均高于治疗前, CRP、TNF-α、IL-8水平均低于治疗前,且试验组视力(0.95±0.13)、INF-α(118.20±9.83)ng/L较对照组的(0.80±0.12)、(87.64±7.25)ng/L更高, CRP(3.44±0.10)mg/L、TNF-α(29.53±9.20)ng/L、IL-8(49.63±6.05)ng/L较对照组的(8.62±0.75)mg/L、(44.08±8.46)ng/L、(70.22±4.76)ng/L更低(P<0.05)。试验组不良反应发生率(6.00%)与对照组(8.00%)比较差异较小(P>0.05)。结论 小儿单疱病毒性角膜炎患儿接受更昔洛韦眼用凝胶联合干扰素滴眼液方案治疗,能获取较好的效果,缩短康复的时间,更好地改善炎性因子指标水平,且不会增加不良反应的发生几率,意义重大。

单纯疱疹病毒脑炎的临床研究

目的 从临床特点、脑脊液(CSF)特异性抗体检测、神经影像学和治疗等方面对单纯疱疹病毒脑炎(HSE)进行分析,以期加深对本病的认识.提高早期诊断水平和治愈率。方法 在96例病毒性脑炎患者中确诊25例成人HSE病例,并对其临床表现、CT和/或MRI结果、实验室检查、治疗及预后进行分析。结果HSE呈急性或亚急性起病,本组发病年龄中位数为35岁。主要早期表现为发热、头痛、意识和行为人格改变。神经影像学检查有助于诊断。CSF单纯疱疹病毒特异性抗体水平明显升高。意识障碍程度及无环鸟苷的及时应用与预后有关。结论HSE最具特征的早期临床表现为意识障碍和行为人格改变。CT检查不能用于早期诊断。发病1周CT检查正常时,MRI能发现局灶性病变。CSF单纯疱疹病毒特异性抗体水平的升高是重要的诊断依据。无环鸟苷的早期使用是改善预后的关键。

重组人α1b干扰素滴眼液对单纯疱疹病毒性角膜炎的治疗作用

为观察重组人α1b干扰素滴眼液的治疗作用 ,用家兔制作单纯疱疹病毒性角膜炎模型 ,与阿昔洛韦滴眼液进行对照研究。结果显示 ,重组人α1b干扰素滴眼液能缩短单纯疱疹病毒性角膜炎的病程 ,防止角膜出现深层病变 ,改善预后 ,不留瘢痕。研究表明 ,重组人α1b干扰素滴眼液对家兔单纯疱疹病毒角膜炎有一定疗效。

白介素2基因佐剂协同单纯疱疹病毒1型糖蛋白D核酸疫苗免疫诱导的特异性免疫应答

目的研究白介素2(IL-2)cDNA协同单纯疱疹病毒1型(HSV-1)gD核酸疫苗免疫对机体体液免疫和细胞免疫应答的影响,以及在HSV-1病毒角膜攻击时的保护效果。方法利用pcDNA3.1载体分别构建HSV-1糖蛋白D和IL-2的真核表达质粒pgD和pIL-2,体外鉴定其表达。动物实验分为pcDNA3.1空载体对照组、pgD单独免疫组和pgD+pIL-2联合免疫组3组,具体为通过肌肉免疫接种BALB/c小鼠3次,间隔2周,每次接种质粒100μg,第3次免疫后2周,用酶联免疫吸附试验(ELISA)检测抗体滴度并做亚型分析,利用3H-TdR掺入法进行Th细胞增殖实验,ELISA试剂盒检测Th细胞分泌的IL-2、IFN-γ和IL-10水平,耳廓肿胀实验检测迟发型超敏反应(DTH)反应;HSV-1病毒角膜攻击后,裂隙灯显微镜观察角膜上皮病变。结果与pgD单独免疫组相比,pIL-2的协同免疫可以显著提高IgG2a抗体滴度、Th细胞增殖反应和DTH反应,Th细胞分泌IL-2和IFN-γ水平也显著提高,IL-10水平明显下降。在动物保护实验中,pIL-2的协同免疫明显增强了pgD疫苗在病毒攻击时对角膜的保护效果。结论IL-2cDNA的协同免疫可以显著提高pgD核酸疫苗诱导的体液免疫和细胞免疫应答水平,增加核酸疫苗的免疫保护效果。

夏枯草提取物对HSV-Ⅰ及单纯疱疹病毒性角膜炎的作用

目的考察夏枯草提取物对单纯性疱疹病毒性角膜炎的疗效。方法建立单纯性疱疹病毒Ⅰ型(herpes simplex virus-Ⅰ,HSV-Ⅰ)感染Vero细胞的体外模型,以细胞病变效应、治疗指数为观察指标,了解不同浓度夏枯草提取物体外抗病毒效果;采用单纯性疱疹病毒HSV-Ⅰ感染家兔角膜建立体内病毒感染模型,家兔分别用夏枯草提取物、阿昔洛韦、生理盐水治疗15 d,眼角膜每天经荧光素钠染色后,裂隙灯下观察药物对家兔病毒性角膜炎模型的治疗作用。结果体外试验表明夏枯草提取物对感染HSV-Ⅰ病毒的Vero细胞具有明显的抗病毒作用,最大无毒质量浓度TC0为516mg.L-1,治疗指数TI为28.03;体内试验结果显示,与模型组相比,夏枯草提取物和阿昔洛韦组均能有效地治疗单纯疱疹病毒性角膜炎,减轻角膜病变程度,缩短平均治愈时间,其疗效与阿昔洛韦组相似。结论夏枯草提取物对单纯疱疹病毒性角膜炎有显著的疗效,可实施新药研发。

单纯疱疹病毒I型糖蛋白D核酸疫苗的构建及初步研究

目的构建、制备Ⅰ型单纯疱疹病毒糖蛋白Ｄ重组质粒ＤＮＡ疫苗，初步检测其诱导机体产生体液免疫应答的效果，为研制ＨＳＶ－１新型疫苗奠定基础。方法用ＰＣＲ的方法从ＨＳＶ－１病毒基因组中扩增糖蛋白Ｄ（ｇｌｙｃｏｐｒｏｔｅｉｎ Ｄ，ｇＤ）基因，利用基因重组技术构建重组质粒；将重组质粒体外转染真核细胞ＣＯＳ－７，Ｗｅｓｔｅｒｎ ｂｌｏｔｔｉｎｇ检测表达产物；于ＢＡＬＢ／Ｃ小鼠后腿胫前肌注射免疫，０、２周各免疫１次，１００μｇ／次。初次免疫后０、２、４、６周眼眶采血，ＥＬＩＳＡ间接法检测抗体。结果重组质粒酶切出相应大小片段，经测序证实为ＨＳＶ－１ｇＤ序列。Ｗｅｓｔｅｒｎ ｂｌｏｔｔｉｎｇ证实能够在体外真核细胞中表达。免疫小鼠后，产生特异性抗体，抗体滴度１∶２０００。结论ＨＳＶ－１ｇＤ重组质粒ＤＮＡ有可能作为ＨＳＶ－１的ＤＮＡ疫苗，用于防治ＨＳＶ－１感染及其相关疾病。

黄芪对单疱病毒性角膜炎患者Th1/Th2细胞因子的调节作用

目的:探讨黄芪对单疱病毒性角膜炎(herpes simplex keratits,HSK)患者血清Th1/Th2细胞因子水平的影响。方法:选择106例HSK患者随机分为黄芪治疗组和病毒唑治疗组,治疗前后分别采用双抗体夹心法酶联免疫吸附实验(ELISA)检测外周血白细胞介素-2(IL-2),白细胞介素-4(IL-4),白细胞介素-10(IL-10),γ-干扰素(γ-IFN)水平;并以同样方法检测62名体检正常献血员作健康对照组。结果:HSK患者血清中IL-4、IL-10水平较健康对照组显著增高(P<0.01);IL-2;γ-IFN水平较健康对照组显著降低(P<0.01)。黄芪治疗组患者治疗后,血清IL-4、IL-10水平显著降低(P<0.01),而IL-2、γ-IFN水平显著增加(P<0.01)。病毒唑治疗组治疗前后上述指标无明显改变(P>0.05)。结论:黄芪能调节HSK患者体内Th1/Th2细胞因子的失衡状态,改善患者机体免疫功能紊乱状况,这对HSK患者治疗有重要意义。