Correlates of placental infection with cytomegalovirus, parvovirus B19 or human herpes virus 7

Vertical transmission of viruses is an important cause of morbidity in the fetus and neonate. Placental viral infection indicates risk of vertical transmission, but not always transmission to, or disease of the fetus. Specimens from mothers and babies from three groups-two prospective and one retrospective cohort-were tested for pathogens of teratogenic potential using multiplex PCR. Placental infection was present in 13% of the 105 samples collected. Assessment of the prospective cohorts showed cytomegalovirus (CMV) detected in 4% of placentae from unselected women, parvovirus B19 in 1% and Ureaplasma parvum in 1% of placentae. In a retrospective cohort of women at high risk of transmitting congenital infection due to seroconversion during pregnancy, miscarriage or stillbirth, CMV was detected in 64% and human herpes virus type 7 in 9% of placentae. Of 14 PCR-positive placentae, two were associated with the birth of a living symptomatic infant, two with stillbirth, one with miscarriage, and two with elective terminations of pregnancy. Directed laboratory assessment of women at high risk of transmitting congenital infection, on the basis of clinical or laboratory markers, is important for accurate diagnosis of adverse outcomes of pregnancy. However, routine screening for viruses in the placentae from women with a low-risk serological profile for transmitting congenital infection is unlikely to result in significant numbers of additional diagnoses and is confounded by inadequacy of current diagnostic methods. The major pathogen detected in all cases of placental infection associated with fetal death was human CMV.     

Antibodies to Epstein-Barr virus,herpes simplex type 1, cytomegalovirus and measles virus in psychiatric patients

Summary Distribution of antibodies to herpes simplex type 1 (HSV₁), Epstein-Barr virus (EBV), cytomegalovirus (CMV) and measles virus (MV) was studied in sera and cerebrospinal fluids (CSF) of 41 patients with schizophrenia, 27 patients with primary affective disorders and 25 control patients with neurological diseases.No significant differences in distribution and mean geometric titers (GMT) of antibodies to HSV₁ between the psychiatric and control groups were found. Distribution and GMT of antibodies to EBV were highly significant in psychiatric patients as compared to controls with highest titers in the affective disorder group. Antibodies to HSV₁ were present in 15 CSF specimens of psychiatric patients with reduced CSF/serum ratio in 4, and low levels of antibodies were detected in 8 control patients. Antibodies to EBV-VCA were detected in 4 CSFs of psychiatric patients. Total protein levels were determined in CSF specimens and no correlation with antibodies was found. No significant differences in distribution of antibodies to CMV or MV in the three study groups were found. No antibodies to CMV were demonstrated in CSFs and in one specimen from a patient and two controls antibodies to MV were detected.     

Restriction of latent herpes virus infection in rabbits immunized with subviral herpes simplex virus vaccine

The ability of an experimental herpes simplex virus type 1 (HSV) vaccine to influence the establishment of latent infection was examined. The virion-free vaccine was prepared from HSV 1-infected LEP cells by extraction with Nonidet P 40. Albino rabbits were immunized with three doses of the subviral vaccine (strain KOS) and challenged with 10(5) PFU of HSV 1 (Kupka strain) into the right scarified cornea. Non-immune controls and rabbits immunized with formalin-inactivated virion vaccine (strain KOS) were infected in a similar way. At intervals from 53 to 164 days p. i., the animals were killed and fragments of both trigeminal ganglia were kept separately in culture for 10 days. In 18 out of 22 immunized animals (81.8%), latency was established in the homolateral Gasserian ganglion. The proportion of HSV-yielding fragments, was considerably higher in ganglia from non-immune animals (42.6%) as compared to those from ganglia of rabbits immunized with the subviral vaccine (5.4%) or with the whole virion vaccine (14.6%). The immunity resulting from previous vaccination restricted about 5 times the number of ganglion cells, which become virus carriers.     

Functional cross-reactivity between the glycoprotein B of herpes simplex virus type 1 and Epstein-Barr virus

A monoclonal antibody (T157) directed against gB-1, the glycoprotein B (gB) of herpes simplex virus-1 (HSV-1) shows positive indirect immunoflourescent staining with an Epstein-Barr virus (EBV)-transformed lymphoblastoid cell line B95-8. SDS PAGE and Western blot analysis show that B95-8 cells contain a 110,000 MW protein that co-migrates with the 110,000-115,000 MW gB-1. The gB-1 homologue of EBV (gB-EBV), immunopurified using a T157 affinity column, cross-stimulates HSV-1 immune T cells to proliferate in vitro. Mice immunized by a single subcutaneous injection of 30 microg gB-EBV in saline developed significant protection against HSV-1 challenge infection. Therefore gB-EBV can be considered a potential candidate vaccine and as an antigen to examine the cell-mediated immune response mounted by the host to limit virus spread during productive infection. The significance of a better understanding of the immune response to this and other EBV proteins of productive infection as an alternative to limit tumour growth by preventing virus spread is discussed.     

Efficacy of herpes simplex virus type 1 immunization in protecting against acute and latent infection by herpes simplex virus type 2 in mice.

ICR mice were immunized with herpes simplex virus type 1 (HSV-1) and later challenged with HSV-2 by footpad inoculation. Both immunized animals and age-matched, nonimmunized controls were observed for ascending neurological disease and latent infection of spinal ganglia resulting from the HSV-2 challenge. Control animals had a 78% incidence of acute and latent infection compared with a 1.7% incidence in immunized mice. The data show immunity to HSV-1 is protective against both acute and latent infection by HSV-2.     

Cell-mediated immunity in mice with primary,secondary and latent herpes simplex virus infection

Summary Cell mediated immunity was studied by a cytopathic effect inhibition assay in mice infected in the ear with herpes simplex virus type 1 (HSV 1). Activity appeared rapidly, reaching a high level 6 days after primary infection. It had fallen 10 days after infection and was undetectable during latency, 3–5 weeks after infection. The activity reappeared even more rapidly and strongly after reinoculation with the virus, but stimuli designed to induce recurrent disease did not induce clinical disease in the animals and no activity was detected in them.The activity, which was specific for HSV, was shown to be mediated by T-lymphocytes.With 1 Figure     

巢式PCR检测龈下菌斑中HCMV、EBV、HSV-1与慢性牙周炎活动性的关系

目的　建立临床检测龈下菌斑标本中人巨细胞病毒 (HCMV)、Epstein Barr病毒 (EBV)、单纯疱疹病毒 1型 (HSV 1 )巢式PCR方法 ,研究这 3种病毒与慢性牙周炎活动性的关系。方法　收集6 2例慢性牙周炎患者 (男性 2 7例、女性 35例 ,平均年龄 5 3岁 )的牙周炎活动部位、牙周炎静止部位的龈下菌斑 ,提取DNA后使用巢式PCR检测HCMV、EBV、HSV 1 ,比较分析其在同一病人不同部位的检出率。结果　牙周炎活动部位的HCMV检出率为 38.7%,EBV的检出率为 5 8%,HSV 1的检出率为30 .6 %,两种以上病毒合并感染的检出率为 4 0 .3%;牙周炎静止部位的HCMV检出率为 1 4 .5 %,EBV为 2 2 .6 %,HSV 1为 1 1 .3%,两种以上病毒合并感染的检出率为 1 1 .3%。这 3种病毒及其合并感染在牙周炎活动部位的检出率均高于牙周炎静止部位 ,差异有统计学意义 (P <0 .0 5 )。结论　提示HC MV、EBV、HSV 1与慢性牙周炎的活动性相关。     

Detection of cytomegalovirus, parvovirus B19 and herpes simplex viruses in cases of intrauterine fetal death: association with pathological findings

There are previous indications that transplacental transmission of cytomegalovirus (CMV), parvovirus B19 (PB19) and herpes simplex virus types 1 and 2 (HSV-1/2) cause fetal infections, which may lead to fetal death. In a prospective case-control study we examined the incidence of these viruses in intrauterine fetal death and their association with fetal and placenta pathological findings. Molecular assays were performed on placenta tissue extracts of 62 fetal deaths and 35 controls for the detection of CMV, PB19 and HSV-1/2 genomes. Formalin-fixed, paraffin-embedded liver, spleen and placenta tissues of fetal death cases were evaluated histologically. Thirty-four percent of placental specimens taken from intrauterine fetal deaths were positive for any of the three viruses (16%, 13%, and 5% positive for CMV, PB19, and HSV-1/2, respectively), whereas only 6% of those taken from full term newborns were positive (P = 0.0017). No dual infection was observed. This difference was also observed when fetal deaths with a gestational age <20 weeks or a gestational age >20 weeks were compared with the controls (P = 0.025 and P = 0.0012, respectively). Intrauterine death and the control groups differed in the detection rate of CMV DNA (16% and 3%, respectively; P = 0.047), which was more pronounced in a gestational age >20 weeks (P = 0.03). Examination of the pathological findings among the PCR-positive and PCR-negative fetal deaths revealed that hydrops fetalis and chronic villitis were more common among the former group (P = 0.0003 and P = 0.0005, respectively). In conclusion, an association was detected between viral infection and fetal death, which was more pronounced in the advanced gestational age. Fetal hydrops and chronic villitis were evidently associated with viral DNA detection in cases of intrauterine death.     

Detection of parvovirus B19 capsid proteins in lymphocytic cells in synovial tissue of autoimmune chronic arthritis.

The pathogenic influence of viral agents in chronic inflammatory joint diseases like rheumatoid arthritis has been discussed for many years. More recently, DNA of several viruses, among them parvovirus B19 (B19), was traceable by PCR analysis in synovial fluid and synovial tissue. To investigate the potential role of parvovirus B19 in rheumatoid arthritis, we analyzed the expression of B19 VP1/VP2 proteins by immunohistochemistry in paraffin sections of 63 synovial specimens in rheumatoid arthritis (RA; n = 29), psoriatic arthritis (PSA; n = 6), nonspecific arthritis or synovitis (n = 26), and normal synovia (n = 2). Thereby we could demonstrate replicative virus infection in a variable number of cells in about 90% of rheumatoid specimens and in four of six (66%) cases of psoriatic arthritis, but only in 38% of cases with chronic reactive inflammation and one case of normal synovia. In virus-positive rheumatoid specimens, moreover, the average number of affected cells was significantly higher than in virus-expressing synovia of nonspecific reactive inflammation. These findings support the importance of B19-viral infection in the pathogenesis of chronic arthritis. B19-positive cells in the synovia could be ascribed to CD20- or CD3-positive B- or T-lymphocytes by double immunostaining. Based on these results, B19 infection of lymphocytic cells also seems possible.     

Prevalence of papillomavirus,Epstein-Barr virus,cytomegalovirus, and herpes simplex virus type 2 in urinary bladder cancer

Recent epidemiological studies suggest that the risk for urological malignancies may be related to the exposure to infectious agents. Human Papillomaviruses type 16 and 18 (HPV 16, HPV 18), Epstein-Barr virus (EBV), cytomegalovirus (CMV) and herpes simplex virus type 2 (HSV-2) have been suggested previously as cofactors in the pathogenesis of some malignancies in humans. The present paper, the presence of HPV 16, HPV 18, EBV, CMV and HSV-2 genomes was investigated in a panel of 35 biopsies from urinary bladder carcinomas using the polymerase chain reaction (PCR). Sequences of EBV, HPV, CMV and HSV-2 genomes were detected in 34%, 31%, 11% and 9% of tissue samples respectively, while in 20% of patients we found more than one viral infection. Absence of viral genomes was found in normal bladder. To our knowledge, this is the first report concerning the association of EBV, CMV and HSV-2 with bladder cancer. This finding may raise the question whether such viral infection may contribute to development and progression of some types of urological malignancies in humans. J. Med. Virol. 55:262–267, 1998. © 1998 Wiley-Liss, Inc.     

Herpes simplex virus infection in guinea pigs: an animal model for studying latent and recurrent herpes simplex virus infection.

Footpad infection of guinea pigs with herpes simplex virus led to an acute local inflammatory reaction, followed by a persistent latent infection of lumbosacral dorsal root ganglia. Spontaneous reactivation of the latent virus occurred, leading to recurrent lesions at the site of the initial infection. Clinical observations and virological studies during the acute latent and recurrent infection are reported.     

Latent Epstein-Barr virus (EBV) infection and cytomegalovirus (CMV) infection in synovial tissue of autoimmune chronic arthritis determined by RNA- and DNA-in situ hybridization.

In rheumatoid arthritis (RA) viral triggers, especially Epstein-Barr virus (EBV) and cytomegalovirus (CMV), have been suggested. By PCR analysis DNA of several viruses among which EBV, CMV, and parvovirus B19 (B19) has been detected in RA synovial fluid and synovial tissue. In 63 synovial tissues of 29 rheumatoid arthritis (RA), 6 psoriatic arthritis (PsA), 26 reactive arthritis/synovitis (rA/S), and two normal synovial cases, we recently could demonstrate a high percentage of replicative B19 infection within the synovial tissue, being significantly more frequent in autoimmune arthritis. To further investigate the influence of synovial virus infections in rheumatoid arthritis, we now analyzed the same sample of synovial tissues for CMV and EBV infections by DNA-in situ hybridization (CMV), EBER1/2-RNA-in situ hybridization (EBV), and immunohistochemistry. A significant latent EBV infection of synovial lining cells, synovial fibroblasts, and/or infiltrating lymphocytes was identified in 5/29 (17.2 %) RA, 1/6 (16.7%) PsA, and to a much lower degree in 1/26 (3.8%) rA/S specimens. CMV-DNA was detected in 31% of RA, 3/6 (50%) of PsA, and 11.5% of rA/S. Immunohistochemical analysis of CMV early antigen revealed replicative CMV activity in 20.7% of RA and 2/6 (33.3%) of PsA specimens but not in reactive arthritis synovia. Comparative analysis of the EBV-, CMV-, and published B19-data demonstrated that relevant synovial virus infections in general and furthermore double or multiple infections are far more common in autoimmune arthritis than in rA/S. A triple virus infection was found solely in RA in 10.3% of cases. The evidence of increased synovial persistence of EBV, CMV, or B19 either alone or even more as coinciding infections may further reinforce the notion of a primary role of these viruses in autoimmune arthritis.     

Immunization with a replication-defective herpes simplex virus 2 mutant reduces herpes simplex virus 1 infection and prevents ocular disease

Ocular infections with herpes simplex virus 1 can lead to corneal scarring and blindness, with herpes keratitis being the major infectious cause of blindness. There is currently no clinically approved vaccine and nearly all developmental vaccines are targeted against HSV-2 and genital herpes. We tested the ability of an HSV-2 replication-defective virus, a genital herpes vaccine candidate, to protect against HSV-1 corneal infection. Immunization with HSV-2 dl5-29 reduced viral replication in the cornea, prevented ocular disease and reduced latent infection by the HSV-1 strain. Therefore, this HSV-2 replication-defective mutant strain may have applications for prevention of herpes keratitis and genital herpes due to HSV-1 infection.     

Acute and latent infection of sensory ganglia with herpes simplex virus: immune control and virus reactivation.

The role of antiviral antibody in controlling the acute and latent phases of herpes simplex virus (HSV) infection in sensory ganglia of mice was studied in vitro and in vivo. Organ cultures of ganglia inoculated in vitro with HSV produced infectious virus for at least 3 weeks. In the presence of antiviral antibody, the titre of virus was markedly reduced, but the infection was not eliminated. Similarly, passive administration of antibody to HSV-infected immunodeficient (nude) mice reduced the virus titre but did not eliminate the acute phase of the ganglionic infection. Suppression of the cell-mediated immune response in latently infected immunocompetent mice by treatment with cyclophosphamide and/or X-irradiation resulted in reactivation of HSV in up to 70% of the animals. Reactivation was demonstrated by recovering infectious virus in cell-free homogenates of ganglia and eye globes and by finding virus antigens in ganglia by immunofluorescent staining. Reactivation occurred both in vitro and in vivo in the presence of high concentrations of neutralizing antibody. It is concluded that antibody alone is not sufficient to eliminate the acute phase of the ganglionic infection and that cytotoxic agents known to suppress the host's cellular immune response can reactivate virus in the presence of neutralizing antibody.     

Virus infection in patients with histiocytic necrotizing lymphadenitis in Taiwan. Detection of Epstein-Barr virus, type I human T-cell lymphotropic virus, and parvovirus B19

The relationship of Epstein-Barr virus (EBV), type I human T-cell lymphotropic virus (HTLV-I), and parvovirus B19 to histiocytic necrotizing lymphadenitis was studied prospectively in 10 Taiwanese patients using materials obtained by fine-needle aspiration and lymph node biopsy. The presence of EBV was detected by in situ hybridization for EBV-encoded RNA expression. Immunocytochemistry was used to detect virus-encoded protein for EBV and parvovirus B19. DNA in situ hybridization and polymerase chain reaction were performed to determine the existence of HTLV-I provirus. Expressions of EBV-encoded RNA and Fas ligand were detected in all cases. Expression of EBV-encoded protein was identified in only 1 case. Neither HTLV-I nor parvovirus B19 was detected in any case.     

Immune response and latent infection after topical treatment of herpes simplex virus infection in hairless mice.

Treatment of herpes simplex virus (HSV)-infected hairless mice with a 2% phosphonoacetic acid (PAA) ointment prevented the appearance of virus-induced skin lesions and subsequent central nervous system (CNS) involvement. Treatment started 24 h after infection significantly reduced the intensity of the skin lesions and also prevented CNS involvement. After four to six applications of PAA ointment, a moderate skin erythemia developed, followed by scaling and complete healing 7 days after cessation of the treatment. Mice treated early after HSV infection had low or undetectable levels of virus-specific antibodies but were completely resistant to reinfection. Early treatment prevented the development of a latent ganglionic infection, but treatment initiated 24 h after infection could not prevent the establishment of the latent infection. PAA-treated and HSV-infected mice with nondetectable levels of antibodies did not develop, with a single exception, a latent ganglionic infection unpon reinfection. The cell-mediated immune response determined by levels of [14C]thymidine incorporation in Ficoll-Hypaque-purified spleen lymphocytes cultures was low in PAA-treated mice; it increased slightly after challenge infection but was strong in mice that proved to harbor a latent HSV infection in the ganglia.