PCR检测HCMV与HLA的相关性研究

利用聚合酶链反应（ＰＣＲ）法和微量淋巴细胞毒的ＨＩＮ法分别进行了１５７例骨髓移植供受体的人巨细胞病毒（ＨＣＭＶ）和ＨＬＡ－Ａ，Ｂ位点抗原特异性进行了检测，探讨了ＨＣＭＶ感染和ＨＬＡ之间的相关关系，结果显示，在１５７例被测对象中，８５例ＨＣＭＶ感染阳性，７２例阴性，分别统计ＨＣＭＶ阳性组和阴性组的ＨＬＡ－Ａ，Ｂ位点抗原频率，发现ＨＬＡ－Ｂ５在ＨＣＭＶ阳性组中频率较高，而在阴性组中抗原频率较市制是是Ｈ     

丙型肝炎患者血清抗丙型肝炎病毒抗体IgM及HCVRNA检测结果的比较

用酶联免疫吸附试验（ＥＬＩＳＡ）对住院病人抗丙型肝炎病毒抗体（抗－ＨＣＶ）阳性血清标本进行抗－ＨＣＶＩｇＭ的检测，并与ＨＣＶＲＮＡ检测结果比较。结果表明，ＨＣＶＲＮＡ阳性、抗－ＨＣＶ阳性，ＨＣＶＲＮＡ阳性、抗－ＨＣＶ阴性及ＨＣＶＲＮＡ阴性、抗－ＨＣＶ阳性三种类型中均有抗－ＨＣＶＩｇＭ阳性者。结果还表明ＨＣＶＲＮＡ阳性病例的抗－ＨＣＶＩｇＭ阳性率明显高于ＨＣＶＲＮＡ阴性的病例（Ｐ＜０．０５），在临床诊断上ＨＣＶＲＮＡ阳性与阴性病例的肝病大多数为急性肝炎（ＡＨ）和慢性活动性肝炎（ＣＡＨ），ＨＣＶＲＮＡ阳性与阴性比较，各类肝病的病例数无明显差别。     

丙型肝炎患者血清抗丙型肝炎病毒性抗IgM及HCVRN…

用酶联免疫吸附试验对住院病人抗丙型肝炎病毒抗体阳性血清标本进行抗－ＨＣＶＩｇＭ的检测，并与ＨＣＶＲＮＡ检测结果比较。结果表明，ＨＣＶＲＮＡ阳性、抗－ＨＣＶ阳性，ＨＣＶＲＮＡ阳性，抗－ＨＣＶ阴性及ＨＣＶＲＮＡ阴性，抗－ＨＣＶ阳性三类型中均有抗－ＨＣＶＩｇＭ阳性者。     

人巨细胞病毒对小鼠宫颈的诱癌作用

用眼科镊将浸透了病毒母液的明胶海绵块塞入小鼠阴道，直抵宫颈的办法，将紫外线灭活的人巨细胞病毒（ＨＣＭＶ）投给昆明种杂交小鼠。约３１周后，有１５０％小鼠诱发了实验性宫颈癌，另一组加用巴豆油促癌剂，小鼠宫颈癌发率为１８８％，对照组癌发率为０。ＨＣＭＶ诱癌组１２例小鼠宫颈癌组织ＨＣＭＶＩＥ（ｉｍｍｅｄｉａｔｅｅａｒｌｙ）抗原均为阳性，血清ＨＣＭＶＩＥ抗体几何平均滴度为１∶１６７２，对照组以上两个指标均为阴性。ＨＣＭＶ诱癌组小鼠外周血淋巴细胞ＡＮＡＥ阳性率为３８９±６８％，对照组为６７８±８０％。     

HLA多态性与HIV感染及AIDS发病相关性的研究

为研究人类白细胞抗原（ＨＬＡ）和人免疫缺陷病毒（ＨＩＶ）易感性的关系，本文分析比较了以下三组人群的ＨＬＡ表型频率和单倍型频率：①１７２例正常人；②１７例血清ＨＩＶ阴性高危人群；③１８０例血清ＨＩＶ阳性患者，其中２１例发展为艾滋病（ＡＩＤＳ），３７例６个月内ＣＤ４＋淋巴细胞降低至少２０％（ＣＤ４ｄｅｃｌｉｎｅ）。在１７２例对照和１８０例血清ＨＩＶ阳性受试者的比较中发现，其ＨＬＡ表型频率和单倍型频率没有显著差别，提示ＨＩＶ感染与ＨＬＡ无关联。然而，ＨＬＡ－Ｂ２１、ＨＬＡ－Ｂ８、ＨＬＡ－Ｂ３５表型及ＨＬＡ－Ａ１－Ｂ８、ＨＬＡ－Ｂ８－ＤＲ３、ＨＬＡ－Ａ１－Ｂ８－ＤＲ３单倍型与ＣＤ４阳性淋巴细胞下降，或与血清ＨＩＶ感染发展成ＡＩＤＳ病显著相关。１７例ＨＩＶ血清阴性高危人群中，无一携带单倍型ＨＬＡ－Ａ１－Ｂ８－ＤＲ３，提示该单倍型可能与ＨＩＶ感染的抗性相关。     

人巨细胞病毒单克隆抗体的研制检定和应用

采用人巨细胞病毒（ＨＣＭＶ）ＡＤ１６９株作为免疫原，制备出１３株鼠－鼠杂交瘤细胞系。对其中的６株进行了检定．免疫印迹试验结果表明：单克隆抗体（ＭｃＡｂ）７Ｂ４、７Ｄ７、７Ｅ１１、８Ｅ８和８Ｄ６相对应的ＨＣＭＶ多肽分子量分别为４６、１５０、３８、５１７２和６５ｋＤ．ＨＣＭＶ感染人胚肺二倍体细胞（２ＢＳ）后不同时间制成抗原片，与ＭｃＡｂ作间接免疫荧光试验。结果表明：ＭｃＡｂ８Ｂ８相应的病毒多肽为即刻早期抗原，其它５株ＭｃＡｂ相应的病毒多肽均为晚期抗原，６株ＭｃＡｂ等量混合后，标上辣根过氧化物酶，用于ＩｇＭ抗体捕获法ＥＬＩＳＡ（ＭａｃＥＬＩＳＡ）中，并与间接ＥＬＩＳＡ（ＩＥＬＩＳＡ）同时检测ＨＣＭＶ－ＩｇＭ．在未经选择的１００份脐带血中，两法均为阳性的３份，两法均为阴性的９４份；ＭａｃＥＬＩＳＡ阳性而ＩＥＬＩＳＡ阴性的２份血清的特异性试验证明，ＨＣＭＶ－ＩｇＭ确为阳性．ＩＥＬＩＳＡ阳性而ＭａｃＥＬＩＳＡ阴性的１份血清的特异性试验证明，它是由ＲＦ引起的假阳性。     

小儿柯萨奇B组病毒肺炎的临床与体液免疫观察

１４４例临床诊断肺炎的患儿应用酶联免疫吸附试验（ＥＬＩＳＡ）检测柯萨奇Ｂ组病毒（ＣｏｘＢｖｉｒｕｓ，ＣＢＶ）１－６型的感染，检出ＣＢＶ特异性ＩｇＭ阳性２７例，阳性率为１８．７５％，其分型以Ｂ３的阳性率最高，为６２．９０％（１７／２７），以下依次为Ｂ２１８．５０％，Ｂ５１１．１０％，Ｂ４３．７０％，Ｂ３＋Ｂ５３．７０％。ＣＢＶ－ＩｇＭ阳性患儿体液免疫球蛋白ＩｇＡ、ＩｇＧ、ＩｇＭ的检测显示，免疫应答个体反应差异较大，心肌酶谱异常占５９．００％，明显高于阴性组。临床观察病情以轻到中度为主，预防合并症尤为重要。提示ＣＢＶ可引起散发呼吸道感染。     

心脏粘液瘤免疫组织化学观察和组织发生的探讨

对５２例心脏粘液瘤进行免疫组织化学观察。其中有２例伴有腺样结构。免疫标记显示：５２例粘液瘤中全部瘤细胞Ｖｉｍｅｎｔｉｎ为阳性。ＦＶⅢ，Ｄｅｓｍｉｎ和Ａｃｔｉｎ为阴性。５例瘤细胞Ｓ－１００蛋白为阳性。２例伴腺样结构者ＣＥＡ，ＥＭＡ和Ｃｙｔｏｋｅｒａｔｉｏｎ为阳性。组织化学染色显示：腺样结构ＰＡＳ－ＡＢ（ｐＨ１．０），ＰＡＳ－ＡＢ（ｐＨ２．５）和ＨＩＤ－ＡＢ均为阳性。结果提示：心脏粘液瘤可能来源于胚胎     

HBV感染与IgA肾病肾小管间质病变的关系

目的探讨ＩｇＡ肾病ＨＢＶ感染与肾小管间质病变的关系。方法利用原位分子杂交（ＨＢＶＤＮＡ）、免疫组化（ＨＢＡｇ、ＣＤ３、ＣＤ８）以及ＨＢＶＤＮＡＨＢＡｇ和ＨＢＡｇＣＤ４３双标记技术，对９１例ＩｇＡ肾病肾穿刺标本进行研究。结果肾组织内ＨＢＡｇ阳性率为６９．２％。ＨＢＶＤＮＡ原位杂交阳性率为４２９％。ＨＢＶＤＮＡ阳性的病例，双重标记染色发现ＨＢＶＤＮＡ阳性的肾小管上皮细胞可表达ＨＢｃＡｇ或／和ＨＢｓＡｇ。ＨＢＶ感染标记（ＨＢＶＤＮＡ、ＨＢｃＡｇ、ＨＢｓＡｇ）阳性组ＣＤ３阳性细胞和ＣＤ８阳性细胞数明显高于阴性组（Ｐ＜００１），并可见数量不等的Ｔ淋巴细胞入侵ＨＢｃＡｇ及ＨＢｓＡｇ阳性肾小管管壁或围绕其周围。结论感染ＨＢＶ的肾组织细胞能够表达ＨＢＡｇ，并诱导ＣＤ３阳性细胞和ＣＤ８阳性细胞浸润，从而加重肾小管、间质损害。ＨＢＶ感染对ＩｇＡ肾病的发生发展可能起着重要作用     

52 例病毒性肝炎患者肝组织中丙型肝炎病毒 RNA 和 HCAg-NS3 的测定

应用地高辛标记探针原位杂交法和单克隆抗ＨＣＶ－ＮＳ３－ＨＲＰ建立直接酶标免疫组化法分别测定５２例肝炎患者肝组织ＨＣＶＲＮＡ和ＨＣＡｇ－ＮＳ３。结果抗ＨＣＶ阳性组ＨＣＶＲＮＡ检出率５７．１％（１６／２８），ＨＣＡｇ－ＮＳ３检出率５３．６％（１５／２８）；抗ＨＣＶ阴性组其两项检出率均为１２．５％（３／２４）。肝组织中ＨＣＶＲＮＡ阳性物呈蓝紫色细小颗粒存在于肝细胞核或胞浆内，其在肝小叶中的分布可分为３型，即弥漫型、局灶型、散在型。肝组织中ＨＣＡｇ－ＮＳ３阳性物呈棕黄色细小颗粒分布于肝细胞核或胞浆内，以单个或数个阳性细胞散布于肝小叶中。２３例ＨＣＶＲＮＡ或／和ＨＣＡｇ－ＮＳ３阳性病例以肝炎后肝硬化（ＬＣ）病例占多数（１４／２３），其次为慢性重型肝炎（ＣＳＨ）和中度慢性肝炎（ＣＡＨ）。此两种检测方法具有较高符合率（９０．４％，４７／５２），表明病毒核酸及其表达产物均存在于肝细胞内，与ＨＣＶ感染密切相关。这为ＨＣＶ感染诊断提供了直接依据，有利于研究ＨＣＶ感染中病毒复制、慢性化进程、抗病毒治疗监测及重叠感染时病毒相互关系。     

Patterns of human cytomegalovirus infection in term placentas: a preliminary analysis.

BACKGROUND: Primary maternal CMV infection is the major risk factor for symptomatic congenital infection as maternal immunity reduces the risk of transmission to the fetus. Analysis of first trimester placentas showed that virus replicates in the uterus and is transmitted to the placenta causing focal infection. OBJECTIVES AND STUDY DESIGN: We examined 78 term placentas from uncomplicated deliveries for the presence of CMV DNA and evaluated evidence of infection by means of immunohistological and serological analysis. RESULTS: PCR analysis of villus biopsy samples and decidua showed that CMV DNA was present in 62% of tissues. Seven placentas with neutralizing titers were further examined by immunohistology for expression of viral proteins. In placentas with high levels of CMV DNA, fetal blood vessels in the villus core contained neutrophils with viral replication proteins, and macrophages/dendritic cells with glycoprotein B (gB). Cord blood samples from 1 of 11 placentas contained CMV DNA, an indication of replication in the fetal compartment. In placentas with low levels of viral DNA, macrophage/dendritic cells in the villus core contained CMV gB. This pattern was comparable to that seen in early gestation placentas from women with strong neutralizing antibodies. CONCLUSIONS: The results show CMV replication proteins in focal areas of the placenta, implying virus transmission to the fetal circulation. These preliminary results suggest that the incidence of asymptomatic congenital CMV infection might be higher than currently estimated.     

Induction of chromosome aberrations and mitotic arrest by cytomegalovirus in human cells

Human cytomegalovirus (CMV) is potentially an effective but often overlooked genotoxic agent in humans. We report here evidence that indicates that infection by CMV can induce chromosome alterations and mitotic inhibition. The frequency of chromosome aberrations induced was dependent on the input multiplicity of infection (m.o.i.) for human lung fibroblasts (LU), but not for human peripheral blood lymphocytes (PBLs) when both cell types were infected at the GO phase of the cell cycle. The aberrations induced by CMV were mostly chromatid breaks and chromosome pulverizations that resembled prematurely condensed S-phase chromatin. Pulverized chromosomes were not observed in LU cells infected with virus stocks that had been rendered nonlytic by UV-irradiation at 24,000 ergs/mm2 or from infection of human lymphocytes. In LU cells infected with UV-irradiated CMV, the frequency of aberrations induced was inversely dependent on the extent of the exposure of the CMV stock to the UV-light. In permissive CMV infection of proliferating LU cells at 24 hr after subculture, a high percentage (greater than 40%) of the metaphase cells were arrested at their first metaphase and displayed severely condensed chromosomes when harvested 48 hr later. A significant increase (p less than 0.05) in the chromosome aberration frequency was also observed. Our study shows that CMV infection is genotoxic to host cells. The types and extent of damage are dependent on the viral genome expression and on the cell cycle stage of the cells at the time of infection. The possible mechanisms for induction of chromosome damage by CMV are discussed.     

Comparison of In Vitro Lymphocyte Proliferations Induced by Cytomegalovirus-Infected Human Fibroblasts and Cell-Free Cytomegalovirus

In vitro lymphocyte reactivity (LR) to cytomegalovirus (CMV)-infected human fetal fibroblasts (CMVFF) and cell-free CMV were measured by using lymphocytes from healthy donors. Lymphocytes from all seropositive donors were stimulated by CMVFF, whereas lymphocytes from negative donors were not. The optimal stimulator cell-to-lymphocyte ratio was in the range of 1:5 to 1:50, dependent on the virus dose used. LR to cell-free CMV was positive for 15 out of 18 seropositive donors and negative for 14 out of 16 seronegative donors. In most cases LR to CMVFF was considerably higher than LR to cell-free CMV. Within the CMV seropositive group there was no significant correlation between the LR to either CMVFF or cell-free CMV and the levels of antibodies to CMV early antigens or CMV late antigens. There was no strict correlation between LR to CMVFF and to cell-free CMV, especially not in tests with lymphocytes from two patients with CMV mononucleosis. Our data suggest that CMVFF and cell-free CMV are recognized (partly) by different subpopulations of CMV-specific memory lymphocytes. We conclude that the use of CMV-infected cells, in addition to cell-free CMV, in LR tests gives more reproducible and possibly also additional information about CMV-specific cellular immunity.     

Inbred guinea pig model of intrauterine infection with cytomegalovirus.

Outbred guinea pigs have previously been utilized in an experimental model for the study of congenital infection with cytomegalovirus (CMV). Development of an inbred model of intrauterine CMV infection would allow analysis of the cells involved in CMV immunity, studies of transplacental CMV transfer, and investigation of the cellular immune factors that participate in intrauterine CMV infections. This study was therefore designed to assess the inbred guinea pig as a model for the study of congenital CMV infection. Intrauterine fetal and placental infection with CMV was demonstrated in inbred Strain 2 guinea pigs, and the maternal factors influencing transplacental transmission of CMV were evaluated. Infectious virus was recovered from placentas and offspring of mothers that experienced primary CMV infection during pregnancy, but not from placentas and offspring of mothers that were inoculated with CMV prior to pregnancy. However, histologic lesions consisting of focal necrosis and inflammation were seen in tissues of offspring from both groups of mothers. Inoculation of seronegative pregnant Strain 2 animals with low doses of virus (2.5 to 3.5 log10 TCID50) resulted in both placental and fetal CMV infection without significant maternal death. Infection of placentas and offspring occurred in utero regardless of the stage of pregnancy. In addition, infectious virus was detectable in fetal tissues at the time of maternal viremia but also later during the course of maternal infection, ie, 4 weeks after inoculation. These findings indicate that the inbred guinea pig model can be used to investigate the pathogenesis of intrauterine CMV infections.     

艾滋病患者CMV感染致消化道假瘤5例临床病理分析

目的探讨艾滋病(acquired immune deficiency syndrome, AIDS)患者巨细胞病毒(cytomegalovirus, CMV)感染导致消化道假瘤的临床病理特征、诊断与鉴别诊断。方法 5例患者均经内镜活检,采用HE、免疫组化染色,计数CD4阳性细胞,分析AIDS患者CMV感染致消化道假瘤的临床病理特征、免疫表型,并复习相关文献。结果 5例患者临床均表现为溃疡型肿物,镜下未见肿瘤细胞,可见黏膜水肿、坏死、炎症及溃疡形成,其间大量炎性肉芽组织、纤维血管组织增生或肉芽肿形成,导致局部形成肿块或假瘤样表现;可见病毒包涵体,形成"鹰眼"样征象,部分包涵体形态不典型;CMV经免疫组化染色可以更好地显示病毒包涵体。有3例患者外周血CMV-DNA水平增高;5例患者外周血CD4⁺T淋巴细胞计数1~72个/μL;5例患者消化道黏膜CD4⁺T淋巴细胞阳性标记指数为1.53%±1.27%,稍低于AIDS非特异性病变患者(3.6%±0.47%)(P<0.01);显著低于无免疫缺陷患者(27.8%±4.6%)(P<0.01)。结论在AIDS或免疫缺陷患者中出现消化道肿瘤特别是内镜下提示为溃疡型肿瘤,伴血CMV-DNA水平升高(也可正常)及CD4⁺T淋巴细胞明显减少(细胞计数<100个/μL)时,需警惕CMV感染的可能,临床与病理医师应重视内镜下活检,并行HE及免疫组化染色或行原位杂交检测。     

A whole-blood lymphoproliferation assay for measuring cellular immunity against herpes viruses

A whole blood test system was established to study cell-mediated immunity to cytomegalovirus (CMV) and herpes simplex virus (HSV) in a large number of healthy blood donors. Cellular immunity was measured by the in vitro proliferative response (LP) of peripheral lymphocytes. These responded vigorously to several mitogens. Lymphocytes of most individuals responded to HSV, but only a limited number were reactive towards CMV. In parallel, antibodies against CMV and HSV were measured by an ELISA technique. For HSV, good correlation was observed between serological and lymphocyte proliferation results. For CMV, no clear correlation was obtained, only 21 of 40 donors positive in the antibody test being positive in the LP test. The majority of seronegatives were negative in the LP test. Use of virions purified by sucrose gradient centrifugation, or an additional strain of CMV (strain Davis) did not increase the number of donors positive in the LP test. One explanation might be that individuals possessing antibodies against CMV as measured by ELISA but no capacity to react in the LP test had suffered from a CMV infection a long time before, and now showed waning cellular immunity, but antibody still detectable. Use of the whole blood technique on 108 individuals showed that this very simple test works well with various mitogens and at least some antigens.     

无症状HBsAg携带者及其子女转阴血淋巴细胞SCE及DNA损伤修复的研究

目的 观察无症状HBsAg携带者及其子女姐妹染色单体交换(SCE)平均烦率的动态变化以及乙型肝炎病毒(HBV)感染引起的DNA损伤的自然修复。方法 本文采取前瞻性观察,采用20例无症状HBsAg携带者及其8例子女5年后自然转阴外周血淋巴细胞,同时将20例成人无症状HSsAg携带者及其8例携带者子女作为配对组,35例成人和10例儿童HBsAg阴性的健康者作为正常对照组,观察SCE交换频率的动态变化。结果 转阴组SCE均低于成人组和儿童组(P<0.05),而与正常组相比略有增高,但P>0.05。结论 HBV感染引起人类遗传物质损伤后,机体自身对于HBV-DNA修复产生遗传学效应。     

新生儿先天性症状性巨细胞病毒感染与母源性原发及复发CMV 感染的相关性

目的:探讨新生儿先天性症状性巨细胞病毒(Cytomegalovirus,CMV)感染与母源性原发及复发CMV 感染的相关性。方法:选取48 例先天性症状性CMV 感染新生儿及其母亲为感染组,30 例未感染CMV 新生儿及其母亲为阴性组,应用化学发光法(CLIA)检测两组病例外周血特异性抗体IgM/ IgG(CMV-IgM/ IgG)及CMV-IgG 亲合力水平,荧光定量PCR 法检测两组母亲乳汁、新生儿外周血及尿液中CMV-DNA 含量,分析比较其检测结果的差异并回顾性分析比较了两组母亲孕早期CMV-IgG 的浓度水平与本次结果的差异。结果:感染组母亲外周血CMV-IgG 抗体水平及乳汁CMV-DNA 阳性率显著高于对照组,差异有统计学意义(P 均<0.01);病例组患儿与母亲CMV 特异性IgG 抗体比值小于对照组,差异有统计学意义(P 均<0.01);感染组子母间IgG 测定值为负相关性,差异有统计学意义(P<0.01); 阴性组子母间IgG 测定值为正相关性,差异有统计学意义(P<0.01);感染组母亲孕早期CMV-IgG 浓度水平显著低于本次结果,差异有统计学意义(P<0.01);阴性组母亲孕早期CMV-IgG 浓度水平与本次结果无显著性差异(P>0.05)。结论:孕妇体内CMV 活化或再感染导致CMV-IgG 水平升高,是新生儿先天性症状性CMV 感染的高危因素,孕期应关注CMV-IgM/ IgG 动态监测。     

胎儿泌尿系畸形介入性产前诊断

目的探讨胎儿泌尿系畸形的发生与染色体异常、宫内感染的关系.方法在超声介导下对56例泌尿系畸形胎儿抽取脐带血行染色体核型分析,同时采用多聚酶联反应 (polymerase chain reaction,PCR)方法检测TORCH宫内感染.结果①5例胎儿染色体异常,染色体异常率为8.93%,除1例异常核型为单纯泌尿系畸形外,其余4例均合并有其他器官异常;②胎儿脐血检测发现巨细胞病毒(cytomegalovirus,CMV)感染5例,风疹病毒(rubella virus,RV)感染2例,弓形体(toxoplasma,TOX)感染2例,单纯疱疹病毒 (herpes simplex virus,HSV)感染2例,宫内TORCH感染发生率为19.64%.结论染色体异常是引起胎儿泌尿系畸形的重要原因之一,对所有泌尿系统畸形胎儿应行染色体核型分析;而某些宫内感染,尤其是CMV感染,可能引发胎儿泌尿系统畸形.     

Use of recombinant nucleocapsid proteins of the hantaan and nephropathia epidemica serotypes of Hantaviruses as immunodiagnostic antigens

Cytomegalovirus-specific in vitro antibody production (CMV-IVAP) by peripheral blood lymphocytes (PBL) was investigated in 12 renal transplant recipients. CMV-IVAP, CMV-serology, and viral cultures were carried out weekly during at least 8 weeks after transplantation. Nine episodes of CMV infection occurred in eight patients. CMV-IVAP was positive in eight episodes; IgG and/or IgA and/or IgM antibodies reacting with several CMV polypeptides were secreted by PBL. In two patients with primary CMV infection, only IgA antibodies were detected. Cytomegalovirus cultures were positive in eight episodes and serological evidence of CMV infection was obtained in five episodes. In one case, CMV-IVAP was observed before CMV isolation and in another case, before serological evidence of CMV infection. Our study indicates that CMV-IVAP could represent an additional tool for the diagnosis of CMV infection. Our study indicates that CMV-IVAP could represent an additional tool for the diagnosis of CMV infection in renal transplant recipients. © 1993 Wiley-Liss, Inc.