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1.
PLoS Negl Trop Dis ; 18(5): e0012159, 2024 May.
Article in English | MEDLINE | ID: mdl-38739673

ABSTRACT

BACKGROUND: Rodents are recognized as the hosts of many vector-borne bacteria and protozoan parasites and play an important role in their transmission and maintenance. Intensive studies have focused on their infections in vectors, especially in ticks, however, vector-borne bacterial and protozoan infections in rodents are poorly understood although human cases presenting with fever may due to their infection have been found. METHODS: From May to October 2019, 192 wild rodents were trapped in wild environment of Guangxi Province, and the spleen samples were collected to reveal the presence of vector-borne bacterial and protozoan infections in them. The microorganisms in rodents were identified by detecting their DNA using (semi-)nested PCR. All the PCR products of the expected size were subjected to sequencing, and then analyzed by BLASTn. Furthermore, all the recovered sequences were subjected to nucleotide identity and phylogenetic analyses. RESULTS: As a result, 192 rodents representing seven species were captured, and Bandicota indica were the dominant species, followed by Rattus andamanensis. Based on the (semi-)nested PCR, our results suggested that Anaplasma bovis, Anaplasma capra, Anaplasma ovis, Anaplasma phagocytophilum, "Candidatus Neoehrlichia mikurensis", "Candidatus E. hainanensis", "Candidatus E. zunyiensis", three uncultured Ehrlichia spp., Bartonella coopersplainsensis, Bartonella tribocorum, Bartonella rattimassiliensis, Bartonella silvatica, two uncultured Bartonella spp., Babesia microti and diverse Hepatozoon were identified in six rodent species. More importantly, six species (including two Anaplasma, two Bartonella, "Ca. N. mikurensis" and Bab. microti) are zoonotic pathogens except Anaplasma bovis and Anaplasma ovis with zoonotic potential. Furthermore, dual infection was observed between different microorganisms, and the most common type of co-infection is between "Ca. N. mikurensis" and other microorganisms. Additionally, potential novel Bartonella species and Hepatozoon species demonstrated the presence of more diverse rodent-associated Bartonella and Hepatozoon. CONCLUSIONS: The results in this work indicated great genetic diversity of vector-borne infections in wild rodents, and highlighted the potential risk of human pathogens transmitted from rodents to humans through vectors.


Subject(s)
Genetic Variation , Rodentia , Animals , China/epidemiology , Rodentia/microbiology , Rodentia/parasitology , Phylogeny , Animals, Wild/parasitology , Animals, Wild/microbiology , Anaplasma/genetics , Anaplasma/isolation & purification , Anaplasma/classification , Vector Borne Diseases/transmission , Vector Borne Diseases/microbiology , Vector Borne Diseases/parasitology , Vector Borne Diseases/epidemiology , Bartonella/genetics , Bartonella/isolation & purification , Bartonella/classification , Bacteria/genetics , Bacteria/isolation & purification , Bacteria/classification , Rats
2.
Front Microbiol ; 15: 1359797, 2024.
Article in English | MEDLINE | ID: mdl-38605713

ABSTRACT

Rodents have been confirmed as hosts of various vector-borne zoonotic pathogens and are important for the maintenance of these microbes in nature. However, surveillance for zoonotic pathogens is limited for many wild rodent species in China, so our knowledge of pathogen ecology, genetic diversity, and the risk of cross-species transmission to humans is limited. In this study, 165 spleen samples of Daurian ground squirrels (Spermophilus dauricus) were collected from Weichang Manchu and the Mongolian Autonomous County of Hebei Province, China, and Rickettsia, Bartonella, and Anaplasma were identified by DNA detection using polymerase chain reaction (PCR). Sequence analysis identified eight bacterial pathogens: R. raoultii, R. sibirica, Candidatus R. longicornii, B. washoensis, B. grahamii, B. jaculi, A. capra, and Candidatus Anaplasma cinensis. Co-infection of B. grahamii and R. raoultii in one sample was observed. Our results demonstrated the genetic diversity of bacteria in Daurian ground squirrels and contributed to the distribution of these pathogens. Six species, A. capra, R. raoultii, R. sibirica, Candidatus R. longicornii, B. washoensis, and B. grahamii, are known to be pathogenic to humans, indicating a potential public health risk to the local human population, especially to herders who frequently have close contact with Daurian ground squirrels and are thus exposed to their ectoparasites.

3.
Front Vet Sci ; 11: 1373178, 2024.
Article in English | MEDLINE | ID: mdl-38550787

ABSTRACT

Background: Dogs and cats are the hosts of many vector-borne human pathogens that can be transmitted to humans. Given their direct and intimate contact with humans, companion dogs and cats are considered direct sentinels of vector-borne human pathogens. However, limited information is currently available regarding canine and feline zoonotic pathogens in China. This study detected canine and feline vector-borne human pathogens to better understand the potential risk to humans. Methods: Blood samples were collected from 275 domestic companion animals (117 dogs and 158 cats) living in Tianjin city, China, and the presence of DNA from Anaplasma, Babesia, Bartonella, and Rickettsia was detected by semi-nested polymerase chain reaction (PCR). The PCR products of the expected size were sequenced, and these newly generated sequences were subjected to BLASTN, nucleotide identity, and phylogenetic analyses. Results: A total of 24 blood samples tested positive for vector-borne pathogens in companion dogs and cats in Tianjin city, China, with a relatively low positive rate of 8.7%. Specifically, seven human pathogens, including Rickettsia raoultii, Candidatus Rickettsia jingxinensis, Rickettsia sibirica, Rickettsia felis, Babesia venatorum, Bartonella tribocorum, and Bartonella Henselae, were identified. In addition, Anaplasma ovis with zoonotic potential and Candidatus A. cinensis were detected. Conclusion: Our results indicate substantial genetic diversity in the vector-borne human pathogens circulating in companion dogs and cats. Interventions based on "One Health" should be taken to reduce the potential risks of contracting infection from companion dogs and cats in Tianjin, China.

4.
Front Immunol ; 14: 1187035, 2023.
Article in English | MEDLINE | ID: mdl-37207203

ABSTRACT

Host immune activation is critical for enterovirus 71 (EV71) clearance and immunopathogenesis. However, the mechanism of innate immune activation, especially of cell membrane-bound toll-like receptors (TLRs), against EV71 remains unknown. We previously demonstrated that TLR2 and its heterodimer inhibit EV71 replication. In this study, we systematically investigated the effects of TLR1/2/4/6 monomers and TLR2 heterodimer (TLR2/TLR1, TLR2/TLR6, and TLR2/TLR4) on EV71 replication and innate immune activation. We found that the overexpression of human- or mouse-derived TLR1/2/4/6 monomers and TLR2 heterodimer significantly inhibited EV71 replication and induced the production of interleukin (IL)-8 via activation of the phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) and mitogen-activated protein kinase (MAPK) pathways. Furthermore,human-mouse chimeric TLR2 heterodimer inhibited EV71 replication and activated innate immunity. Dominant-negative TIR-less (DN)-TLR1/2/4/6 did not exert any inhibitory effects, whereas DN-TLR2 heterodimer inhibited EV71 replication. Prokaryotic expression of purified recombinant EV71 capsid proteins (VP1, VP2, VP3, and VP4) or overexpression of EV71 capsid proteins induced the production of IL-6 and IL-8 via activation of the PI3K/AKT and MAPK pathways. Notably, two types of EV71 capsid proteins served as pathogen-associated molecular patterns for TLR monomers (TLR2 and TLR4) and TLR2 heterodimer (TLR2/TLR1, TLR2/TLR6, and TLR2/TLR4) and activated innate immunity. Collectively, our results revealed that membrane TLRs inhibited EV71 replication via activation of the antiviral innate response, providing insights into the EV71 innate immune activation mechanism.


Subject(s)
Enterovirus A, Human , Toll-Like Receptor 1 , Humans , Animals , Mice , Toll-Like Receptor 2/metabolism , Proto-Oncogene Proteins c-akt , Phosphatidylinositol 3-Kinases , Toll-Like Receptor 6/metabolism , Toll-Like Receptor 4 , Capsid Proteins , Toll-Like Receptors , Cell Membrane/metabolism , Antiviral Agents
5.
Front Microbiol ; 13: 1039665, 2022.
Article in English | MEDLINE | ID: mdl-36504836

ABSTRACT

Rodents are the primary natural reservoirs of Bartonella spp., and some of which are zoonotic causative agents. Hence, surveillance of Bartonella sp. infection in rodents is very important for the prevention of human bartonellosis caused by them. In this study, rodents were captured, and their spleen samples were collected for Bartonella sp. DNA detection and identification by amplifying the 16S rRNA, gltA, and ftsz genes using semi-nested polymerase chain reaction (PCR). The results indicated that Bartonella sp. DNA was detected in seven Rattus norvegicus individuals with a detection rate of 6.7% in Chengde City and bacterial DNA in 31 Apodemus agrarius individuals with a detection rate of 28.4% in Handan City. The DNA detection rate across the genders and ages of rodents was not found to be statistically significant. Furthermore, sequence analysis of the above-mentioned three genes demonstrated that at least eight Bartonella species were circulating in Hebei Province, of which three, including Bartonella rattimassiliensis, Bartonella grahamii, and Bartonella tribocorum, are human pathogens, thus suggesting the existence of a major public health risk. Overall, these results revealed the detection rate and genetic diversity of Bartonella species infection in rodents in Hebei Province, which could be potentially helpful for the prevention of bartonellosis caused by rodent-associated Bartonella species. This study highlights the urgent need for the surveillance of Bartonella infections in rodents and ectoparasites that affect both rodents and humans and can cause fever of unknown origin or endocarditis.

6.
J Microbiol Methods ; 202: 106601, 2022 11.
Article in English | MEDLINE | ID: mdl-36243230

ABSTRACT

"Candidatus Rickettsia tarasevichiae" (CRT) is increasingly being recognized as a disease causative agent in China and poses a great challenge to public health. Rapid and accurate detection is indispensable for laboratory diagnosis of infection caused by CRT and its surveillance in ticks. In the present study, a novel DNA-based loop-mediated isothermal amplification (LAMP) assay targeting the ompA gene was developed for the detection of CRT in tick samples. A set of universal primers specific to CRT were designed using PrimerExplorer V5 software. The analytical sensitivity, evaluated using recombinant plasmids containing the ompA gene, reached up to 1 copy per reaction, greater than that of the PCR assay targeting the same gene. This LAMP assay specifically detected CRT and showed no cross-reaction with other species common in China within the genus Rickettsia. In addition, this newly developed LAMP assay presented high diagnostic sensitivity of CRT detection validated by known positive DNA samples from ticks and simulated clinical samples. The applicability of the LAMP assay was evaluated by screening CRT from ticks, and the result showed that CRT circulation in Weichang County, China, was confirmed. Our findings indicate that this LAMP method is sensitive and specific for the detection of CRT and may have a potential application in the detection of CRT infection in patients and ticks.


Subject(s)
Rickettsia , Ticks , Animals , Humans , Rickettsia/genetics , Nucleic Acid Amplification Techniques/methods , Molecular Diagnostic Techniques/methods , Ticks/microbiology , Sensitivity and Specificity
7.
Medicine (Baltimore) ; 100(5): e23786, 2021 Feb 05.
Article in English | MEDLINE | ID: mdl-33592835

ABSTRACT

ABSTRACT: Community acquired-pneumonia (CAP) has varying causative pathogens and clinical characteristics. This study investigated the prevalence of Mycoplasma pneumoniae (M pneumoniae) and evaluated the clinical characteristics in infected hospitalized children by disease severity.From throat swabs of hospitalized children (5 months to 14 years) with CAP collected between November 2017 and May 2018, M pneumoniae and other CAP pathogens were identified using polymerase chain reaction (PCR). Differences in clinical and laboratory test data were compared between severe and mild case groups.Of 333 hospitalized children enrolled, 221/333 (66.4%) tested positive for M pneumoniae and 24/221 (10.9%) patients were (n = 9, aged <5 years vs n = 15, ≥5 years) single infection by PCR, however, only 170/333 (51.1%) patients were presented with M pneumoniae IgM-positive. M pneumoniae detection rate by PCR was higher than by immunoglobulin (IgM) serology. In 123/221 (55.7%) M pneumoniae infected patients, coinfection with bacterial pathogens (n = 61, <5 years vs n = 62, ≥5 years) occurred. Children (aged 3-8 years) had most M pneumoniae infection. Severe M pneumoniae pneumonia (MPP) in children occurred mostly in older age (7 [interquartile ranges {IQR}, 6-8] years; P < .0001), with longer cough days (14 [IQR, 10-19.5] days; P = .002) and hospitalization duration (9.5 [IQR, 7-12.3] days; P < .0001), lower lymphocyte ratio (24.1, [IQR, 20.0-31.1] %; P = .001), higher neutrophils ratio (66.0, [IQR, 60.2-70.3]%; P < .0001), and serum C-reactive protein (CRP) level (3.8, [IQR, 1.3-10.9] mg/L; P = .027).M pneumoniae is the most commonly detected pathogen in CAP. High coinfection prevalence increases diagnosis difficulty by clinically nonspecific characteristics. M pneumoniae detection by PCR with IgM may improve precise and reliable diagnosis of community-acquired MPP.


Subject(s)
Child, Hospitalized/statistics & numerical data , Community-Acquired Infections/epidemiology , Mycoplasma pneumoniae/isolation & purification , Pneumonia, Mycoplasma/epidemiology , Adolescent , Antibodies, Bacterial/blood , Antibodies, Bacterial/immunology , Child , Child, Preschool , China/epidemiology , Community-Acquired Infections/microbiology , Female , Humans , Immunoglobulin M/blood , Immunoglobulin M/immunology , Infant , Male , Mycoplasma pneumoniae/immunology , Pneumonia, Mycoplasma/microbiology , Polymerase Chain Reaction , Prevalence
8.
PLoS Negl Trop Dis ; 14(10): e0008778, 2020 10.
Article in English | MEDLINE | ID: mdl-33075097

ABSTRACT

In recent years, hemorrhagic fever with renal syndrome (HFRS) incidence has been becoming a severe public health problem again due to its significant increase in Shaanxi Province, China. Baoji, located in the Guanzhong Plain in the central part of Shaanxi Province, has been severely affected by HFRS since its first emergence in 1955. To better understand the epidemiology of orthohantaviruses infection in humans and the causative agents carried by the rodents, the long-term incidence patterns were analyzed and a molecular epidemiological investigation of orthohantaviruses infection in humans and rodents was performed. During 1984-2019, 13,042 HFRS cases were registered in Baoji, including 275 death cases. Except the first high prevalence of HFRS in 1988-1993, another two epidemic peaks were observed in 1998-2003 and 2012, respectively, although vaccination project was started since 1996. During the same period, HFRS cases in Baoji mainly were recorded in winter suggesting they may be caused by Hantaan orthohantavirus (HTNV), while a small peak of HFRS was also found in summer with unknown reason. Nucleotide identity and phylogenetic analyses demonstrated that a novel clade of HTNV sequences recovered from HFRS cases were closely related to those from rodents, including species close contact with humans, suggesting a direct viral transmission from rodents to humans and the important role in the HTNV transmission the nontraditional rodent hosts may play. Moreover, two distant related Dabieshan orthohantavirus (DBSV) lineages were also identified in Niviventer niviventer in this area demonstrating its considerable genetic diversity. Our data indicated that continual spillover of HTNV from rodents to humans, contributing to the high prevalence of HFRS in humans in Baoji.


Subject(s)
Hantaan virus/isolation & purification , Hemorrhagic Fever with Renal Syndrome/veterinary , Hemorrhagic Fever with Renal Syndrome/virology , Rodent Diseases/virology , Animals , China/epidemiology , Hantaan virus/classification , Hantaan virus/genetics , Hantaan virus/physiology , Hemorrhagic Fever with Renal Syndrome/epidemiology , Hemorrhagic Fever with Renal Syndrome/transmission , Humans , Incidence , Phylogeny , Rodent Diseases/epidemiology , Rodent Diseases/transmission , Rodentia/classification , Rodentia/virology , Seasons
10.
Virology ; 548: 213-225, 2020 09.
Article in English | MEDLINE | ID: mdl-32763492

ABSTRACT

The alteration of host cell splicing is a major strategy favouring viral replication; however, the interaction between human tonsillar epithelial cells (HTECs) and enterovirus 71 (EV71) has not been fully elucidated. Here, a total of 201 differentially expressed genes (DEGs) and 3266 novel genes with coding potential were identified. A total of 3479 skipped exons (SEs), 515 alternative 3' splice sites (A3SSs), 391 alternative 5' splice sites (A5SSs), 531 mutually exclusive exons (MXEs) and 825 retained introns (RIs) were identified as significantly altered alternative splicing (AS) events. The enriched DEGs were mainly related to the cell cycle, spliceosome, and Toll-like receptor (TLR) signalling pathways. Finally, the replication of EV71 was significantly inhibited by TLR2 heterodimers. Our findings suggest that AS events induced by EV71 increase the transcriptomic diversity of HTECs in response to EV71 infection. Additionally, TLR2 heterodimers have the potential to protect HTECs against EV71.


Subject(s)
Enterovirus A, Human/physiology , Enterovirus Infections/genetics , Alternative Splicing , Enterovirus A, Human/genetics , Enterovirus Infections/metabolism , Enterovirus Infections/virology , Epithelial Cells/metabolism , Epithelial Cells/virology , Host-Pathogen Interactions , Humans , RNA Splice Sites , Transcriptome
11.
Parasit Vectors ; 13(1): 366, 2020 Jul 22.
Article in English | MEDLINE | ID: mdl-32698848

ABSTRACT

BACKGROUND: Several members of genus Babesia are important pathogens causing babesiosis in dogs. In China, at least five Babesia species have been described in dogs or ticks. This study sought to determine the prevalence and molecular characteristics of various Babesia spp. in dogs in cities in Shaanxi Province in China, including Xi'an and Hanzhong. METHODS: A total of 371 blood samples were collected from pet dogs presenting to veterinary clinics in the cities of Xi'an and Hanzhong in Shaanxi, China. Babesia spp. DNA was detected via amplification of partial 18S rRNA genes by semi-nested PCR. Almost full-length 18S rRNA, ITS, partial TRAP and complete cytb genes were recovered for analysis of the genetic characteristics and relationships with known isolates. RESULTS: A single species, Babesia gibsoni, was identified in dogs in Xi'an and Hanzhong. Consistently, B. gibsoni was also detected in 14 ticks collected from positive dogs. Sequence similarities and phylogenetic analysis suggested that the isolates identified herein showed a closer genetic relationship with isolates from East Asian countries rather than India, Bangladesh, or the USA. Sequence analysis based on tandem repeat analysis of the TRAP gene further revealed that specific haplotypes were circulating in both Xi'an and Hanzhong, with no specific regionality. In addition, 10.9% of all isolates with atovaquone (ATV)-resistance were identified because of M121I mutation in the deduced cytb protein. CONCLUSIONS: This study revealed a high prevalence rate of Babesia infection. Babesia gibsoni was the only Babesia species identified in cases of canine babesiosis in the cities of Xi'an and Hanzhong cities in Shaanxi, China. In addition, the TRAP gene presented high genetic diversity across isolates. Such information is useful for elucidating the epidemiological characteristics of canine babesiosis, as well as the overall genetic diversity of Babesia spp. circulating in dog populations in Shaanxi Province.


Subject(s)
Babesia , Babesiosis , Animals , Arachnid Vectors/parasitology , Atovaquone/pharmacology , Babesia/genetics , Babesia/isolation & purification , Babesiosis/epidemiology , Babesiosis/transmission , Blood/parasitology , China/epidemiology , Dog Diseases/epidemiology , Dog Diseases/transmission , Dogs , Drug Resistance/genetics , Genes, Protozoan , Genetic Variation , Phylogeny , Polymerase Chain Reaction/veterinary , RNA, Ribosomal, 18S/genetics , Ticks/parasitology
12.
Ticks Tick Borne Dis ; 11(5): 101477, 2020 09.
Article in English | MEDLINE | ID: mdl-32723632

ABSTRACT

Anaplasma bovis is an organism significant to cattle and buffalo since it is one of the causative agents of bovine anaplasmosis. Previous studies have shown the worldwide distribution of A. bovis. However, most of these studies about its genetic diversity only focused on the rrs gene. In this study, DNA of A. bovis was detected in blood samples of cattle and goats in Xi'an city, China by nested-PCR. Near full-length rrs, groEL, and gltA genes were amplified successfully from the positive samples. Genetic analysis showed that specific genetic marker (an insertion and a deletion) was found in the rrs sequences in some strains, as well as clone 88 from monkeys in previous study. Phylogenetic analysis based on the rrs, groEL, and gltA genes revealed that A. bovis circulating in Xi'an exhibited great genetic diversity. Our results also indicated that variants outside China presented geographic clustering, and all A. bovis isolates based on the groEL or gltA gene also showed a host origin clustering. Also of note was that the phylogenetic analyses of the groEL and gltA genes suggested that both frequent dispersals over long distances in recent years and local adaptation over long evolutionary timescales played important roles in the distribution and evolution of A. bovis in China. Finally, a potential recombination event in the genome of Zhouzhi-cattle-10 based on inconsistent positions in the groEL and gltA trees was also observed. These results also reinforce the need for assessing the pathogenicity to humans of A. bovis variants with specific marker in the rrs gene.


Subject(s)
Anaplasma/genetics , Anaplasmosis/epidemiology , Cattle Diseases/epidemiology , Genetic Variation , Goat Diseases/epidemiology , Sheep Diseases/epidemiology , Anaplasma/isolation & purification , Anaplasmosis/microbiology , Animals , Base Sequence , Cattle , Cattle Diseases/microbiology , China/epidemiology , Genes, Bacterial , Goat Diseases/microbiology , Goats , Phylogeny , Sequence Alignment/veterinary , Sheep , Sheep Diseases/microbiology , Sheep, Domestic
13.
Comp Immunol Microbiol Infect Dis ; 72: 101514, 2020 Oct.
Article in English | MEDLINE | ID: mdl-32634650

ABSTRACT

Hepatozoon canis, transmitted by Rhipicephalus sanguineus, is a tick-borne pathogen and causes canine hepatozoonosis. Until now, only limited previous studies were conducted on the molecular detection and characterization of Hepatozoon sp. in dogs in China. Blood samples were collected from 93 sick dogs that were clinically diagnosed as babesiosis but tested negative for Babesia, and 103 apparently healthy dogs, as well as their infesting ticks in Xi'an and Hanzhong cities, Shaanxi province of China. PCR amplifying partial 18S rRNA gene was used to detect the DNA of Hepatozoon sp. Genetic and phylogenetic analysis were performed to determine the Hepatozoon species. Our results demonstrated that H. canis was identified from the sick dogs and the infested ticks in Hanzhong, with no significant differences of prevalence between both genders and ages. No positive blood or tick samples were found in Xi'an. Moreover, all the 18S rRNA gene sequences recovered from both dogs and the infested ticks showed a high genetic similarity with each other, and also presented a close relationship with other known sequences in and outside China. In conclusion, H. canis was identified in babesiosis-suspected dogs and ticks infesting them in Shaanxi, China, although the association between clinical signs and H. canis need further study.


Subject(s)
Coccidiosis/veterinary , Dog Diseases , Eucoccidiida , Animals , China/epidemiology , Coccidiosis/epidemiology , Dog Diseases/diagnosis , Dog Diseases/epidemiology , Dog Diseases/parasitology , Dogs , Eucoccidiida/genetics , Eucoccidiida/isolation & purification , Female , Male , Phylogeny
14.
Int J Infect Dis ; 96: 538-540, 2020 Jul.
Article in English | MEDLINE | ID: mdl-32439540

ABSTRACT

OBJECTIVES: To better understand the spectrums of pathogens causing herpangina and circulation of Coxsackievirus A4 in Yancheng, China. METHODS: Stool samples from herpangina and HFMD cases were collected. Real Time PCR Kits was used to identify Enterovirus 71, CV-A16 and CV-A6, and nested reverse transcription PCR (nRT-PCR) to detect the other enterovirus types. Complete VP1 and genome sequence of CV-A4 were amplified by using nRT-PCR. Genetic, phylogenetic and recombination analysis were performed. RESULTS: Co-circulation of three recombinant CV-A4 groups, including one novel (C2 lineage), was identified in Yancheng, China, 2016 and 2018. One was the major causative agent of herpangina, and another two were responsible for HFMD. Phylogenetic and recombination analysis indicated that the non-structural region of their genome originated from the same ancestry and subsequently adaptation. C2 lineage of CV-A4 group may be introduced from countries outside China and its genome occurred recombination in China. CONCLUSION: Novel recombinant CV-A4 was mainly associated with herpanginain in Yancheng, 2018, China. C2 lineage of CV-A4 group with recombinant non-structural region was also identified in HFMD patients.


Subject(s)
Enterovirus A, Human/genetics , Enterovirus A, Human/isolation & purification , Enterovirus/isolation & purification , Genome, Viral , Hand, Foot and Mouth Disease/virology , Herpangina/virology , China/epidemiology , Enterovirus/classification , Enterovirus/genetics , Enterovirus A, Human/classification , Feces/virology , Genotype , Hand, Foot and Mouth Disease/epidemiology , Herpangina/epidemiology , Humans , Phylogeny , Real-Time Polymerase Chain Reaction , Recombination, Genetic
15.
Biomed Environ Sci ; 31(2): 136-145, 2018 02.
Article in English | MEDLINE | ID: mdl-29606192

ABSTRACT

OBJECTIVE: Newly identified human rhinovirus C (HRV-C) and human bocavirus (HBoV) cannot propagate in vitro in traditional cell culture models; thus obtaining knowledge about these viruses and developing related vaccines are difficult. Therefore, it is necessary to develop a novel platform for the propagation of these types of viruses. METHODS: A platform for culturing human airway epithelia in a three-dimensional (3D) pattern using Matrigel as scaffold was developed. The features of 3D culture were identified by immunochemical staining and transmission electron microscopy. Nucleic acid levels of HRV-C and HBoV in 3D cells at designated time points were quantitated by real-time polymerase chain reaction (PCR). Levels of cytokines, whose secretion was induced by the viruses, were measured by ELISA. RESULTS: Properties of bronchial-like tissues, such as the expression of biomarkers CK5, ZO-1, and PCK, and the development of cilium-like protuberances indicative of the human respiration tract, were observed in 3D-cultured human airway epithelial (HAE) cultures, but not in monolayer-cultured cells. Nucleic acid levels of HRV-C and HBoV and levels of virus-induced cytokines were also measured using the 3D culture system. CONCLUSION: Our data provide a preliminary indication that the 3D culture model of primary epithelia using a Matrigel scaffold in vitro can be used to propagate HRV-C and HBoV.


Subject(s)
Enterovirus Infections/virology , Enterovirus/growth & development , Human bocavirus/growth & development , Parvoviridae Infections/virology , Primary Cell Culture/methods , Respiratory Mucosa/virology , Collagen , Drug Combinations , Enterovirus/isolation & purification , Enzyme-Linked Immunosorbent Assay , Epithelial Cells/virology , Human bocavirus/isolation & purification , Humans , Laminin , Proteoglycans , Real-Time Polymerase Chain Reaction , Virus Cultivation
16.
Arch Virol ; 163(5): 1187-1193, 2018 May.
Article in English | MEDLINE | ID: mdl-29387970

ABSTRACT

To establish an animal model for the newly identified Marmota Himalayana hepatovirus, MHHAV, so as to develop a better understanding of the infection of hepatitis A viruses. Five experimental woodchucks (Marmota monax) were inoculated intravenously with the purified MHHAV from wild woodchuck feces. One animal injected with PBS was defined as a control. Feces and blood were routinely collected. After the animals were subjected to necropsy, different tissues were collected. The presence of viral RNA and negative sense viral RNA was analyzed in all the samples and histopathological and in situ hybridization analysis was performed for the tissues. MHHAV infection caused fever but no severe symptoms or death. Virus was shed in feces beginning at 2 dpi, and MHHAV RNA persisted in feces for ~2 months, with a biphasic increase, and in blood for ~30 days. Viral RNA was detected in all the tissues, with high levels in the liver and spleen. Negative-strand viral RNA was detected only in the liver. Furthermore, the animals showed histological signs of hepatitis at 45 dpi. MHHAV can infect M. monax and is associated with hepatic disease. Therefore, this animal can be used as a model of HAV pathogenesis and to evaluate antiviral and anticancer therapeutics.


Subject(s)
Disease Models, Animal , Hepatitis A virus/pathogenicity , Hepatitis A , Hepatitis, Viral, Animal , Marmota , Animals , Feces/virology , Hepatitis A/pathology , Hepatitis A/physiopathology , Hepatitis A/virology , Hepatitis A virus/genetics , Hepatitis A virus/isolation & purification , Hepatitis A virus/physiology , Hepatitis, Viral, Animal/pathology , Hepatitis, Viral, Animal/physiopathology , Hepatitis, Viral, Animal/virology , Liver/pathology , Liver/virology , RNA, Viral/isolation & purification , Spleen/pathology , Spleen/virology
17.
Biochem Biophys Res Commun ; 490(1): 22-28, 2017 08 12.
Article in English | MEDLINE | ID: mdl-28576493

ABSTRACT

Rhinovirus C (RV-C), a newly identified group of human rhinoviruses (RVs), is associated with exacerbation of severe asthma. The type I interferon (IFN) response induced by this virus and the mechanisms of evasion of IFN-mediated innate immunity for RV-C remain unclear. In this study, we constructed a full-length cDNA clone of RV-C (LZ651) from a clinical sample. IFN-ß mRNA and protein levels were not elevated in differentiated Human bronchial epithelial (HBE) cells at the air-liquid interface infected with RV-C, except in the early stage of infection. The ability to attenuate IFN-ß activation was ascribed to 3Cpro of RV-C, and the 40-His site of 3Cpro played an important role. Furthermore, RIG-I was degraded by 3Cpro in a caspase-dependent manner and 3Cpro cleaved MAVS at 148 Q/A, which inhibited IFN signaling. Taken together, our results demonstrate the mechanism by which RV-C circumvents the production of type I IFN in infected cells.


Subject(s)
Immune Tolerance , Immunity, Innate/immunology , Rhinovirus/immunology , HEK293 Cells , HeLa Cells , Humans , Interferon Type I/immunology
18.
Biomed Environ Sci ; 29(4): 286-9, 2016 04.
Article in English | MEDLINE | ID: mdl-27241739

ABSTRACT

Norovirus (NoV) is a pathogen that commonly causes viral diarrhea in children. Studies indicate that NoV recognizes human histo-blood group antigens (HBGAs) as cell attachment factors. In order to explore the correlation between of NoV infection and HBGAs, a cross-sectional study was conducted in children less than five years old who were hospitalized with diarrhea in two areas of China between November 2014 and February 2015. Of the paired stool and saliva samples taken from 424 children, NoV was detected in 24 (6%) children, with viral genotypes GII.3 (n=5), GII.4 (n=14), GII.12 (n=1), and GII.17 (n=4). All of the individuals having NoV infection were either secretors (Lea-b+/Lex-y+) or partial secretors (Lea+b+/Lex+y+) except one GII.3 infection of a non-secretor (Lea+b-/Lex+y-). These results suggest that secretor positive is associated with NoV infection, although non-secretors are not absolutely protected from NoV infection.


Subject(s)
Blood Group Antigens/genetics , Caliciviridae Infections/blood , Caliciviridae Infections/complications , Diarrhea/blood , Diarrhea/etiology , Gastroenteritis/blood , Norovirus/physiology , Caliciviridae Infections/virology , Child, Preschool , China , Cross-Sectional Studies , Diarrhea/virology , Feces/virology , Gastroenteritis/virology , Genotype , Humans , Infant
19.
Virology ; 494: 108-18, 2016 07.
Article in English | MEDLINE | ID: mdl-27107253

ABSTRACT

A recent histopathologic study implicated human tonsillar crypt epithelium as an important site for EV71 replication in EV71-caused fatal cases. This study aimed to confirm the susceptibility of human tonsillar epithelium to EV71. Two human tonsillar epithelial cell lines (UT-SCC-60A and UT-SCC-60B) were susceptive to EV71, and PI3K/AKT, p38, ERK1/2, and JNK1/2 signal pathways were activated. Interferon-α, IL-8, IL-1ß, IL-6 and IL-12p40 were induced and regulated by PI3K/AKT, p38, ERK1/2, and JNK1/2 signal pathways. PI3K/AKT pathway activation appeared to suppress the induction of TNF-α, which induced cell survival by inhibiting GSK-3ß. The activation of NF-κB was observed but inhibited by these pathways in EV71 infection. Furthermore, ERK1/2 and JNK1/2 were essential for efficient EV71 replication. Human tonsillar epithelial cells support EV71 replication and display innate antiviral immunity in vitro, indicating that human tonsillar epithelial cells may be novel targets for EV71 infection and replication in vivo.


Subject(s)
Cytokines/biosynthesis , Enterovirus A, Human/physiology , Enterovirus Infections/metabolism , Enterovirus Infections/virology , Epithelial Cells/metabolism , Epithelial Cells/virology , Palatine Tonsil/metabolism , Palatine Tonsil/virology , Animals , Biomarkers , Cell Line , Cytokines/genetics , Cytopathogenic Effect, Viral , Disease Susceptibility , Epithelial Cells/pathology , Host-Pathogen Interactions , Humans , Inflammation Mediators/metabolism , Keratins/metabolism , Lysosomal Membrane Proteins/metabolism , Membrane Glycoproteins/metabolism , Mice , Mitogen-Activated Protein Kinases/metabolism , Mutation , NF-kappa B/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , RNA, Viral , Receptors, Scavenger/metabolism , Signal Transduction , Viral Proteins/genetics , Viral Proteins/metabolism , Virus Replication
20.
Biomed Environ Sci ; 29(2): 137-42, 2016 Feb.
Article in English | MEDLINE | ID: mdl-27003171

ABSTRACT

The aim of this study was to investigate the knockdown efficiency of 2'-O-methylated (2'-OMe)-modified small interfering RNAs (siRNAs) on human rhinovirus 1B (HRV1B) replication and the interferon response. Thus, 24 2'-OMe-modified siRNAs were designed to target HRV1B. The RNA levels of HRV1B, Toll-like receptor 3, melanoma differentiation-associated gene 5, retinoic acid inducible gene-I, and interferons were determined in HRV1B-infected HeLa and BEAS-2B epithelial cells transfected with 2'-OMe-modified siRNAs. The results revealed that all 2'-OMe-modified siRNAs interfered with the replication of HRV1B in a cell-specific and transfection efficiency-dependent manner. Viral activation of Toll-like receptor 3, melanoma differentiation-associated gene 5, retinoic acid inducible gene-I, and the interferon response was detected. In conclusion, the 2'-OMe-modified siRNAs used in this study could interfere with HRV1B replication, possibly leading to the reactivation of the interferon response.


Subject(s)
Gene Knockdown Techniques , Rhinovirus , HeLa Cells , Humans , Interferons/physiology , RNA, Small Interfering , Virus Replication
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