24 results on '"Miaoge Xue"'
Search Results
2. Viral RNA N6-methyladenosine modification modulates both innate and adaptive immune responses of human respiratory syncytial virus.
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Miaoge Xue, Yuexiu Zhang, Haitao Wang, Elizabeth L Kairis, Mijia Lu, Sadeem Ahmad, Zayed Attia, Olivia Harder, Zijie Zhang, Jiangbo Wei, Phylip Chen, Youling Gao, Mark E Peeples, Amit Sharma, Prosper Boyaka, Chuan He, Sun Hur, Stefan Niewiesk, and Jianrong Li
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
Human respiratory syncytial virus (RSV) is the leading cause of respiratory tract infections in humans. A well-known challenge in the development of a live attenuated RSV vaccine is that interferon (IFN)-mediated antiviral responses are strongly suppressed by RSV nonstructural proteins which, in turn, dampens the subsequent adaptive immune responses. Here, we discovered a novel strategy to enhance innate and adaptive immunity to RSV infection. Specifically, we found that recombinant RSVs deficient in viral RNA N6-methyladenosine (m6A) and RSV grown in m6A methyltransferase (METTL3)-knockdown cells induce higher expression of RIG-I, bind more efficiently to RIG-I, and enhance RIG-I ubiquitination and IRF3 phosphorylation compared to wild-type virion RNA, leading to enhanced type I IFN production. Importantly, these m6A-deficient RSV mutants also induce a stronger IFN response in vivo, are significantly attenuated, induce higher neutralizing antibody and T cell immune responses in mice and provide complete protection against RSV challenge in cotton rats. Collectively, our results demonstrate that inhibition of RSV RNA m6A methylation enhances innate immune responses which in turn promote adaptive immunity.
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- 2021
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3. A Zika virus vaccine expressing premembrane-envelope-NS1 polyprotein
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Anzhong Li, Jingyou Yu, Mijia Lu, Yuanmei Ma, Zayed Attia, Chao Shan, Miaoge Xue, Xueya Liang, Kelsey Craig, Nirajkumar Makadiya, Jennifer J. He, Ryan Jennings, Pei-Yong Shi, Mark E. Peeples, Shan-Lu Liu, Prosper N. Boyaka, and Jianrong Li
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Science - Abstract
Current Zika virus (ZIKV) subunit vaccine development largely focuses on prM and E proteins, and the role of NS1 for immune response and protection is unclear. Here, Li et al. develop an attenuated VSV-based vaccine expressing a ZIKV prM-E-NS1 polyprotein and characterize immune response and protection in mice.
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- 2018
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4. A Lactic Acid Bacteria (LAB)-Based Vaccine Candidate for Human Norovirus
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Kelsey Craig, Xianjun Dai, Anzhong Li, Mijia Lu, Miaoge Xue, Lucia Rosas, Thomas Z. Gao, Andrew Niehaus, Ryan Jennings, and Jianrong Li
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human norovirus ,lactic acid bacteria ,gnotobiotic piglet ,vaccine ,VP1 ,Microbiology ,QR1-502 - Abstract
Human noroviruses (HuNoVs) are responsible for more than 95% of the non-bacterial acute gastroenteritis epidemics in the world. The CDC estimates that every year 21 million individuals suffer from HuNoV-induced gastroenteritis in the United States. Currently, there is no FDA-approved vaccine for HuNoVs. Development of an effective vaccine has been hampered by the lack of an efficient cell culture system for HuNoVs and a suitable small animal model for pathogenesis study. In this study, we developed lactic acid bacteria (LAB) as a vector to deliver HuNoV antigen. A LAB strain (Lactococcus lactis) carrying VP1 gene of a HuNoV GII.4 virus (LAB-VP1) was constructed. It was found that HuNoV VP1 protein was highly expressed by LAB vector and was secreted into media supernatants. To test whether LAB-based HuNoV vaccine candidate is immunogenic, 4-day-old gnotobiotic piglets were orally inoculated with various doses of LAB-VP1. It was found that LABs were persistent in the small intestine of piglets and shed in pig feces for at least 25 days post inoculation. LAB DNA and VP1 were detected in mesenteric lymph nodes and spleen tissue in LAB-VP1 inoculated groups. HuNoV-specific IgG and IgA were detectable in serum and feces respectively at day 13 post-inoculation, and further increased at later time points. After being challenged with HuNoV GII.4 strain, a large amount of HuNoV antigens were observed in the duodenum, jejunum, and ileum sections of the intestine in the LAB control group. In contrast, significantly less, or no, HuNoV antigens were detected in the LAB-VP1 immunized groups. Collectively, these results demonstrate that a LAB-based HuNoV vaccine induces protective immunity in gnotobiotic piglets.
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- 2019
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- View/download PDF
5. N6-methyladenosine modification enables viral RNA to escape recognition by RNA sensor RIG-I
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Yunsheng Xu, Chuan He, Mark E. Peeples, Olivia Harder, Xueya Liang, Boxuan Simen Zhao, Jianrong Li, Zongdi Feng, Miaoge Xue, Jiyong Zhou, Mijia Lu, Thomas Z. Gao, Zijie Zhang, Stefan Niewiesk, and Anzhong Li
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Microbiology (medical) ,viruses ,Immunology ,Applied Microbiology and Biotechnology ,Microbiology ,03 medical and health sciences ,chemistry.chemical_compound ,Human metapneumovirus ,Interferon ,Gene expression ,Genetics ,medicine ,030304 developmental biology ,Regulation of gene expression ,0303 health sciences ,biology ,030306 microbiology ,Chemistry ,RIG-I ,virus diseases ,RNA ,MDA5 ,Cell Biology ,biology.organism_classification ,Cell biology ,N6-Methyladenosine ,medicine.drug - Abstract
Internal N6-methyladenosine (m6A) modification is one of the most common and abundant modifications of RNA. However, the biological roles of viral RNA m6A remain elusive. Here, using human metapneumovirus (HMPV) as a model, we demonstrate that m6A serves as a molecular marker for innate immune discrimination of self from non-self RNAs. We show that HMPV RNAs are m6A methylated and that viral m6A methylation promotes HMPV replication and gene expression. Inactivating m6A addition sites with synonymous mutations or demethylase resulted in m6A-deficient recombinant HMPVs and virion RNAs that induced increased expression of type I interferon, which was dependent on the cytoplasmic RNA sensor RIG-I, and not on melanoma differentiation-associated protein 5 (MDA5). Mechanistically, m6A-deficient virion RNA induces higher expression of RIG-I, binds more efficiently to RIG-I and facilitates the conformational change of RIG-I, leading to enhanced interferon expression. Furthermore, m6A-deficient recombinant HMPVs triggered increased interferon in vivo and were attenuated in cotton rats but retained high immunogenicity. Collectively, our results highlight that (1) viruses acquire m6A in their RNA as a means of mimicking cellular RNA to avoid detection by innate immunity and (2) viral RNA m6A can serve as a target to attenuate HMPV for vaccine purposes. This study reports a novel function for the N6-methyladenosine RNA modification in allowing RIG-I to discriminate self from non-self RNA and shows that human metapneumovirus induces this modification of its RNA to evade recognition in vivo.
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- 2020
6. Nonsegmented Negative-Sense RNA Viruses Utilize N(6)-Methyladenosine (m(6)A) as a Common Strategy To Evade Host Innate Immunity
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Qinzhe Liu, Jiangbo Wei, Sadeem Ahmad, Mark E. Peeples, Amit Sharma, Jianrong Li, Elizabeth L. Kairis, Chuan He, Youling Gao, Dominique Garcin, Sun Hur, Mijia Lu, Miaoge Xue, Yuexiu Zhang, Hai-Tao Wang, and Zijie Zhang
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Gene Expression Regulation, Viral ,Adenosine ,viruses ,Immunology ,Microbiology ,03 medical and health sciences ,chemistry.chemical_compound ,Gene Knockout Techniques ,0302 clinical medicine ,RNA Virus Infections ,Interferon ,Virology ,Gene expression ,medicine ,Humans ,RNA Processing, Post-Transcriptional ,030304 developmental biology ,Genetics ,0303 health sciences ,Innate immune system ,biology ,Host Microbial Interactions ,RNA ,Methyltransferases ,Rhabdoviridae ,biology.organism_classification ,Immunity, Innate ,Pneumoviridae ,chemistry ,Type I interferon signaling pathway ,A549 Cells ,Insect Science ,Interferon Type I ,Negative-Sense RNA Viruses ,Pathogenesis and Immunity ,RNA, Viral ,N6-Methyladenosine ,030217 neurology & neurosurgery ,medicine.drug - Abstract
N(6)-Methyladenosine (m(6)A) is the most abundant internal RNA modification catalyzed by host RNA methyltransferases. As obligate intracellular parasites, many viruses acquire m(6)A methylation in their RNAs. However, the biological functions of viral m(6)A methylation are poorly understood. Here, we found that viral m(6)A methylation serves as a molecular marker for host innate immunity to discriminate self from nonself RNA and that this novel biological function of viral m(6)A methylation is universally conserved in several families in nonsegmented negative-sense (NNS) RNA viruses. Using m(6)A methyltransferase (METTL3) knockout cells, we produced m(6)A-deficient virion RNAs from the representative members of the families Pneumoviridae, Paramyxoviridae, and Rhabdoviridae and found that these m(6)A-deficient viral RNAs triggered significantly higher levels of type I interferon compared to the m(6)A-sufficient viral RNAs, in a RIG-I-dependent manner. Reconstitution of the RIG-I pathway revealed that m(6)A-deficient virion RNA induced higher expression of RIG-I, bound to RIG-I more efficiently, enhanced RIG-I ubiquitination, and facilitated RIG-I conformational rearrangement and oligomerization. Furthermore, the m(6)A binding protein YTHDF2 is essential for suppression of the type I interferon signaling pathway, including by virion RNA. Collectively, our results suggest that several families in NNS RNA viruses acquire m(6)A in viral RNA as a common strategy to evade host innate immunity. IMPORTANCE The nonsegmented negative-sense (NNS) RNA viruses share many common replication and gene expression strategies. There are no vaccines or antiviral drugs for many of these viruses. We found that representative members of the families Pneumoviridae, Paramyxoviridae, and Rhabdoviridae among the NNS RNA viruses acquire m(6)A methylation in their genome and antigenome as a means to escape recognition by host innate immunity via a RIG-I-dependent signaling pathway. Viral RNA lacking m(6)A methylation induces a significantly higher type I interferon response than m(6)A-sufficient viral RNA. In addition to uncovering m(6)A methylation as a common mechanism for many NNS RNA viruses to evade host innate immunity, this study discovered a novel strategy to enhance type I interferon responses, which may have important applications in vaccine development, as robust innate immunity will likely promote the subsequent adaptive immunity.
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- 2021
7. Stable Attenuation of Human Respiratory Syncytial Virus for Live Vaccines by Deletion and Insertion of Amino Acids in the Hinge Region between the mRNA Capping and Methyltransferase Domains of the Large Polymerase Protein
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Krista M. D. La Perle, Ryan N. Jennings, Stefan Niewiesk, Jianrong Li, Anzhong Li, Miaoge Xue, Rongzhang Wang, Phylip Chen, Xueya Liang, Mark E. Peeples, Hui Cai, Mijia Lu, and Olivia Harder
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Models, Molecular ,viruses ,Immunology ,Respiratory Syncytial Virus Infections ,Antibodies, Viral ,Vaccines, Attenuated ,Virus Replication ,Microbiology ,Virus ,Cell Line ,law.invention ,Viral Proteins ,03 medical and health sciences ,chemistry.chemical_compound ,law ,Virology ,RNA polymerase ,Vaccines and Antiviral Agents ,Chlorocebus aethiops ,Respiratory Syncytial Virus Vaccines ,Animals ,Humans ,RNA, Messenger ,Sigmodontinae ,Amino Acids ,Neutralizing antibody ,Lung ,Vero Cells ,Polymerase ,030304 developmental biology ,0303 health sciences ,Messenger RNA ,Innate immune system ,biology ,Immunogenicity ,030302 biochemistry & molecular biology ,Methyltransferases ,RNA-Dependent RNA Polymerase ,chemistry ,A549 Cells ,Respiratory Syncytial Virus, Human ,Insect Science ,biology.protein ,Recombinant DNA ,Cytokines - Abstract
Human respiratory syncytial virus (RSV) is the leading viral cause of lower respiratory tract disease in infants and children worldwide. Currently, there are no FDA-approved vaccines to combat this virus. The large (L) polymerase protein of RSV replicates the viral genome and transcribes viral mRNAs. The L protein is organized as a core ring-like domain containing the RNA-dependent RNA polymerase and an appendage of globular domains containing an mRNA capping region and a cap methyltransferase region, which are linked by a flexible hinge region. Here, we found that the flexible hinge region of RSV L protein is tolerant to amino acid deletion or insertion. Recombinant RSVs carrying a single or double deletion or a single alanine insertion were genetically stable, highly attenuated in immortalized cells, had defects in replication and spread, and had a delay in innate immune cytokine responses in primary, well-differentiated, human bronchial epithelial (HBE) cultures. The replication of these recombinant viruses was highly attenuated in the upper and lower respiratory tracts of cotton rats. Importantly, these recombinant viruses elicited high levels of neutralizing antibody and provided complete protection against RSV replication. Taken together, amino acid deletions or insertions in the hinge region of the L protein can serve as a novel approach to rationally design genetically stable, highly attenuated, and immunogenic live virus vaccine candidates for RSV. IMPORTANCE Despite tremendous efforts, there are no FDA-approved vaccines for human respiratory syncytial virus (RSV). A live attenuated RSV vaccine is one of the most promising vaccine strategies for RSV. However, it has been a challenge to identify an RSV vaccine strain that has an optimal balance between attenuation and immunogenicity. In this study, we generated a panel of recombinant RSVs carrying a single and double deletion or a single alanine insertion in the large (L) polymerase protein that are genetically stable, sufficiently attenuated, and grow to high titer in cultured cells, while retaining high immunogenicity. Thus, these recombinant viruses may be promising vaccine candidates for RSV.
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- 2020
8. Vesicular Stomatitis Virus and DNA Vaccines Expressing Zika Virus Nonstructural Protein 1 Induce Substantial but Not Sterilizing Protection against Zika Virus Infection
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Zayed Attia, Mark E. Peeples, Anzhong Li, Pei Yong Shi, Xueya Liang, Jingyou Yu, Mijia Lu, Shan-Lu Liu, Thomas Z. Gao, Miaoge Xue, Chao Shan, Jianrong Li, and Prosper N. Boyaka
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viruses ,Immunology ,Viremia ,Receptor, Interferon alpha-beta ,Cross Reactions ,Viral Nonstructural Proteins ,Antibodies, Viral ,Microbiology ,Vesicular stomatitis Indiana virus ,Zika virus ,Dengue fever ,DNA vaccination ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Virology ,Vaccines and Antiviral Agents ,Vaccines, DNA ,medicine ,Animals ,Antibody-dependent enhancement ,030212 general & internal medicine ,030304 developmental biology ,Mice, Knockout ,Mice, Inbred BALB C ,0303 health sciences ,biology ,Zika Virus Infection ,Immunogenicity ,virus diseases ,Viral Vaccines ,Zika Virus ,biochemical phenomena, metabolism, and nutrition ,biology.organism_classification ,medicine.disease ,Antibodies, Neutralizing ,Disease Models, Animal ,Flavivirus ,Vesicular stomatitis virus ,Insect Science ,Female ,Vesicular Stomatitis - Abstract
The nonstructural protein 1 (NS1) of several flaviviruses, including West Nile, dengue, and yellow fever viruses, is capable of inducing variable degrees of protection against flavivirus infection in animal models. However, the immunogenicity of NS1 protein of Zika virus (ZIKV) is less understood. Here, we determined the efficacy of ZIKV NS1-based vaccine candidates using two delivery platforms, methyltransferase-defective recombinant vesicular stomatitis virus (mtdVSV) and a DNA vaccine. We first show that expression of ZIKV NS1 could be significantly enhanced by optimizing the signal peptide. A single dose of mtdVSV-NS1-based vaccine or two doses of DNA vaccine induced high levels of NS1-specfic antibody and T cell immune responses but provided only partial protection against ZIKV viremia in BALB/c mice. In Ifnar1(−/−) mice, neither NS1-based vaccine provided protection against a lethal high dose (10(5) PFU) ZIKV challenge, but mtdVSV-NS1-based vaccine prevented deaths from a low dose (10(3) PFU) challenge, though they experienced viremia and body weight loss. We conclude that ZIKV NS1 alone conferred substantial, but not complete, protection against ZIKV infection. Nevertheless, these results highlight the value of ZIKV NS1 for vaccine development. IMPORTANCE Most Zika virus (ZIKV) vaccine research has focused on the E or prM-E proteins and the induction of high levels of neutralizing antibodies. However, these ZIKV neutralizing antibodies cross-react with other flaviviruses, which may aggravate the disease via an antibody-dependent enhancement (ADE) mechanism. ZIKV NS1 protein may be an alternative antigen for vaccine development, since antibodies to NS1 do not bind to the virion, thereby eliminating the risk of ADE. Here, we show that recombinant VSV and DNA vaccines expressing NS1, alone, confer partial protection against ZIKV infection in both immunocompetent and immunodeficient mice, highlighting the value of NS1 as a potential vaccine candidate.
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- 2020
9. A Novel Live Attenuated Respiratory Syncytial Virus Vaccine Candidate with Mutations in the L Protein SAM Binding Site and the G Protein Cleavage Site Is Protective in Cotton Rats and a Rhesus Macaque
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Asuncion Mejias, Michael N. Teng, Christopher J. Walker, Mark E. Peeples, Miaoge Xue, Rongzhang Wang, Phylip Chen, Jianrong Li, Olivia Harder, Octavio Ramilo, Stefan Niewiesk, Tiffany L. Jenkins, and Jacqueline Corry
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Male ,S-Adenosylmethionine ,T cell ,viruses ,Immunology ,Respiratory Syncytial Virus Infections ,Virus Replication ,Microbiology ,Virus ,03 medical and health sciences ,Viral Proteins ,Immune system ,Viral Envelope Proteins ,Virology ,Vaccines and Antiviral Agents ,medicine ,Respiratory Syncytial Virus Vaccines ,Animals ,Humans ,Cotton rat ,Sigmodontinae ,030304 developmental biology ,0303 health sciences ,Attenuated vaccine ,Binding Sites ,biology ,030306 microbiology ,Vaccination ,biology.organism_classification ,Macaca mulatta ,medicine.anatomical_structure ,Insect Science ,Respiratory Syncytial Virus, Human ,Mutation ,biology.protein ,Vero cell ,Respiratory epithelium ,Female ,Antibody - Abstract
Respiratory syncytial virus (RSV) is the leading cause of acute lower respiratory tract infections in children of
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- 2020
10. A Zika virus vaccine expressing premembrane-envelope-NS1 polyprotein
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Mark E. Peeples, Jianrong Li, Prosper N. Boyaka, Ryan N. Jennings, Jingyou Yu, Miaoge Xue, Yuanmei Ma, Zayed Attia, Pei Yong Shi, Mijia Lu, Anzhong Li, Xueya Liang, Nirajkumar Makadiya, Chao Shan, Kelsey Craig, Shan-Lu Liu, and Jennifer J. He
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0301 basic medicine ,Male ,viruses ,General Physics and Astronomy ,Viral Nonstructural Proteins ,Antibodies, Viral ,Zika virus ,Mice ,0302 clinical medicine ,Viral Envelope Proteins ,Vaccines, DNA ,030212 general & internal medicine ,lcsh:Science ,Antigens, Viral ,Mice, Inbred BALB C ,Vaccines, Synthetic ,Multidisciplinary ,biology ,Zika Virus Infection ,Vaccination ,virus diseases ,humanities ,3. Good health ,medicine.anatomical_structure ,Vesicular stomatitis virus ,Cytokines ,Female ,Antibody ,T cell ,Science ,Genetic Vectors ,Vaccines, Attenuated ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,Immune system ,Th2 Cells ,Antigen ,medicine ,Animals ,Polyproteins ,Viral Vaccines ,General Chemistry ,Vesiculovirus ,Zika Virus ,biochemical phenomena, metabolism, and nutrition ,biology.organism_classification ,Virology ,Antibodies, Neutralizing ,Disease Models, Animal ,030104 developmental biology ,Immunization ,biology.protein ,Th17 Cells ,lcsh:Q - Abstract
Current efforts to develop Zika virus (ZIKV) subunit vaccines have been focused on pre-membrane (prM) and envelope (E) proteins, but the role of NS1 in ZIKV-specific immune response and protection is poorly understood. Here, we develop an attenuated recombinant vesicular stomatitis virus (rVSV)-based vaccine expressing ZIKV prM-E-NS1 as a polyprotein. This vectored vaccine candidate is attenuated in mice, where a single immunization induces ZIKV-specific antibody and T cell immune responses that provide protection against ZIKV challenge. Co-expression of prM, E, and NS1 induces significantly higher levels of Th2 and Th17 cytokine responses than prM-E. In addition, NS1 alone is capable of conferring partial protection against ZIKV infection in mice even though it does not induce neutralizing antibodies. These results demonstrate that attenuated rVSV co-expressing prM, E, and NS1 is a promising vaccine candidate for protection against ZIKV infection and highlights an important role for NS1 in ZIKV-specific cellular immune responses., Current Zika virus (ZIKV) subunit vaccine development largely focuses on prM and E proteins, and the role of NS1 for immune response and protection is unclear. Here, Li et al. develop an attenuated VSV-based vaccine expressing a ZIKV prM-E-NS1 polyprotein and characterize immune response and protection in mice.
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- 2018
11. Viral N6-methyladenosine upregulates replication and pathogenesis of human respiratory syncytial virus
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Mark E. Peeples, Zhike Lu, Stefan Niewiesk, Yunsheng Xu, Mijia Lu, Olivia Harder, Chuan He, Jianrong Li, Jiyong Zhou, Phylip Chen, Anzhong Li, Miaoge Xue, Boxuan Simen Zhao, Yuanmei Ma, Xueya Liang, and Zijie Zhang
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Male ,0301 basic medicine ,Adenosine ,Methyltransferase ,Live attenuated vaccines ,Science ,viruses ,General Physics and Astronomy ,Respiratory Syncytial Virus Infections ,Biology ,Virus-host interactions ,Antibodies, Viral ,Vaccines, Attenuated ,Virus Replication ,Article ,General Biochemistry, Genetics and Molecular Biology ,Virus ,03 medical and health sciences ,0302 clinical medicine ,Chlorocebus aethiops ,Gene expression ,Respiratory Syncytial Virus Vaccines ,Animals ,Humans ,Sigmodontinae ,lcsh:Science ,Vero Cells ,Gene knockdown ,Multidisciplinary ,Attenuated vaccine ,Virulence ,General Chemistry ,Methylation ,Virology ,Up-Regulation ,3. Good health ,030104 developmental biology ,Viral replication ,Viral infection ,A549 Cells ,Respiratory Syncytial Virus, Human ,030220 oncology & carcinogenesis ,Vero cell ,RNA ,lcsh:Q ,Female ,HeLa Cells - Abstract
N6-methyladenosine (m6A) is the most prevalent internal modification of mRNAs in most eukaryotes. Here we show that RNAs of human respiratory syncytial virus (RSV) are modified by m6A within discreet regions and that these modifications enhance viral replication and pathogenesis. Knockdown of m6A methyltransferases decreases RSV replication and gene expression whereas knockdown of m6A demethylases has the opposite effect. The G gene transcript contains the most m6A modifications. Recombinant RSV variants expressing G transcripts that lack particular clusters of m6A display reduced replication in A549 cells, primary well differentiated human airway epithelial cultures, and respiratory tracts of cotton rats. One of the m6A-deficient variants is highly attenuated yet retains high immunogenicity in cotton rats. Collectively, our results demonstrate that viral m6A methylation upregulates RSV replication and pathogenesis and identify viral m6A methylation as a target for rational design of live attenuated vaccine candidates for RSV and perhaps other pneumoviruses., Here, Xue et al. identify N6-methyladenosine (m6A) modification sites in RNAs of respiratory syncytial virus (RSV) and show that these sites, particularly sites in the transcript encoding for the viral glycoprotein, affect virus replication in primary human cells and cotton rats.
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- 2019
12. Expression and characterization of a novel truncated rotavirus VP4 for the development of a recombinant rotavirus vaccine
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Shengxiang Ge, Miaoge Xue, Han Yang, Guoxing Luo, Tingdong Li, Lianzhi Jia, Yijian Li, Linqi Yu, Ningshao Xia, and Yuanjun Zeng
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Models, Molecular ,Rotavirus ,0301 basic medicine ,Protein Conformation ,viruses ,medicine.medical_treatment ,Gene Expression ,Antibodies, Viral ,medicine.disease_cause ,law.invention ,Mice ,Immunogenicity, Vaccine ,fluids and secretions ,law ,Gene Order ,Sequence Deletion ,Vaccines, Synthetic ,biology ,Chemistry ,Immunogenicity ,virus diseases ,Rotavirus vaccine ,Recombinant Proteins ,Virus Shedding ,Diarrhea ,Infectious Diseases ,Recombinant DNA ,Molecular Medicine ,Female ,Antibody ,medicine.symptom ,Adjuvant ,Genetic Vectors ,complex mixtures ,Rotavirus Infections ,Cell Line ,03 medical and health sciences ,Neutralization Tests ,medicine ,Animals ,Viral shedding ,General Veterinary ,General Immunology and Microbiology ,Rotavirus Vaccines ,Public Health, Environmental and Occupational Health ,Antibodies, Neutralizing ,Virology ,030104 developmental biology ,biology.protein ,Capsid Proteins ,Immunization - Abstract
The outer capsid protein VP4 is an important target for the development of a recombinant rotavirus vaccine because it mediates the attachment and penetration of rotavirus. Due to the poor solubility of full-length VP4, VP8 was explored as candidate rotavirus vaccines in the past years. In previous studies, it has been found that the N-terminal truncated VP8 protein, VP8-1 (aa26-231), could be expressed in soluble form with improved immunogenicity compared to the core of VP8 (aa65-223). However, this protein stimulated only a weak immune response when aluminum hydroxide was used as an adjuvant. In addition, it should be noted that the protective efficacy of VP4 was higher than that of VP8 and VP5. In this study, it was found that when the N-terminal 25 amino acids were deleted, the truncated VP4∗ (aa26-476) containing VP8 and the stalk domain of VP5 could be expressed in soluble form in E. coli and purified to homogeneous trimers. Furthermore, the truncated VP4 could induce high titers of neutralizing antibodies when aluminum adjuvant was used and conferred high protective efficacy in reducing the severity of diarrhea and rotavirus shedding in stools in animal models. The immunogenicity of the truncated VP4 was significantly higher than that of VP8∗ and VP5∗ alone. Taken together, the truncated VP4∗ (aa26-476), with enhanced immunogenicity and immunoprotectivity, could be considered as a viable candidate for further development and has the potential to become a parenterally administered rotavirus vaccine.
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- 2018
13. A Lactic Acid Bacteria (LAB)-Based Vaccine Candidate for Human Norovirus
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Thomas Z. Gao, Xianjun Dai, Miaoge Xue, Ryan N. Jennings, Andrew J. Niehaus, Lucia E. Rosas, Jianrong Li, Anzhong Li, Kelsey Craig, and Mijia Lu
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0301 basic medicine ,DNA, Bacterial ,human norovirus ,Swine ,viruses ,030106 microbiology ,Genetic Vectors ,lcsh:QR1-502 ,Ileum ,medicine.disease_cause ,Antibodies, Viral ,lcsh:Microbiology ,Virus ,Article ,Microbiology ,03 medical and health sciences ,Feces ,Antigen ,Virology ,vaccine ,medicine ,gnotobiotic piglet ,Mesenteric lymph nodes ,Animals ,Germ-Free Life ,Caliciviridae Infections ,Viral Structural Proteins ,biology ,Lactococcus lactis ,Norovirus ,Viral Vaccines ,VP1 ,biology.organism_classification ,Small intestine ,Gastroenteritis ,lactic acid bacteria ,Gastrointestinal Tract ,030104 developmental biology ,Infectious Diseases ,medicine.anatomical_structure - Abstract
Human noroviruses (HuNoVs) are responsible for more than 95% of the non-bacterial acute gastroenteritis epidemics in the world. The CDC estimates that every year 21 million individuals suffer from HuNoV-induced gastroenteritis in the United States. Currently, there is no FDA-approved vaccine for HuNoVs. Development of an effective vaccine has been hampered by the lack of an efficient cell culture system for HuNoVs and a suitable small animal model for pathogenesis study. In this study, we developed lactic acid bacteria (LAB) as a vector to deliver HuNoV antigen. A LAB strain (Lactococcus lactis) carrying VP1 gene of a HuNoV GII.4 virus (LAB-VP1) was constructed. It was found that HuNoV VP1 protein was highly expressed by LAB vector and was secreted into media supernatants. To test whether LAB-based HuNoV vaccine candidate is immunogenic, 4-day-old gnotobiotic piglets were orally inoculated with various doses of LAB-VP1. It was found that LABs were persistent in the small intestine of piglets and shed in pig feces for at least 25 days post inoculation. LAB DNA and VP1 were detected in mesenteric lymph nodes and spleen tissue in LAB-VP1 inoculated groups. HuNoV-specific IgG and IgA were detectable in serum and feces respectively at day 13 post-inoculation, and further increased at later time points. After being challenged with HuNoV GII.4 strain, a large amount of HuNoV antigens were observed in the duodenum, jejunum, and ileum sections of the intestine in the LAB control group. In contrast, significantly less, or no, HuNoV antigens were detected in the LAB-VP1 immunized groups. Collectively, these results demonstrate that a LAB-based HuNoV vaccine induces protective immunity in gnotobiotic piglets.
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- 2019
14. N
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Mijia, Lu, Zijie, Zhang, Miaoge, Xue, Boxuan Simen, Zhao, Olivia, Harder, Anzhong, Li, Xueya, Liang, Thomas Z, Gao, Yunsheng, Xu, Jiyong, Zhou, Zongdi, Feng, Stefan, Niewiesk, Mark E, Peeples, Chuan, He, and Jianrong, Li
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Adenosine ,Interferon-Induced Helicase, IFIH1 ,THP-1 Cells ,viruses ,Genome, Viral ,Article ,Chlorocebus aethiops ,Animals ,Humans ,Sigmodontinae ,Receptors, Immunologic ,Vero Cells ,Immune Evasion ,Paramyxoviridae Infections ,NF-kappa B ,Virion ,virus diseases ,Interferon-beta ,Gene Expression Regulation ,A549 Cells ,Host-Pathogen Interactions ,DEAD Box Protein 58 ,RNA, Viral ,Interferon Regulatory Factor-3 ,Metapneumovirus ,HeLa Cells ,Signal Transduction - Abstract
Internal N6-methyladenosine (m6A) modification is one of the most common and abundant modifications of RNA. However, the biological role(s) of viral RNA m6A remains elusive. Using human metapneumovirus (hMPV) as a model, we demonstrate that m6A serves as a molecular marker for innate immune discrimination of self from nonself RNAs. We show that hMPV RNAs are m6A methylated and that viral m6A methylation promotes hMPV replication and gene expression. Inactivating m6A addition sites with synonymous mutations or demethylase resulted in m6A deficient recombinant hMPVs and virion RNAs that induced significantly higher expression of type I interferon (IFN) which was dependent on the cytoplasmic RNA sensor RIG-I, not MDA5. Mechanistically, m6A-deficient virion RNA induces higher expression of RIG-I, binds more efficiently to RIG-I, and facilitates the conformational change of RIG-I, leading to enhanced IFN expression. Furthermore, m6A-deficient rhMPVs triggered higher IFN in vivo and were significantly attenuated in cotton rats yet retained high immunogenicity. Collectively, our results highlight that (i) virus acquires m6A in their RNAs as a means of mimicking cellular RNA to avoid detection by innate immunity; and (ii) viral RNA m6A can serve as a target to attenuate hMPV for vaccine purposes.
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- 2019
15. Immunogenicity and protective efficacy of rotavirus VP8*fused to cholera toxin B subunit in a mouse model
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Shengxiang Ge, Yuanjun Zeng, Lianzhi Jia, Ningshao Xia, Yijian Li, Miaoge Xue, Tingdong Li, and Linqi Yu
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0301 basic medicine ,Cholera Toxin ,medicine.drug_class ,Recombinant Fusion Proteins ,medicine.medical_treatment ,Immunology ,Viral Nonstructural Proteins ,Antibodies, Viral ,medicine.disease_cause ,Monoclonal antibody ,complex mixtures ,Rotavirus Infections ,03 medical and health sciences ,0302 clinical medicine ,Adjuvants, Immunologic ,Rotavirus ,medicine ,Animals ,Immunology and Allergy ,030212 general & internal medicine ,Neutralizing antibody ,Pharmacology ,Mice, Inbred BALB C ,Vaccines, Synthetic ,biology ,Chemistry ,Immunogenicity ,Cholera toxin ,Rotavirus Vaccines ,RNA-Binding Proteins ,Antibodies, Neutralizing ,Research Papers ,Virology ,Fusion protein ,Disease Models, Animal ,030104 developmental biology ,Immunization ,Vaccines, Subunit ,biology.protein ,Adjuvant - Abstract
In attempts to develop recombinant subunit vaccines against rotavirus disease, it was previously shown that the N-terminal truncated VP8* protein, VP8-1 (aa26-231), is a good vaccine candidate when used for immunization in combination with Freund's adjuvant. However, this protein stimulated only weak immune response when aluminum hydroxide was used as an adjuvant. In this study, the nontoxic B subunit of cholera toxin (CTB) was employed as intra-molecular adjuvant to improve the immunogenicity of VP8-1. Both, the N-terminal and C-terminal fusion proteins, were purified to homogeneity, at which stage they formed pentamers, and showed significantly higher immunogenicity and protective efficacy than a VP8-1/aluminum hydroxide mixture in a mouse model. Compared to VP8-1-CTB, CTB-VP8-1 showed higher binding activity to both, GM1 and the conformation sensitive neutralizing monoclonal antibodies specific to VP8. More importantly, CTB-VP8-1 elicited higher titers of neutralizing antibodies and conferred higher protective efficacy than VP8-1-CTB. Therefore, the protein CTB-VP8-1, with enhanced immunogenicity and immunoprotectivity, could be considered as a viable candidate for further development of an alternative, replication-incompetent, parenterally administered vaccine against rotavirus disease.
- Published
- 2016
16. Nonsegmented Negative-Sense RNA Viruses Utilize N6-Methyladenosine (m6A) as a Common Strategy To Evade Host Innate Immunity.
- Author
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Mijia Lu, Miaoge Xue, Hai-Tao Wang, Kairis, Elizabeth L., Ahmad, Sadeem, Jiangbo Wei, Zijie Zhang, Qinzhe Liu, Yuexiu Zhang, Youling Gao, Garcin, Dominique, Peeples, Mark E., Sharma, Amit, Sun Hur, Chuan He, and Jianrong Li
- Subjects
- *
NATURAL immunity , *TYPE I interferons , *RNA viruses , *RNA methylation , *RNA modification & restriction , *PENTRAXINS , *CARRIER proteins - Abstract
N6-Methyladenosine (m6A) is the most abundant internal RNA modification catalyzed by host RNA methyltransferases. As obligate intracellular parasites, many viruses acquire m6A methylation in their RNAs. However, the biological functions of viral m6A methylation are poorly understood. Here, we found that viral m6A methylation serves as a molecular marker for host innate immunity to discriminate self from nonself RNA and that this novel biological function of viral m6A methylation is universally conserved in several families in nonsegmented negative-sense (NNS) RNA viruses. Using m6A methyltransferase (METTL3) knockout cells, we produced m6A-deficient virion RNAs from the representative members of the families Pneumoviridae, Paramyxoviridae, and Rhabdoviridae and found that these m6A-deficient viral RNAs triggered significantly higher levels of type I interferon compared to the m6A-sufficient viral RNAs, in a RIG-I-dependent manner. Reconstitution of the RIG-I pathway revealed that m6A-deficient virion RNA induced higher expression of RIG-I, bound to RIG-I more efficiently, enhanced RIG-I ubiquitination, and facilitated RIG-I conformational rearrangement and oligomerization. Furthermore, the m6A binding protein YTHDF2 is essential for suppression of the type I interferon signaling pathway, including by virion RNA. Collectively, our results suggest that several families in NNS RNA viruses acquire m6A in viral RNA as a common strategy to evade host innate immunity. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
17. Internal N6-methyladenosine modification controls the innate immune response to viral RNA
- Author
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miaoge xue, Mijia Lu, Anzhong Li, Zijie Zhang, Chuan He, and Jianrong Li
- Subjects
Immunology ,Immunology and Allergy - Abstract
Internal N6-methyladenosine (m6A) modification is one of the most prevalent and abundant modifications of RNA. The m6A methylation is catalyzed by host methyltransferases (METTL3 and METTL14) and can be reversibly removed by host demethylases (FTO and ALKBH5). Recent studies have revealed that RNA of many DNA and RNA viruses contain m6A methylation and that viral m6A can play a pro-viral or anti-viral role. However, the biological functions of viral RNA m6A methylation remain poorly understood. In this study, we found that genome and antigenome of human respiratory syncytial virus (RSV), a non-segmented negative-sense RNA virus, are m6A methylated and that viral m6A methylation positively regulates RSV replication and gene expression. Interestingly, virion RNAs purified from RSV grown in METTL3-knockout cells induced significantly higher type I interferon responses than virion RNAs purified from RSV grown in wild type cells, demonstrating that viral RNA m6A methylation inhibits innate immunity. Similarly, inactivation of m6A methylation in RSV genome and/or antigenome by site-directed mutagenesis induced significantly higher innate immune responses than wild type RSV RNA. Mechanistically, these m6A-unmodified or m6A-deficient RSV RNAs trigger a higher expression of pattern recognition receptors (such as RIG-I and MDA5), enhance their binding affinity to the innate immune RNA sensors, and enhance the phosphorylation of downstream transcription factor IRF3, leading to an enhanced activation of type I interferon signaling pathway. Taken together, our study identified a novel biological function of viral m6A methylation, which is to serves as a molecular marker for host innate immunity to discriminate self from nonself RNAs.
- Published
- 2020
18. Characterization and protective efficacy in an animal model of a novel truncated rotavirus VP8 subunit parenteral vaccine candidate
- Author
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Shengxiang Ge, Miaoge Xue, Yuanjun Zeng, Ningshao Xia, Tingdong Li, Linqi Yu, Yaojian Che, Haijun Lin, and Mujin Fang
- Subjects
Diarrhea ,Rotavirus ,Injections, Subcutaneous ,Protein subunit ,Viral Nonstructural Proteins ,Biology ,Antibodies, Viral ,medicine.disease_cause ,Rotavirus Infections ,Escherichia coli ,medicine ,Animals ,Neutralizing antibody ,Mice, Inbred BALB C ,Vaccines, Synthetic ,General Veterinary ,General Immunology and Microbiology ,Immunogenicity ,Rotavirus Vaccines ,Public Health, Environmental and Occupational Health ,RNA-Binding Proteins ,Antibodies, Neutralizing ,Virology ,Disease Models, Animal ,Titer ,Infectious Diseases ,Immunization ,Vaccines, Subunit ,biology.protein ,Molecular Medicine ,Female ,medicine.symptom ,Immunity, Maternally-Acquired - Abstract
The cell-attachment protein VP8* of rotavirus is a potential candidate parenteral vaccine. However, the yield of full-length VP8 protein (VP8*, residues 1-231) expressed in Escherichia coli was low, and a truncated VP8 protein (ΔVP8*, residues 65-231) cannot elicit efficient protective immunity in a mouse model. In this study, tow novel truncated VP8 proteins, VP8-1 (residues 26-231) and VP8-2 (residues 51-231), were expressed in E. coli and evaluated for immunogenicity and protective efficacy, compared with VP8* and ΔVP8*. As well as ΔVP8*, the protein VP8-1 and VP8-2 were successfully expressed in high yield and purified in homogeneous dimeric forms, while the protein VP8* was expressed with lower yield and prone to aggregation and degradation in solution. Although the immunogenicity of the protein VP8*, VP8-1, VP8-2 and ΔVP8* was comparable, immunization of VP8* and VP8-1 elicited significantly higher neutralizing antibody titers than that of VP8-2 and ΔVP8* in mice. Furthermore, when assessed using a mouse maternal antibody model, the efficacy of VP8-1 to protect against rotavirus-induced diarrhea in pups was comparable to that of VP8*, both were dramatically higher than that of VP8-2 and ΔVP8*. Taken together, the novel truncated protein VP8-1, with increased yield, improved homogeneity and high protective efficacy, is a viable candidate for further development of a parenterally administrated prophylactic vaccine against rotavirus infection.
- Published
- 2015
19. Development of an enzyme-linked immunospot assay for determination of rotavirus infectivity
- Author
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Qinjian Zhao, Ningshao Xia, Shengxiang Ge, Tingdong Li, Jun Zhang, Haijun Lin, Miaoge Xue, and Linqi Yu
- Subjects
Rotavirus ,Enzyme-Linked Immunospot Assay ,medicine.drug_class ,viruses ,Biology ,Antibodies, Viral ,Monoclonal antibody ,medicine.disease_cause ,Neutralization ,Cell Line ,Inhibitory Concentration 50 ,Neutralization Tests ,Virology ,Chlorocebus aethiops ,Genotype ,medicine ,Animals ,IC50 ,Horseradish Peroxidase ,Infectivity ,Staining and Labeling ,ELISPOT ,virus diseases ,Viral Load ,Titer - Abstract
Conventional rotavirus infectivity assays are time consuming, labor intensive, and with low sample throughput. To overcome these problems, a 96-well microplate enzyme-linked immunospot assay (Elispot) was developed for the measurement of rotavirus infectious titers. The infected MA104 cells were stained with a horseradish peroxidase-conjugated anti-VP6 monoclonal antibody followed by detection with an ELISPOT analyzer. A linear relationship was found between spot number and input of rotavirus dose in SA11 and 10 rotavirus isolates of different genotypes. The propagation of rotavirus SA11 in MA104 cells was monitored, and the neutralizing activity of serum samples and monoclonal antibodies was determined. The 50% neutralizing titer (NT50) of serum and 50% inhibitory concentration (IC50) of monoclonal antibodies were correlated well with the results determined by ELISA-based neutralization assay. In conclusion, a rapid and semi-automated procedure to determine rotavirus infectivity was developed, which will be useful to study the infectivity and the neutralizing epitopes of rotavirus.
- Published
- 2014
20. Stable Attenuation of Human Respiratory Syncytial Virus for Live Vaccines by Deletion and Insertion of Amino Acids in the Hinge Region between the mRNA Capping and Methyltransferase Domains of the Large Polymerase Protein.
- Author
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Miaoge Xue, Rongzhang Wang, Harder, Olivia, Chen, Phylip, Mijia Lu, Hui Cai, Anzhong Li, Xueya Liang, Jennings, Ryan, La Perle, Krista, Niewiesk, Stefan, Peeples, Mark E., and Jianrong Li
- Subjects
- *
RESPIRATORY syncytial virus , *VIRAL vaccines , *MESSENGER RNA , *AMINO acids , *RNA replicase , *METHYLTRANSFERASES , *RNA polymerases - Abstract
Human respiratory syncytial virus (RSV) is the leading viral cause of lower respiratory tract disease in infants and children worldwide. Currently, there are no FDA-approved vaccines to combat this virus. The large (L) polymerase protein of RSV replicates the viral genome and transcribes viral mRNAs. The L protein is organized as a core ring-like domain containing the RNA-dependent RNA polymerase and an appendage of globular domains containing an mRNA capping region and a cap methyltransferase region, which are linked by a flexible hinge region. Here, we found that the flexible hinge region of RSV L protein is tolerant to amino acid deletion or insertion. Recombinant RSVs carrying a single or double deletion or a single alanine insertion were genetically stable, highly attenuated in immortalized cells, had defects in replication and spread, and had a delay in innate immune cytokine responses in primary, well-differentiated, human bronchial epithelial (HBE) cultures. The replication of these recombinant viruses was highly attenuated in the upper and lower respiratory tracts of cotton rats. Importantly, these recombinant viruses elicited high levels of neutralizing antibody and provided complete protection against RSV replication. Taken together, amino acid deletions or insertions in the hinge region of the L protein can serve as a novel approach to rationally design genetically stable, highly attenuated, and immunogenic live virus vaccine candidates for RSV. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
21. A Convenient Nucleic Acid Test on the Basis of the Capillary Convective PCR for the On-Site Detection of Enterovirus 71
- Author
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Ping-Hei Chen, Pingguo Liu, Ningshao Xia, Shengxiang Ge, Jieyu Chen, Penglin Wang, Jin Wang, Shuizhen He, Yanyan Lin, Xu Feihai, Zhou Wenbin, Miaoge Xue, and Shiyin Zhang
- Subjects
China ,Pcr cloning ,Diagnosis tool ,Biology ,Polymerase Chain Reaction ,Sensitivity and Specificity ,Pathology and Forensic Medicine ,law.invention ,law ,medicine ,Enterovirus 71 ,Humans ,Child ,Polymerase chain reaction ,medicine.diagnostic_test ,Nucleic acid test ,Equipment Design ,Dipstick ,biology.organism_classification ,Virology ,Enterovirus A, Human ,Visual detection ,Nucleic acid ,RNA, Viral ,Molecular Medicine ,Hand, Foot and Mouth Disease - Abstract
The recent and continuing epidemic of enterovirus 71 in China has affected millions of children and resulted in thousands of deaths. Timely diagnosis and management is essential for disease control. Current enterovirus 71 molecular tests require resources that are unavailable for on-site testing. We have developed a simple-to-operate nucleic acid test, the convenient and integrated nucleic acid test, for local medical institutions. It uses a convective PCR for rapid amplification, a dipstick for visual detection of PCR products, and a simple commercial kit for nucleic acid extraction. By using a specially designed reagent and reaction tube containing a dipstick, the amplification and detection processes are well integrated and simplified. Moreover, cross contamination that may be caused by an open-tube detection system can be avoided. On the basis of the convenient and integrated nucleic acid test, an enterovirus 71 assay for on-site testing was developed. After evaluating known hand, foot, and mouth disease virus stocks of 17 strains of 11 different serotypes, this assay showed a favorable detection spectrum and no cross-reactivity. Its clinical performance was established by testing 141 clinical samples and comparing the results with a nested RT-PCR method. The assay showed a clinical sensitivity and specificity of 98.5% and 100%, respectively. Our results suggest that this convenient and integrated nucleic acid test enterovirus 71 assay may serve as an on-site diagnosis tool.
- Published
- 2014
22. A one-step dipstick assay for the on-site detection of nucleic acid
- Author
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Zhou Wenbin, Shengxiang Ge, Jing Zhang, Ningshao Xia, Jieyu Chen, Shiyin Zhang, Zhantong Wang, Qingrui Chen, Ping-Hei Chen, and Miaoge Xue
- Subjects
Hepatitis B virus ,Engineering ,Technology research ,business.industry ,Melting temperature ,Clinical Biochemistry ,Human immunodeficiency virus (HIV) ,Hepacivirus ,General Medicine ,Dipstick ,medicine.disease_cause ,Dna amplification ,Polymerase Chain Reaction ,Sensitivity and Specificity ,Biochemistry ,DNA, Viral ,HIV-1 ,Nucleic acid ,medicine ,Humans ,Human papillomavirus ,business ,Software engineering ,Nucleic Acid Amplification Techniques - Abstract
We have developed a one-step nucleic acid dipstick assay (NADA) for visually detecting polymerase chain reaction (PCR) products within 3min. "One-step" means that there were no additional procedures between amplification and detection.This method was achieved through the use of asymmetric PCR and specially designed probes with appropriate melting temperature values. We initially combined one-step NADA with asymmetric capillary convective PCR (ACCPCR), an easy and rapid nucleic acid amplification technique, to construct an on-site nucleic acid diagnostic platform.We developed a diagnostic assay for the hepatitis B virus based on the ACCPCR-NADA platform to verify its feasibility. It exhibited an analytical sensitivity of three copies per test and a broad detection spectrum including genotype A-I. It also showed 97.9% sensitivity and 100% specificity based on the results observed using 67 serum samples with the Roche COBAS AmpliPrep/COBAS TaqMan (COBAS) system as the standard for comparison.The results provide evidence for the feasibility of using an ACCPCR-NADA platform in practical applications, especially in on-site test.
- Published
- 2013
23. Vesicular Stomatitis Virus and DNA Vaccines Expressing Zika Virus Nonstructural Protein 1 Induce Substantial but Not Sterilizing Protection against Zika Virus Infection.
- Author
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Anzhong Li, Miaoge Xue, Zayed Attia, Jingyou Yu, Mijia Lu, Chao Shan, Xueya Liang, Gao, Thomas Z., Pei-Yong Shi, Peeples, Mark E., Boyaka, Prosper N., Shan-Lu Liu, and Jianrong Li
- Subjects
- *
ZIKA virus infections , *VIRAL nonstructural proteins , *VESICULAR stomatitis , *DNA vaccines , *ZIKA virus , *VIRAL vaccines , *ARBOVIRUS diseases - Abstract
The nonstructural protein 1 (NS1) of several flaviviruses, including West Nile, dengue, and yellow fever viruses, is capable of inducing variable degrees of protection against flavivirus infection in animal models. However, the immunogenicity of NS1 protein of Zika virus (ZIKV) is less understood. Here, we determined the efficacy of ZIKV NS1-based vaccine candidates using two delivery platforms, methyltransferase-defective recombinant vesicular stomatitis virus (mtdVSV) and a DNA vaccine. We first show that expression of ZIKV NS1 could be significantly enhanced by optimizing the signal peptide. A single dose of mtdVSV-NS1-based vaccine or two doses of DNA vaccine induced high levels of NS1-specfic antibody and T cell immune responses but provided only partial protection against ZIKV viremia in BALB/c mice. In Ifnar1-/- mice, neither NS1-based vaccine provided protection against a lethal high dose (105 PFU) ZIKV challenge, but mtdVSV-NS1-based vaccine prevented deaths from a low dose (10³ PFU) challenge, though they experienced viremia and body weight loss. We conclude that ZIKV NS1 alone conferred substantial, but not complete, protection against ZIKV infection. Nevertheless, these results highlight the value of ZIKV NS1 for vaccine development. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
24. A Novel Live Attenuated Respiratory Syncytial Virus Vaccine Candidate with Mutations in the L Protein SAM Binding Site and the G Protein Cleavage Site Is Protective in Cotton Rats and a Rhesus Macaque.
- Author
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Jenkins, Tiffany, Rongzhang Wang, Harder, Olivia, Miaoge Xue, Chen, Phylip, Corry, Jacqueline, Walker, Christopher, Teng, Michael, Mejias, Asuncion, Ramilo, Octavio, Niewiesk, Stefan, Jianrong Li, and Peeples, Mark E.
- Subjects
- *
RESPIRATORY syncytial virus , *RHESUS monkeys , *VIRAL vaccines , *G proteins , *PROTEIN binding , *G protein coupled receptors , *HUMAN metapneumovirus infection , *METHYLTRANSFERASES - Abstract
Respiratory syncytial virus (RSV) is the leading cause of acute lower respiratory tract infections in children of >5 years of age worldwide, infecting the majority of infants in their first year of life. Despite the widespread impact of this virus, no vaccine is currently available. For more than 50 years, live attenuated vaccines (LAVs) have been shown to protect against other childhood viral infections, offering the advantage of presenting all viral proteins to the immune system for stimulation of both B and T cell responses and memory. The RSV LAV candidate described here, rgRSV-L(G1857A)-G(L208A), contains two modifications: an attenuating mutation in the S-adenosylmethionine (SAM) binding site of the viral mRNA cap methyltransferase (MTase) within the large (L) polymerase protein and a mutation in the attachment (G) glycoprotein that inhibits its cleavage during production in Vero cells, resulting in virus with a "noncleaved G" (ncG). RSV virions containing the ncG have an increased ability to infect primary well-differentiated human bronchial epithelial (HBE) cultures which model the in vivo site of immunization, the ciliated airway epithelium. This RSV LAV candidate is produced efficiently in Vero cells, is highly attenuated in HBE cultures, efficiently induces neutralizing antibodies that are long lasting, and provides protection against an RSV challenge in the cotton rat, without causing enhanced disease. Similar results were obtained in a rhesus macaque. IMPORTANCE Globally, respiratory syncytial virus (RSV) is a major cause of death in children under 1 year of age, yet no vaccine is available. We have generated a novel RSV live attenuated vaccine candidate containing mutations in the L and G proteins. The L polymerase mutation does not inhibit virus yield in Vero cells, the cell type required for vaccine production, but greatly reduces virus spread in human bronchial epithelial (HBE) cultures, a logical in vitro predictor of in vivo attenuation. The G attachment protein mutation reduces its cleavage in Vero cells, thereby increasing vaccine virus yield, making vaccine production more economical. In cotton rats, this RSV vaccine candidate is highly attenuated at a dose of 105 PFU and completely protective following immunization with 500 PFU, 200-fold less than the dose usually used in such studies. It also induced long-lasting antibodies in cotton rats and protected a rhesus macaque from RSV challenge. This mutant virus is an excellent RSV live attenuated vaccine candidate. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
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