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PubMedDate not supplied

[Pathogenesis, diagnosis and therapy of heart disease caused by enteroviruses (molecular biological study)].

Molecular hybridization studies have demonstrated that human enteroviruses, including group B coxsackieviruses (CVB), are detectable in myocardial tissue of patients with acute and chronic myocarditis. As well, such infections are observed in some patients with end-stage dilated cardiomyopathy indicating the possibility of persistent heart muscle infection. Enterovirus persistence in the human heart is supported by the discovery in various murine models of chronic myocarditis, demonstrating that coxsackievirus B3 (CVB3), typically a cytolytic virus, is capable of evading immunological surveillance in a host-dependent manner. Currently attention is focused on the analysis of molecular mechanisms of virus persistence, the characterization of viral and host factors and their impact in determining the natural course of myocardial enterovirus infections. The evidence for a causal linkage of enterovirus infection with heart muscle diseases has emerged therapeutic implications. From the view of a virologist, immunosuppressive treatment of patients revealing enterovirus infection in the myocardium with steroids is clearly contraindicated. The evaluation of potent antiviral agents, such as interferons, in established in vitro and in vivo model systems of enterovirus infection is expected to contribute significantly to new therapeutic strategies in human enteroviral heart disease.

Animals↗
PubMedDate not supplied

Neurologic involvement in an outbreak of enterovirus 71 infection: a hospital-based study.

Enterovirus (EV) can cause varied clinical manifestations. Involvement of the central nervous system (CNS) with the nonpolio EVs are common and important causes of morbidity in children. To investigate the manifestations of nonpolio enteroviral infections with CNS involvement during the EV outbreak, from February 1998 to January 1999, we collected 153 hospitalized patients in our pediatric ward caused by nonpolio EV infections which were diagnosed by history, clinical features, or detected from viral cultures. Fourteen patients (9.2%) had CNS presentations, 13 males and one female. The ages ranged from one month to 10.3 years. The spectrum of CNS presentations included aseptic meningitis (4 cases, 28.6%), encephalitis (5 case, 35.7%), encephalomyelitis (3 cases, 21.4%), and poliomyelitis-like syndrome (2 cases, 14.3%). Among these patients, 8 cases (57.1%) were isolated with EV71 from at least one site of rectal or throat swab sampling. Two fatal cases were presented as encephalitis and complicated with pulmonary edema. Generally, enteroviral infections are considered as a benign infectious disease in children. However, pediatricians should keep in mind that EV71 has caused several endemic outbreaks and continues to be an occasional cause of severe CNS disease. Early evaluation and appropriate treatment of CNS enteroviral infections may minimize the neurologic sequelae.

Central Nervous System Viral Diseases↗
PubMed2003

Cardiac troponin I: a reliable marker and early myocardial involvement with meningoencephalitis after fatal enterovirus-71 infection.

OBJECTIVES: A major outbreak of enterovirus 71 (EV71) in Taiwan in 1998 caused many severe cases and 78 deaths. Our purpose was to find reliable markers and early indicators of fatal EV71 central nervous system (CNS) infection. METHODS: From June 2000 to November 2001, 21 patients with hand foot mouth disease or herpangina with CNS infection were admitted to Kaohsiung Veterans General Hospital. All 21 had culture-confirmed EV71 infection or were EV71 IgM positive. Patients were divided into two groups: group I included the five fatalities at our institution and group II, the 16 surviving patients. RESULTS: Of the 21 infants and children with EV71 infection with CNS involvement, MR imaging studies were completed on 17, and 15 showed hyperintensity in the posterior portions of brain stem. All patients received intravenous immunoglobulin (IVIG) 1 g/day for two days and supportive care. Five patients rapidly deteriorated owing to irreversible hypotension and died. The other 16 patients recovered completely without sequel. In group I patients, the decrease of cardiac ejection function is significant and laboratory findings showed lower platelet count (P=0.0192). The mean of initial cTnI level for groups I and II was 10.6+/-11.6 and 0.48+/-0.55 ng/dl, respectively, higher in group I than in II (P=0.0019). CONCLUSION: We hypothesized that like patients with severe burns, those with severe EV-71 CNS meningoencephalitis have varying degrees of non-ischemic cardiac injury, manifesting as leakage of cTnI from myocytes into the circulation. EV-71 CNS meningoencephalitis likely to die with an early myocardial involvement evidenced by reduced ejection fraction and release of cTnI. We conclude that fatal EV71 CNS infection quickly leads to death due to severe encephalopathy associated with cardiomyopathy.

Biomarkers↗
PubMed2003

Enterovirus 71 infection induces apoptosis in Vero cells.

The effects of Enterovirus 71 (HEV71) infection on African green monkey kidney cells (Vero) were investigated. It was found that the infected cells showed progressive cellular morphological changes characteristic in apoptotic cells within 10 hours post-infection. The number of apoptotic cells correlated significantly with the number of HEV71 antigen positive cells when cells were labeled using terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) and stained for HEV71 antigen. Approximately 11, 26, 45 and 50% of the infected cells were apoptotic at 12, 24, 48 and 72 hours post-infection, respectively. Internucleosomal DNA fragmentation, characteristic in the late stage of apoptosis was noted beginning on day 2 post-infection. The DNA fragmentation, however, was absent in cells treated with the heat- and ultraviolet light-inactivated virus inocula. These results demonstrate the capacity of HEV71 to induce apoptosis in the infected cells. The induction, however, requires high level of HEV71 infectivity and the presence of live virus particles, suggesting the need for the presence of specific viral proteins for apoptosis to occur.

Animals↗
PubMed2000

Congenital enterovirus 71 infection: a case study with virology and immunohistochemistry.

Previously reported enterovirus 71 (EV71) infections associated with hand-foot-mouth disease, aseptic meningitis, encephalitis, polio-like myelitis, and paralysis all have involved young children. We report a 28-year-old woman who possibly contracted EV71 infection during pregnancy. Obstetric ultrasonograms at 25 weeks of gestation revealed an abnormal placenta, as well as hepatosplenomegaly, liver calcification, excessive ascites, and mild hydrocephalus of the fetus. Presence of EV71 was determined by immunodot blotting, virus culture, and partial sequencing of the amplified product of nested reverse transcription-polymerase chain reaction. Postmortem immunohistochemistry further identified EV71 in the fetal midbrain and liver. The findings indicate that intrauterine EV71 infection is possible during pregnancy.

Adult↗
PubMed2001

Acute flaccid paralysis in infants and young children with enterovirus 71 infection: MR imaging findings and clinical correlates.

BACKGROUND AND PURPOSE: Enterovirus 71 (EV71) infection is now considered an important cause of childhood acute flaccid paralysis. The purpose of our study was to determine whether EV71-infection-related acute flaccid paralysis in infants and young children has characteristic MR imaging patterns. METHODS: Seven infants and young children with acute paralysis of the upper or lower extremities and positive EV71 cultures underwent spinal MR studies during an outbreak of hand-foot-and-mouth disease in Taiwan in 1998. RESULTS: Acute paralysis was observed in one upper extremity in two patients, in one lower extremity in three patients, and in both lower extremities in two patients. None of the patients had sensory impairment or bulbar palsy. MR studies showed unilateral or bilateral hyperintense lesions in the anterior horn regions of the cord on T2-weighted images in six patients. No abnormal signal was present in one patient. Two of three patients who received intravenous injections of contrast material had ventral root enhancement on T1-weighted images. One of them also had enhancement of the unilateral anterior horn cells. At clinical follow-up, both patients with bilateral anterior horn abnormalities had residual motor weakness, whereas only one of the five patients with unilateral involvement had residual weakness. CONCLUSION: EV71 radiculomyelitis tends to be unilateral and to specifically involve both the anterior horn cells of the cord and the ventral roots. MR imaging allows early detection of spinal cord and root lesions.

Acute Disease↗
PubMed2006

Protection of neonatal mice from lethal enterovirus 71 infection by maternal immunization with attenuated Salmonella enterica serovar Typhimurium expressing VP1 of enterovirus 71.

This study describes the potential use of attenuated Salmonella enterica serovar Typhimurium strains to express and deliver VP1 of enterovirus 71 (EV71) as a vaccination strategy to prevent EV71 infection in mice. When orally administered to BALB/c mice, both attenuated carrier strains, CNP101 and SL7207, were able to efficiently invade livers and spleens, while only the virulence plasmid-carrying strain SL7207 persisted for more than 30 days in these organs. A recombinant in vivo-regulated promoter expression plasmid expressing VP1 antigen of EV71 was constructed. The expression of the VP1, directed by the pagC promoter, in attenuated Salmonella was confirmed by Western blot hybridization. Both humoral and cellular immune responses were elicited in mice by oral immunization with such Salmonella-based VP1 vaccines. We evaluated the protective efficacy of the vaccines in mice using a maternal immunization protocol. With a lethal challenge, ICR newborn mice born to dams immunized with Salmonella-based VP1 vaccine showed a 50-60% survival; in contrast, none of the mice in the control group survived the challenge. Our data indicated that Salmonella-based VP1 subunit vaccines are a promising vaccine strategy in the prevention of EV71 infection.

Animals↗
PubMed1999

[Efficacy of immunized milk for preventing viral infection].

This paper investigated the efficacy of passive protection provided by milk (immunized milk) against enterovirus infection in mice experimentally infected with enterovirus. Milk with a high antibody titer against six enterovirus serotypes was prepared from hyperimmunized goat. In vivo and in vitro experiments were performed and the results showed that immunized milk has an antiviral activity against enterovirus infection. Further observation was performed using Coxsackie B 3 virus (CVB 3). When immunized milk was orally applied to mice prior to oral inoculation with CVB 3, preventive effects against viral infection such as reduction of histopathological changes in the heart and reduced detection of the virus genome in the organs were seen. The antiviral effect was also indicated by the increase of CD4+T cells proportion in the i-IEL. The proportion of virus specific CD4+T cells was increased in mice treated with immunized milk, whereas no such increase was detected in control mice. These results suggest that oral application of immunized milk is not only capable of preventing viral infection but also induces specific immunological responses. These phenomena may play an important role in host defense mechanisms.

Animals↗
PubMed1988

Enterovirus in pregnant women and the perfused placenta.

A survey of all nonpolio enterovirus infections in adults for a 10-year period revealed that most such infections will occur in the child-bearing age group during the third quarter of the year. Coxsackieviruses occurred more often than echovirus infection. Fourteen pregnant women, including four with meningitis, who had an enterovirus infection from 16 to 37 weeks of gestation delivered uninfected, healthy infants. Attempts to demonstrate transplacental passage of Coxsackie B-3 or ECHO-11 viruses by use of the dual, recirculating, in vitro perfusion of an isolated placental lobule were unsuccessful even with a maternal virus input of 200,000 median tissue culture infectious doses. This suggests that transplacental passage of virus does not occur readily, and that most neonates of infected mothers will be unharmed.

Adolescent↗
PubMed2006

Transcriptomic and proteomic analyses of rhabdomyosarcoma cells reveal differential cellular gene expression in response to enterovirus 71 infection.

Insights into the host antiviral strategies as well as viral disease manifestations can be achieved through the elucidation of host- and virus-mediated transcriptional responses. An oligo-based microarray was employed to analyse mRNAs from rhabdomyosarcoma cells infected with the MS/7423/87 strain of enterovirus 71 (EV71) at 20 h post infection. Using Acuity software and LOWESS normalization, 152 genes were found to be downregulated while 39 were upregulated by greater than twofold. Altered transcripts include those encoding components of cytoskeleton, protein translation and modification; cellular transport proteins; protein degradation mediators; cell death mediators; mitochondrial-related and metabolism proteins; cellular receptors and signal transducers. Changes in expression profiles of 15 representative genes were authenticated by real-time reverse transcription polymerase chain reaction (RT-PCR), which also compared the transcriptional responses of cells infected with EV71 strain 5865/Sin/000009 isolated from a fatal case during the Singapore outbreak in 2000. Western blot analyses of APOB, CLU, DCAMKL1 and ODC1 proteins correlated protein and transcript levels. Two-dimensional proteomic maps highlighted differences in expression of cellular proteins (CCT5, CFL1, ENO1, HSPB1, PSMA2 and STMN1) following EV71 infection. Expression of several apoptosis-associated genes was modified, coinciding with apoptosis attenuation observed in poliovirus infection. Interestingly, doublecortin and CaM kinase-like 1 (DCAMKL1) involved in brain development, was highly expressed during infection. Thus, microarray, real-time RT-PCR and proteomic analyses can elucidate the global view of the numerous and complex cellular responses that contribute towards EV71 pathogenesis.

Cell Line, Tumor↗
PubMed2002

Left ventricular dysfunction in children with fulminant enterovirus 71 infection: an evaluation of the clinical course.

We describe 2 children with typical hand, foot, and mouth disease due to enterovirus 71 infection, 1 of whom died. Both cases were complicated by acute fulminant shock syndrome; the patients had remarkable acute left ventricular dysfunction. The clinical experience indicates that the rapid death associated with fulminant enterovirus rhombencephalitis is the result of rapid cardiogenic shock rather than neurogenic pulmonary edema.

Child, Preschool↗
PubMed2004

Outcome of enterovirus 71 infections with or without stage-based management: 1998 to 2002.

BACKGROUND: Enterovirus 71 (EV71) infection may progress through four stages, one of which is cardiopulmonary failure. In Taiwan in 1998 almost all the EV71 patients with cardiopulmonary failure died. To improve clinical outcome of EV71 patients, we developed a stage-based management program in 2000. METHODS: The medical records of 196 EV71 patients who did not have stage-based management (1998 to 1999) and of 331 EV71 patients who did (2000 to 2002) at Chang Gung Children's Hospital were reviewed for demographic characteristics, clinical syndromes, case-fatality rates and sequelae. We compared and analyzed the results for the 2 groups. RESULTS: Of the patients who did not receive stage-based management, 83% (15 of 18) of cases with both central nervous system (CNS) involvement and cardiopulmonary failure died during the acute stage of the infection. Two patients died at convalescence, and 1 had sequelae of dysphagia and limb weakness. By contrast of the patients who received stage-based management, 33% (12 of 36) of patients with CNS and cardiopulmonary failure died during the acute stage, 8% (3 of 36) died at convalescence, 14% (5 of 36) recovered and 43% (16 of 36) had severe sequelae of central hypoventilation, dysphagia and limb weakness (P < 0.001). For cases with CNS and cardiopulmonary failure, multivariate analysis showed that age older than 2 years and cerebro-spinal fluid white blood cell count >100/microl were associated with a increase in acute mortality [95% confidence interval (CI) 1.9 to 105.3, P = 0.001; 95% CI 1.1 to 66.6, P = 0.04, respectively], but stage-based management was significantly associated with a reduction in acute mortality (95% CI 0.007 to 0.24; P = 0.0004). Stage-based management did not affect the outcome of cases with CNS involvement alone. CONCLUSIONS: Stage-based management reduced the case fatality rate of EV71-related cardiopulmonary failure, but two-thirds of the survivors had severe sequelae.

Case-Control Studies↗
PubMed2005

Enteroviruses: new findings on the role of enteroviruses in type 1 diabetes.

Common enterovirus infections appear to initiate or facilitate the pathogenetic processes leading to type 1 diabetes, and sometimes also precipitate the clinical disease. It is not known in detail how enterovirus infections bring about the loss of insulin-producing beta-cells, a phenomenon characteristic of the disease. Recent results from studies on pancreases from human autopsies and cultured human islets support the idea that during systemic enterovirus infections, the virus may reach pancreatic islets and cause direct beta-cell damage. Although individual enteroviruses (EV) exhibited differences in their beta-cell tropism in the cultured human islets, all serotypes studied contained highly destructive strains. The final confirmation on the role of enteroviruses in type 1 diabetes can only be obtained from intervention studies. If the association holds true then it would be possible to reduce the risk of developing type 1 diabetes by preventing enterovirus infections with a multivalent enterovirus vaccine that could be given to children soon after birth.

Diabetes Mellitus, Type 1↗
PubMed2002

Differential display RT-PCR analysis of enterovirus-71-infected rhabdomyosarcoma cells reveals mRNA expression responses of multiple human genes with known and novel functions.

In order to better understand cellular responses to viral infection at the transcriptional level, we employed differential display RT-PCR to analyze mRNAs from RD rhabdomyosarcoma cells following infection with a neurovirulent enterovirus 71 (EV71) strain, compared with mRNAs from uninfected cells. Of 250 expressed sequence tags (ESTs) isolated, sequenced, and identified, all were of cellular origin except 1 that was of viral origin. Of these, 156 were individual distinctive clones, comprising 45 mRNAs showing unaltered expression and 111 mRNAs exhibiting upregulation or downregulation. Of the 45 uniformly expressed mRNAs, 14 represented unknown genes. Of the 111 differentially expressed mRNAs, 63 did not match any known genes. Forty-eight of the 111 mRNAs modified by EV71 infection matched known genes, including those encoding components of cell cycle, cytoskeleton, and cell death mediators; protein degradation mediators; mitochondrial-related proteins; components of protein translation and modification; and cellular transport proteins. The altered expression profiles of representative genes were authenticated by semiquantitative RT-PCR and real-time RT-PCR. We also identified a novel alternatively spliced transcript of TRIP7 thyroid receptor interactor protein; the putative human homolog of murine mc7 mRNA predominantly expressed in the brain; and a novel mRNA similar to that encoding vacuolar protein 8 involved in protein targeting. These results underscore the applicability of the mRNA differential display technique for elucidating the expression profiles of known and even novel genes in response to cellular infection with pathogenic viruses.

Amino Acid Sequence↗
PubMed1993

Molecular biology of viral heart disease.

In situ hybridization studies have proved that myocardial enterovirus infections are detectable in all stages of acute and chronic enterovirus-induced myocarditis as well as in some patients with end-stage dilated cardiomyopathy, suggesting the possibility of myocardial enterovirus persistence. Possible enterovirus persistence in the human heart is supported by the discovery of enterovirus persistence in different murine models of chronic myocarditis, demonstrating that coxsackie-virus B3, typically a cytolytic enterovirus, is capable of evading immunological surveillance in a host-dependent fashion. Progress is currently being made in unraveling the molecular mechanisms of enterovirus persistence, the diversity of host and virus genetics and their impact on the nature and severity of the disease. Apart from providing an etiologic diagnosis, there are therapeutic implications from in situ demonstration of myocardial enterovirus infection. Evaluation of specific antiviral agents, for example interferons, may lead to the development of new therapeutic strategies capable of providing protection against myocardial enterovirus infection.

Cardiomyopathy, Dilated↗
PubMed2002

Responses of IgM for enterovirus 71 infection.

A rapid serological assay was developed for detection of specific IgM to enterovirus 71, a human picornavirus that is usually associated with severe central nervous system complications. The sensitivity and specificity of this "in-house" mu-capture enzyme linked-immunosorbent assay was assessed by testing 213 serum samples. With the conventional virus culture as a standard method, the sensitivity and specificity were 91.5 and 93.1%, respectively, for this newly developed immunoassay. This method allows for detection of the IgM responses from the patients either infected by genotype B or genotype C of enterovirus 71. IgM can be detected as early as the second day from the onset of disease. IgM responses exhibit 100% positive rate from enterovirus 71-infected patients with complications, including encephalitis, meningitis, polio-like syndrome, pulmonary edema, and fatal cases. These findings suggest that detection of specific IgM by the use of enzyme linked-immunosorbent assay is a rapid and valuable way for the diagnosis of enterovirus 71 infection.

Antibodies, Viral↗
PubMedDate not supplied

[Superacute lethal liver necrosis in monkeys infected with highly pathogenic variants of enteroviruses (ECHO 11 and ECHO 19 viruses)].

The authors studied the phenomenon of superhigh virulence for monkeys of viruses ECHO 11 (E11) and ECHO 19 (E19) isolated in 1981-1991 in Russia and Tadzhikistan from children with acute enterovirus uveitis or severe enterovirus infection. In 21 morphologically examined animals after coma, macrofocal or total necrosis of hepatocytes in weak inflammation was seen as early as experiment day 1-6. Hepatic lesion interpreted as hepatosis underlies acute hepatic failure with lethal outcome. The most intensive reproduction of viruses E11 and E19 occurred in the liver. Changes in the kidneys, lungs, spleen, adrenals, CNS detected in many animals aggravated the disease, but were not the primary cause of death. The disease in monkey was similar by the main criteria to superacute lethal diseases (acute hepatic failure against massive hepatic necrosis, hemorrhagic syndrome) registered in outbreaks of E11 and E19-virus infection in children in the USA, Great Britain, Israel, Russia and Ukraine. Monkeys can be used as man-adequate model for study of enterovirus superacute lethal necrosis of the liver.

Animals↗