单纯疱疹病毒Ⅱ型和人乳头瘤病毒的感染与人工流产术后不孕症的关系

目的探讨性传播病毒和不孕症的关系。方法应用聚合酶链反应（ＰＣＲ）对６０例人工流产术后不孕症妇女和３９例正常妇女进行了生殖道单纯疱疹病毒Ｉ型（ＨＳＶ２）和人乳头瘤病毒（ＨＰＶ）的检测。结果不孕组和对照组ＨＳＶ２的阳性检出率分别是８０．０％和２５．５％，两组间有极显著性差异（Ｐ＜００１）；ＨＰＶ的阳性率分别是５３３％和３３３％，两组间无显著性差异（Ｐ＜０．０５）；ＨＳＶ和ＨＰＶ在两组中的混合感染的阳性率分别是４３．３％（２６／６０）和２３．１％（９／３９），两者有显著性差异（Ｐ＜０．０５）。表明ＨＳＶ２或ＨＳＶ２和ＨＰＶ的混合感染与人工流产术后不孕症有显著的相关性，很可能是不孕的原因之一。两组９９份标志中，ＨＳＶ２和ＨＰＶ混合感染的阳性率为３５．３５％，统计学分析表明，ＨＳＶ２和ＨＰＶ感染与不孕有极显著的相关性χ＝１２．５，Ｐ＜０．０１。结论ＨＳＶ２和ＨＰＶ的感染和不孕症相关     

抗戊型肝炎病毒IgG的检测

应用ＥＬＩＳＡ方法对１９９０年沈阳一起暴发流行的戊型肝炎病人及对照人群７９份血清进行了抗戊型肝炎病毒（ＨＥＶ）ＩｇＧ的检测。显性感染病人与非疫区正常对照人群抗－ＨＥＶＩｇＧ检出率分别为５５．３２％（２６／４７）和２０．００％（２／１０），两者存在显著性差异（ｘ２＝４．１２，Ｐ＜０．０５）。显性感染病人急性期，病后８个月血清抗－ＨＥＶＩｇＧ检出率分别为１００．００％（８／８）和谐４１．９４（１３／３１），前者明显高于后者（ｘ２＝６．８３，Ｐ＜０．０１）。８例病人急性期和病后４个月血清抗体平均滴度分别为０．６８４和０．２３４，抗体滴度下降２．９２３倍，存在显著性差异（Ｔ＝２．２３９，Ｐ＜０．０５）。显性感染病人和ＳＧＰＴ升高无症状人群急性期血清间抗－ＨＥＶＩｇＧ检出率、抗体平均摘度存在明显差异（Ｐ＜０．０５，Ｔ＞１．９６０，Ｐ＜０．０５）。     

CoxsackieB＿3病毒对小鼠高频心电图时域值的作用研究

以Ｂａｌｂ／ｃ小鼠为实验动物，观察了ｃｏｘｓａｃｋｉｅＢ＿３病毒对小鼠高频心电图（ＨＦ－ＥＣＧ）的影响。实验组小鼠腹腔注射０．２ｍｌｃｏｘｓａｃｋｉｅＢ＿３病毒后，分别于第１、３、５、７天记录高频心电图，结果表明：从种毒１天后开始，心率显著增加，分别增加了２４．４％（Ｐ＜０．０１）、１６．３％（Ｐ＜０．０１）、１９．９％（Ｐ＜０．０１）、２９．３％（Ｐ＜０．０１）。从种毒３天后开始，Ｑ－Ｔ间期、Ｔ波时程较种毒前明显延长，Ｑ－Ｔ间期较种毒前分别延长了２４．２％（Ｐ＜０．０１）、１７．５％（Ｐ＜０．０５）、２４．０％（Ｐ＜０．０１）；Ｔ波时程较种毒前分别延长了３０．７％（Ｐ＜０．０１）、２９．１％（Ｐ＜０．０５）、３６．４％（Ｐ＜０．０１）。对照组无此变化。结果提示，病毒可能对窦房结自律细胞的自律性、心室肌细胞的复极化过程产生影响。     

表达单纯疱疹病毒Ⅱ型糖蛋白D的重组痘苗病毒活疫苗株的建立

我们以前曾报道，表达单纯疱疹病毒Ⅱ型糖蛋白Ｄ（ＨＳＶ－２ｇＤ）的重组痘苗病毒（实验疫苗株）能保护被免疫小鼠抵抗致死量ＨＳＶ－２病毒的攻击。在此工作基础上，严格按人用疫苗研究要求的实验条件，成功地建立了表达ＨＳＶ－２ｇＤ的重组痘苗病毒活疫苗株。首先将经聚合酶链反应（ＰＣＲ）修饰的ＨＳＶ－２ｇＤ基因插入痘苗表达质粒ｐＪＳＢ１１７５，置于痘苗病毒Ｐ７５Ｋ早／晚期启动子控制下。将此重组质粒用Ｌｉｐｏｆｅｃｔｉｎ方法转染已受野型ＴＫ＋痘苗病毒天坛７６１株感染的人胚肺二倍体细胞。经同位素探针（３２Ｐ－ＨＳＶ－２ｇＤ）原位杂交法和３轮蚀斑纯化，筛选出基因组内整合有ＨＳＶ－２ｇＤ基因的重组痘苗病毒。斑点和Ｓｏｕｔｈｅｒｎ杂交证实，ＨＳＶ－２ｇＤ基因已插入痘苗病毒基因组内预期的ＴＫ区段，间接免疫荧光检测显示，重组病毒感染细胞后能有效地表达ＨＳＶ－２ｇＤ蛋白。     

聚合酶链反应酶谱分型检测宫颈癌中人乳头瘤病毒 …

目的 探讨人乳头瘤病毒（ＨＰＶ）和单纯疱疹病毒（ＨＳＶ）等对宫颈癌的病因学作用。方法 应用聚合酶链反应（ＰＣＲ）－核酸内切酶分型检测宫颈癌活检组织中ＨＰＶ－ＤＮＡ和ＨＳＶ－ＤＮＡ基因,以正常宫颈组织作对照。结果 在宫颈癌活检细胞中ＨＰＶ－１６,１８型和ＨＳＶ－２型阳性率分别为３８．９％和３４．６％,与正常妇女宫颈组织阳性率均为３．２％比较,差异均有非常显著意义（Ｐ〈０．００１）。结论 ＨＰＶ－１６     

血清乙型肝炎病毒前S1抗原检测及其与病毒复制的关系

用抗Ｓ和抗前Ｓ１单抗的双抗体夹心ＥＬＩＳＡ法检测１５０例慢性乙型肝炎患者、乙型肝炎病毒表面抗原（ＨＢｓＡｇ）携带者和健康人血清中的ＨＢＶ前Ｓ１抗原，其结果和ＨＢＶＤＮＡ聚合酶链反应（ＰＣＲ）、乙型肝炎血清标志的检测结果进行比较。结果表明：前Ｓ１抗原在乙型肝炎病毒ｅ抗原（ＨＢｅＡｇ）阳性组中的检出率和相对滴度显著高于ＨＢｅＡｇ阴性组（Ｐ＜０．０１）；在ＨＢｅＡｇ阴性组中，抗－ＨＢｅ阴性人群前Ｓ１抗原的检出率和相对滴度也显著高于抗－ＨＢｅ阳性人群（Ｐ＜０．０１）。前Ｓ１抗原和ＨＢＶＤＮＡ检测结果的符合率达８０％，两者检出率的相关系数ｒ＝０．９８２６（Ｐ＜０．０１）。结论：血清前Ｓ１抗原和乙型肝炎病毒的存在关系密切。     

人巨细胞病毒感染对肾移植受者血清sIL－2R水平的影响

用ＰＣＲ检测ＨＣＭＶ－ＤＮＡ，ＥＬＩＳＡ法检测ＨＣＭＶ－ＩｇＭ及ＩｇＧ，以诊断肾移植受者ＨＣＭＶ感染。用双抗体夹心法ＥＬＩＳＡ检测６５例肾移植受者血清ｓＩＬ－２Ｒ水平，结果表明：ＨＣＭＶ感染后宿主血清ｓＩＬ－２Ｒ水平明显增高（Ｐ＜０．０１），且ＨＣＭＶ疾病组ｓＩＬ－２Ｒ增高程度大于无症状感染组（Ｐ＜０．０１）；６例原发性ＨＣＭＶ感染者ｓＩＬ－２Ｒ水平与ＩｇＭ水平呈正相关（ｒ＝０．９９０８），提示随感染程度增加，血清ｓＩＬ－２Ｒ水平随之增高，还发现血清ｓＩＬ－２Ｒ水平与Ｃ９４／ＣＤ８比值是负相关（ｒ＝－０．９７８９），说明ＨＣＭｖ感染后ｓＩＬ－２Ｒ水平增高与Ｔ细胞亚群改变有关，反之也说明ｓＩＬ－２Ｒ增高程度可表明体内免疫抑制状态。对于ＨＣＭＶ感染后血清ｓＩＬ－２Ｒ水平增高的机理有待进一步探讨。     

巨细胞病毒感染与可溶性白细胞介素2受体的关系

应用酶联免疫吸附试验（ＥＬＩＳＡ）对１０４例育龄妇女的血清进行了巨细胞病毒（ＨＣＭＶ）ＩｇＧ、ＩｇＭ抗体的检测，同时用ＥＬＩＳＡ双抗体夹心法测定了不同感染状态下血清中可溶性白细胞介素２受体（ｓＩＬ－２Ｒ）的水平，并将ｓＩＬ－２Ｒ水平与未感染ＨＣＭＶ的正常育龄妇女进行了比较。结果，育龄妇女中抗－ＨＣＭＶＩｇＧ的阳性率为８９．４％，ＩｇＭ的阳性率为９．６％，感染ＨＣＭＶ的妇女血清中ｓＩＬ－２Ｒ水平均大于未感染的对照组（１７８．１±５７．３Ｕ／ｍｌ），Ｐ＜０．０５，其中ＩｇＭ阳性者和ＩｇＭ、ＩｇＧ同时阳性者血清中ｓＩＬ－２Ｒ水平最高，分别为９１０±４６５．６Ｕ／ｍｌ和９０５±３４７．８Ｕ／ｍｌ，两者间的差异无显著性意义（Ｐ＞０．０５），但均大于仅抗－ＨＣＭＶＩｇＧ阳性者（４４６．８±１５８．９Ｕ／ｍｌ），Ｐ均＜０．０５。表明，ＨＣＭＶ感染可致ｓＩＬ－２Ｒ水平升高，并且活动性感染者上升明显。提示：ｓＩＬ－２Ｒ可能参与了ＨＣＭＶ的免疫致病机制。     

石家庄地区异常孕产史育龄妇女巨细胞病毒与弓形虫感染研究报告

为了解有异常孕产史妇女（异常组），宫内人巨细胞病毒（ＨＣＭＶ）和弓形虫（ＴＯＸ） 的感染，本文采用ＰＣＲ技术分别对１４５ 例异常组妇女的宫颈分泌物进行了ＨＣＭＶ１０８ 例和ＴＯＸ３７ 例的检测，同时对２７ 例无异常孕产史妇女（ 对照组） 检测了ＨＣＭＶ和ＴＯＸ 结果为异常组ＨＣＭＶ 和ＴＯＸ 的阳性率分别为４４－４４％ 和３２－４％ ，对照组为３－７％ 和０ ％ ，经统计学分析两组间有显著性差异（Ｐ＜ ０－０１） ，结果表明有异常孕产史与ＨＣＭＶ、ＴＯＸ有密切关系进一步证实了孕妇孕前孕中检测ＨＣＭＶ和ＴＯＸ有重要的优生价值。     

病毒激活巨噬细胞产生的一氧化氮与细胞毒的关系

目的：确定新城鸡瘟病毒Ｌｏｄｏｔａ系（ＮＤＶ－Ｌ）能否诱导小鼠腹腔巨细胞（ＰＥＭψ）产生一氧化氮（ＮＯ）以及该 小鼠ＰＥＭψ介导的细胞毒作用是否有依赖关系。方法：采用Ｇｒｉｅｓｓ、ＦＡＣＳ分析和噻唑蓝（ＭＴＴ）方法。结果：ＮＣＶ－Ｌ与ＰＥＭψ任用２Ｈ后就可以稳定地吸附在ＰＥＭψ表面,吸附率达８０％左右;收集ＮＣＶ－Ｌ与ＰＥＭψ作用２４Ｈ上清,发现有ＮＯ的释,释放量与阳性对照组（ＢＸＣＧ－ＬＰＳ－Ｐ     

Acute and latent herpes simplex virus neurological disease in mice immunized with purified virus-specific glycoproteins gB or gD

Groups of 5-week-old BALB/c mice were immunized intraperitoneally with approximately 10 micrograms of purified alum-precipitated glycoprotein gB or gD of either herpes simplex virus type 1 (HSV-1) or type 2 (HSV-2) origin. Control mice received injections of alum-precipitated 1% bovine serum albumin (BSA). Following a second immunization 4 weeks later, seroconversion was confirmed by demonstrating the presence of glycoprotein-specific antibody by immune precipitation. All animals were challenged with lethal doses of either HSV-1 or HSV-2 by footpad inoculation and assessed for acute virus-induced neurological disease and the development of ganglionic latency. Whereas 70% of control (BSA-immunized) HSV-1-infected animals developed ascending myelitis and died, 100% of mice immunized with either gB-1, gB-2, gD-1, or gD-2 antigens remained free of clinical illness and survived HSV-1 challenge. In contrast, gB-1-or gB-2-immunized mice were not protected against acute HSV-2-induced neurological disease and showed a mortality rate of 60-90% (equivalent to that seen in controls), although mean survival times were prolonged. However, significant protection against HSV-2 challenge was observed with gD-1 or gD-2 immunization. When sacral ganglia were removed from surviving mice 9-12 months after virus challenge, latent virus was detected in all gB- or gD-immunized animals, although the extent of latent infection was restricted. These results provide evidence that glycoprotein gD might be superior to glycoprotein gB as an immunogen for the control of acute HSV-1 and HSV-2 neurological disease in mice. However, neither glycoprotein prevents ganglionic latency, the source of virus for recurrent herpesvirus infections.     

Protectivity of herpes simplex virus antigens: Studies in mice on the adjuvant effect of PICLC and on the dependence of protection on T cell competence

The efficacy of a herpes simplex virus type 1 (HSV-1) envelope antigen (EAG) preparation against HSV infection was studied in T cell competent and T cell deficient mice. Immuno-competent mice were successfully protected against herpes simplex virus type 1 (HSV-1) or type 2 (HSV-2) infection when immunized 2 weeks prior to this infection with a heat-inactivated whole virus preparation or a HSV-1 envelope antigen (HSV-1 EAG) preparation. Since HSV-1 EAG was considerably less effective than the whole virus preparation, a polyriboinosinic-polyribocytidylic acid complex with poly-L-lysine and carboxymethylcellulose (PICLC) was used as adjuvant. Immunization with HSV-1 EAG plus PICLC resulted in a pronounced increase of this protection rate as compared with that obtained after immunization solely with HSV-1 EAG. PICLC alone, however, offered no protection when given 2 weeks before challenge. In T cell deficient nu/nu mice no protection was achieved with HSV-1 EAG while their T cell competent, heterozygous littermates were protected. From these results it may be concluded that T cell competence is a prerequisite for establishing a protective immunity against HSV infection after immunization with HSV-1 EAG.     

Herpes simplex virus type 2-mediated disease is reduced in mice lacking RNase L

RNase L helps mediate the antiviral state induced by type I interferons (IFNalphabeta). Although herpes simplex virus (HSV) encodes inhibitors of the IFNalphabeta-induced antiviral response, the IFNalphabeta system serves the body as a first line of defense against HSV. We investigated whether RNase L limits HSV-2 replication and virulence. RNaseL(-/-) and wild-type C57BL/6 mice were infected intravaginally with HSV-2 strain 333. Although initial replication in the genital epithelium was similar, mice lacking RNase L developed less severe genital and neurologic disease than wild-type mice, survived longer, and contained lower viral titers in the nervous system. CD4(+) T cell infiltration into the genital tract and spinal cord of RNase L(-/-) mice was reduced, suggesting that a restricted inflammatory response may account for reduction in disease. Thus, RNase L does not play a significant role in control of HSV-2 infection in vivo; instead, RNase L may regulate aspects of the inflammatory response that contribute to disease.     

Poly(sodium 4-styrene sulfonate): evaluation of a topical microbicide gel against herpes simplex virus type 2 and Chlamydia trachomatis infections in mice

Objective  To investigate the in vivo activity of poly(sodium 4-styrene sulfonate) (T-PSS) gel formulations as topical microbicides.
Methods  The ability of the gel formulations to reduce the incidence of infection when applied prior to pathogen challenge was examined in mouse models of vaginal herpes simplex type 2 (HSV-2) and Chlamydia trachomatis infection, and rectal HSV-2 infection.
Results  In the vaginal HSV-2 challenge studies, 10% T-PSS gel provided significant protection against infection, even when administered 60 min prior to virus challenge ( P  < 0.0001). Both 5% and 10% T-PSS gel formulations significantly reduced the incidence of upper genital tract C. trachomatis infection in animals treated up to 5 min before challenge ( P  < 0.001). However, no protection against C. trachomatis infection was seen in animals treated 30 min before challenge. In mice challenged rectally with HSV-2, both the 5% and 10% T-PSS gels significantly reduced infection at 20 s ( P  < 0.01 for both). However, only the 10% gel provided significant protection when administered 5 min before challenge ( P  < 0.01).
Conclusions  T-PSS gel formulations have promising in vivo activity as topical microbicides.     

反义肿瘤坏死因子α对小鼠眼内单疱病毒感染影响的实验研究

目的观察局部使用肿瘤坏死因子α反义寡核苷酸（TNF-α-ASON）对小鼠眼内单纯疱疹病毒（herpes simplex virus,HSV）-1感染的影响。方法将HSV-1注入小鼠前房建立动物模型。分别于建模前1d,建模后1d和4d结膜下注射TNF-α-ASON和PBS。8d和10d后观察两组小鼠眼部的炎症改变,测定病毒复制和眼内TNF-α含量。结果与对照组相比,8d时实验组葡萄膜炎发病率和眼内TNF-α含量显著降低（P〈0.01）,眼内HSV-1滴度无明显改变;10d时葡萄膜炎发病率和眼内HSV-1滴度都显著升高。结论局部使用TNF-α-ASON可以显著降低HSV-1感染小鼠眼内TNF-α含量,降低视网膜炎发病率。TNF-α在眼内具有抑制HSV-1复制的作用。     

Role of histamine in natural killer cell-dependent protection against herpes simplex virus type 2 infection in mice.

Depletion of natural killer (NK) cells in vivo with anti-NK1.1 monoclonal antibody or anti-asialo-GM1 antiserum drastically reduced survival time in Swiss albino mice infected intravenously (i.v.) with herpes simplex virus type 2 (HSV-2). In contrast, depletion of NK cells did not affect the survival time of mice inoculated with HSV-2 by the intraperitoneal route. A single dose of histamine prolonged survival time in animals inoculated with HSV-2 i.v. but not in animals infected intraperitoneally. Treatment with the histamine H2 receptor antagonist ranitidine alone reduced survival time in i.v.-infected animals and blocked the protective effect of histamine. Histamine or ranitidine did not affect survival time in anti-NK1.1- or anti-asialo-GM1-treated animals. Our data suggest a role for histaminergic mechanisms in NK cell-mediated protection against HSV-2.     

Calcium phosphate nanoparticles induce mucosal immunity and protection against herpes simplex virus type 2

Previously we reported that calcium phosphate nanoparticles (CAP) represented a superior alternative to alum adjuvants in mice immunized with viral protein. Additionally, we showed that CAP was safe and elicited no detectable immunoglobulin E (IgE) response. In this study, we demonstrated that following mucosal delivery of herpes simplex virus type 2 (HSV-2) antigen with CAP, CAP adjuvant enhanced protective systemic and mucosal immunity versus live virus. Mice were immunized intravaginally and intranasally with HSV-2 protein plus CAP adjuvant (HSV-2+CAP), CAP alone, phosphate-buffered saline, or HSV-2 alone. HSV-2+CAP induced HSV-specific mucosal IgA and IgG and concurrently enhanced systemic IgG responses. Our results demonstrate the potency of CAP as a mucosal adjuvant. Furthermore, we show that systemic immunity could be induced via the mucosal route following inoculation with CAP-based vaccine. Moreover, neutralizing antibodies were found in the sera of mice immunized intranasally or intravaginally with HSV-2+CAP. Also, the results of our in vivo experiments indicated that mice vaccinated with HSV-2+CAP were protected against live HSV-2 infection. In conclusion, these preclinical data support the hypothesis that CAP may be an effective mucosal adjuvant that protects against viral infection.     

Vaginal formulations of carrageenan protect mice from herpes simplex virus infection.

The observations from the present study indicate that vaginal formulations of the sulfated polysaccharide carrageenan are highly effective in protecting mice from herpes simplex virus type 2 (HSV-2) infection. Test formulations were placed in the vaginas of progestin-treated mice prior to inoculation with HSV-2. Infection was determined by the presence of inflammation in the genital region and death. At a dose of virus that infected half of the control animals, 1% solutions of either lambda, kappa, or iota carrageenan prevented infection of almost all of the animals. Concentrations as low as 0.05% protected a large majority of the mice. At a dose of virus that infected all of the control mice, 1% solutions of carrageenans protected 85% of the inoculated mice. Other sulfated polysaccharides were less effective or showed no efficacy in preventing HSV-2 infection. These findings suggest that a vaginal formulation of carrageenan may be effective in blocking sexual transmission of HSV-2 in women.     

Double infection of BALB/c mice with Rauscher murine leukemia and Herpes simplex virus type 2

When BALB/c mice were first infected with Rauscher murine leukemia virus (MuLV-R) and then superinfected with Herpes simplex virus type 2 (HSV-2) the inhibition of the evolution of splenomegaly was observed. The effect did not depend on the route of HSV-2 administration. Experiments in which potent anti-interferon serum was administered to double infected mice suggested that the antagonism between MuLV-R and HSV-2 was not mediated by endogenous interferon, HSV-2 was found to replicate in the LPS stimulated spleen cells which are also target cells for MuLV-R; this suggested that the intrinsic interference between both viruses could take place. Mice with Rauscher virus induced disease were found to be more susceptible to infection with Herpesviruses than normal mice.