Chimeric virus-like particles presenting common neutralizing epitopes of enterovirus 71
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To develop an effective vaccine against multiple genotypes of enterovirus 71 (EV71), we generated chimeric virus-like particles (VLPs) repetitively displaying the common neutralizing epitopes of EV71 and evaluated their immunogenicities in mice.The two conserved epitopes, encompassing amino acids 163-177 and 208-222 of VP1 of EV71, were fused to hepatitis B core antigen (HBcAg) and expressed in E.coli.The resulting fusion proteins were found to assemble into chimeric VLPs.Both unmodified HBcAg and chimeric VLPs induced HBcAg-specific antibody responses in mice, however, only chimeric VLP-immunized sera possessed EV71 epitope-specific IgG antibodies and efficiently neutralized different EV71 strains.Collectively, our results indicate that the chimeric VLP is capable of eliciting broadly neutralizing antibody responses and is therefore a promising EV71 vaccine candidate.Keywords:
Enterovirus 71
To study the clinical and epidemiologic characteristics and the serum types of enterovirus of hand, foot and mouth disease (HFMD) in children.The RT-nPCR method was established with universal primers within 5' untranslated region of enterovirus and VP1 region of Coxsackievirus A16 (CAV16) and enterovirus 71 (EV 71). Enteroviruses were detected with RT-nPCR in 237 children with HFMD. Clinical and epidemiologic characteristics and serum types of enterovirus of the patients with HFMD were studied.The patients'age ranged from 7 months to 11 years (mean 4.2 +/- 0.5 years). The majority (94.5%) were less than 6 years old. HFMD was mostly seen in spring and winter (67.9%). Oral mucosal pox or ulcer as well as hand and foot rashes were observed in all 237 patients. Fever occurred in 141 patients (59.5%). Of the 237 patients, 133 (56.1%) were RT-nPCR positive. Of the 133 cases, 38 were positive for EV71, 64 were positive for CAV16, and 31 were negative for both EV71 and CAV16. The patients infected by different types of enteroviruses had similar clinical characteristics. Gene colon and sequence analysis for 12 strains of enteroviruses PCR positive products presented as EV71 (n=5), CAV16 (n=5), ECHO13 (n=1), and CAV5 (n=1).HFMD tends to occur in younger children less than 6 years old. The majority are affected in spring and winter. EV71 and CAV16 are common pathogens of HFMD. There is no relationship between clinical characteristics and serum types of enteroviruses in HFMD patients.
Enterovirus 71
Hand-foot-and-mouth disease
Foot (prosody)
Coxsackievirus
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Poxviridae
Medical microbiology
Veterinary virology
Orthopoxvirus
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Objective To investigate the pathogen distribution of patients with hand, foot and mouth diseases (HFMD) hospitalized in division of pediatrics, Beijing Ditan Hospital, Capital Medical University, in 2010. Methods Real-time fluorescence quantitative RT-PCR kits with three reagents: universal enterovirus primer, Coxsackievirus A16 (CA16) primer and enterovirus 71 (EV71) primer were used to detect the samples. Results The enterovirus-positive rate was 63.20% (213/337).Among this, the positive rates of enterovirus 71, Coxsackievirus A-16 and other types of enterovirus were 40.36% (136/337), 5.46%(19/337) and 17.21%(58/337), respectively. Conclusions In 2010, the etiology of children with hand, foot and mouth disease hospitalized in our hospital was dominated by enterovirus 71(EV71). The infection rates of enterovirus except enterovirus 71 and Coxsackievirus A16 are main pathogens. Non-CA16, non-EV71 virus infection were significant higher than Coxsackievirus A16 infection.
Enterovirus 71
Coxsackievirus
Hand-foot-and-mouth disease
Etiology
Primer (cosmetics)
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The recent, ongoing epidemic of hand, foot, and mouth disease (HFMD), which is caused by enterovirus infection, has affected millions of children and resulted in thousands of deaths in China. Enterovirus 71 (EV71) and coxsackie A16 (CA16) are the two major distinct pathogens for HFMD. However, EV71 is more commonly associated with neurologic complications and even fatalities. Therefore, simultaneously detecting and differentiating EV71 and CA16 specifically from other enteroviruses for diagnosing HFMD is important. Here, we developed a one-step, triplex, real-time RT-PCR assay for the simultaneous detection of EV71, CA16, and pan-enterovirus (EVs) in a single tube with an internal amplification control. The detection results for the serially diluted viruses indicate that the lower limit of detection for this assay is 0.001-0.04 TCID50/ml, 0.02 TCID50/ml, and 0.001 TCID50/ml for EVs, EV71, and CA16, respectively. After evaluating known HFMD virus stocks of 17 strains of 16 different serotypes, this assay showed a favorable detection spectrum and no obvious cross-reactivity. The results for 141 clinical throat swabs from HFMD-suspected patients demonstrated sensitivities of 98.4%, 98.7%, and 100% for EVs, EV71, and CA16, respectively, and 100% specificity for each virus. The application of this one-step, triplex, real-time RT-PCR assay in clinical units will contribute to HFMD surveillance and help to identify causative pathogen in patients with suspected HFMD.
Enterovirus 71
Throat
Hand-foot-and-mouth disease
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Enterovirus 71
Coxsackievirus
Hand-foot-and-mouth disease
Medical microbiology
Enterovirus Infections
Isolation
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Epidemiological data from active surveillance on human enterovirus, which could cause hand, foot, and mouth disease, were limited. An active surveillance system was used to investigate the enterovirus spectrum and the incidence of different enteroviruses in infants aged 6-35 months in Jiangsu Province from 2012 to 2013. Fifty-nine infants were randomly selected from 522 non-EV-A71/CV-A16 HFMD patients. We collected 173 throat swabs and 174 rectal swabs from these infants. RT-PCR was used to amplify 5'-UTR and VP1 regions of enteroviruses and the serotypes were determined by the sequence comparison using BLAST. Twenty-one non-EV-A71/CA16 enterovirus serotypes were detected in those infants. E16, E18 were firstly reported in HFMD patients. The four top common non-EV-A71/CV-A enteroviruses among infants were CV-B3, CV-A10, CV-A6, and E9 with the HFMD incidence rates at 1.4%, 0.84%, 0.56%, and 0.47%, respectively. Over 20.8% patients were co-infected with multiple enteroviruses. Neither the course of sickness nor clinical symptoms of the co-infected patients was more severe than those infected with single enterovirus. Two patients were infected different enterovirus successively within 2 months. Several new enterovirus serotypes and multiple models of infection associated with HFMD were discovered through the active surveillance system. These data provide a better understanding of the viral etiology of HFMD.
Enterovirus 71
Etiology
Throat
Picornavirus
Rhinovirus
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Objectives To understand the difference of clinical features of hand-foot-mouth disease(HFMD) caused by enterovirus 71 and other enterovirus types.Methods A total of 106 children with HFMD were included and divided into three groups according to etiology: group 1(enterovirus 71 infection,n = 63),group 2(Coxsackievirus A16 infection,n = 16) and group 3(other enterovirus types infection,n = 27).The information of fever,rash and clinical manifestations of nervous system,respiratory and circulatory systems,and the laboratory data were summarized and analyzed among the three groups.Results The patients in group 1 had higher incidence of central nervous system infection than that of group 2 and group 3.The patients in group 1 had higher peak temperature,longer fever time and more frequent limb tremor and irritation than that in group 2(P 0.05).The patients of CA16 had higher incidence of headache,vomiting,convulsion than that in group 1(P 0.05).Conclusions Patients with EV71 infection had higher peak temperature,longer fever time,higher incidence of central nervous system infection,more frequent limb tremor and irritation than other enterovirus types infection.These characteristics may help pediatricians to identify the type of enterovirus.
Enterovirus 71
Etiology
Coxsackievirus
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Abstract The hemagglutinin (HA) protein of influenza virus mediates essential viral functions including the binding to host receptor and virus entry. It also has the antigenic sites required for virus neutralization by host antibodies. Here, we characterized an H3N2 triple reassortant (TR) influenza virus (A/turkey/Ohio/313053/04) with a mutation at the receptor binding domain (Asp190Ala) that occurred upon virus transmission from turkeys to pigs in an experimental infection study. The mutant virus replicated less efficiently than the parental virus in human, pig and turkey primary tracheal/bronchial epithelial cells, with more than 3-log 10 difference in virus titer at 72 hours post infection. In addition, the mutant virus demonstrated lower binding efficiency to plasma membrane preparations from all three cell types compared to the parental virus. Antisera raised against the parental virus reacted equally to both homologous and heterlogous viruses, however, antisera raised against the mutant virus showed 4-8 folds lower reactivity to the parental virus.
H5N1 genetic structure
Antibody-dependent enhancement
Veterinary virology
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Few studies have investigated the viral isolation characteristics for severe complicated enterovirus infection (SCEI). This study evaluated the seasonality and contribution of circulated viruses to the chronologic trend and weekly reported SCEI epidemic.Enterovirus infection surveillance and virology laboratory data in 2000 to 2008 obtained from the Centers for Disease Control in Taiwan were analyzed. We measured the monthly and weekly virology isolation rates by viral types. The virus-specific and the season-specific relative risks for SCEI and 95% confidence intervals (CI) associated with the isolated circulating viruses and weather status was evaluated.Among 1539 virology confirmed SCEI cases, the mean annual incidence rates ranged from 0.72/100,000 to 32.5/100,000 in children aged 5 years and less; rates were higher in warm months with cases peaking in June (12.6%). The untypeable nonpolio enterovirus was the most frequently isolated type among the monitored specimens (6.07%), followed by coxsackievirus A (3.99%), EV71 (1.77%), coxsackievirus B (1.56%), and echovirus (1.23%). However, these SCEI cases had very strong associations with the isolation of EV71 and coxsackievirus A and B. The corresponding relative risks were 1.14 (95% CI, 1.11-1.17), 1.03 (95% CI, 1.01-1.04), and 1.09 (95% CI, 1.07-1.12), respectively, for 1% increase in weekly isolation rate.Isolation rates for EV71 and coxsackieviruses A and B can predict the development of SCEI cases, particularly in warm months.
Enterovirus 71
Enterovirus Infections
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SummarySpider monkeys and chimpanzees were given a series of three injections consisting of 17D yellow fever virus, followed by living West Nile virus, followed by a third injection which consisted of formalin-inactivated Russian spring-summer virus vaccine. On the basis of neutralizing antibody responses, the limitation of viremia, or both, developing when the animals were challenged with virulent viruses, these primates were judged to be protected to a considerable extent against Japanese B encephalitis, West Nile virus, St. Louis encephalitis, Murray Valley encephalitis virus, dengue types 1, 2, 3, and 4, two antigenic types of the Russian spring-summer virus complex, and Wesselsbron virus.An isolate of West Nile virus was passed a number of times in chick embryo tissue cultures and purified by the plaque technique. The progeny of two virus plaques, in a concentration of 106 mouse intracerebral lethal doses, did not produce encephalitis in intracerebrally inoculated rhesus monkeys. These attenuated viral preparations, on the basis of intracerebral titrations in mice, had at least 1,000 times the virus concentration that was necessary to produce encephalitis with the parent type. One of these attenuated isolates still produced homologous and heterologous neutralizing antibodies comparable to those of the parent strain. The data indicate that this attenuated West Nile virus did not revert to a more virulent form after alternate intracerebral passages in rhesus monkeys and suckling mice.The TP-21 strain of the Russian spring-summer virus complex was passed a number of times in chick embryo tissue cultures and purified by the plaque technique. The progeny from one of the virus plaques, in a concentration of approximately 300,000 mouse i.c. LD50, did not produce encephalitis when inoculated intracerebrally into rhesus monkeys. When this purified virus isolate of TP-21 was substituted for the formalin-inactivated Russian spring-summer vaccine in the triple vaccination procedure, considerable protection was noted in spider monkeys challenged with four members of the Russian spring-summer group of viruses.
Viremia
Flavivirus
Attenuated vaccine
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