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Multipotent mesenchymal stromal cells are fully permissive for human cytomegalovirus infection

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Virologica Sinica 2016年第3期31卷 219-228页

作者： Guan-hua Qiao Fei Zhao Shuang Cheng min-hua luo State Key Laboratory of Virology Wuhan Institute of VirologyChinese Academy of Sciences

Congenital human cytomegalovirus(HCMV) infection is a leading infectious cause of birth *** studies have reported birth defects with multiple organ maldevelopment in congenital HCMV-infected neonates. Multipotent mesenchymal stromal cells(MSCs) are a group of stem/progenitor cells that are multi-potent and can self-renew, and they play a vital role in multiorgan formation. Whether MSCs are susceptible to HCMV infection is unclear. In this study, MSCs were isolated from Wharton's jelly of the human umbilical cord and identified by their plastic adherence, surface marker pattern, and differentiation capacity. Then, the MSCs were infected with the HCMV Towne strain, and infection status was assessed via determination of viral entry,replication initiation, viral protein expression, and infectious virion release using western blotting,immunofluorescence assays, and plaque forming assays. The results indicate that the isolated MSCs were fully permissive for HCMV infection and provide a preliminary basis for understanding the pathogenesis of HCMV infection in non-nervous system diseases, including multi-organ malformation during fetal development.

关键词： human cytomegalovirus(HCMV) multipotent mesenchymal stromal cells(MSCs) susceptibility umbilical cord Wharton's jelly

Concerns on Vaccine against Varicella Caused by Varicella-Zoster Virus Infection

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Virologica Sinica 2021年第1期36卷 159-162页

作者： Wen-Bo Zeng Fukun Zhang Shuang Cheng Jin-yan Sun Hongjie Shen min-hua luo State Key Laboratory of Virology CAS Center for Excellence in Brain Science and Intelligence Technology(CEBSIT)Center for Biosafety Mega-ScienceWuhan Institute of VirologyChinese Academy of SciencesWuhan 430071China Changchun Keygen Biological Products Co.Ltd Changchun 130000China University of Chinese Academy of Sciences Beijing 100049China

Varicella-zoster virus(VZV)belongs to a neurotropic Alphaherpesvirinae and is the causative pathogen of both the varicella and herpes ***,there are three different vaccines for the prevention,low does live-attenuated Oka(v Oka)against varicella,high does v Oka and glycoprotein E with the AS01B adjuvant system against *** vaccine containing v Oka effectively prevents varicella,but latency of v Oka can be detected in the inoculated population and its reactivation causes zoster.

关键词： zoster prevention vaccine

Anterograde Trans-Synaptic Tagging Mediated by Adeno-Associated Virus

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Neuroscience Bulletin 2017年第3期33卷 348-350页

作者： Fei Zhao Hai-Fei Jiang Wen-Bo Zeng Yousheng Shu min-hua luo Shumin Duan State Key Laboratory of Virology Center for Excellence in Brain Science and Intelligence Technology Chinese Academy of Sciences Wuhan Institute of Virology Chinese Academy of Sciences State Key Laboratory of Cognitive Neuroscience and Learning IDG/Mc Govern Institute for Brain ResearchSchool of Brain and Cognitive Sciences the Collaborative Innovation Center for Brain Science Beijing Normal University Department of Neurobiology Key Laboratory of Medical Neurobiology of the Ministry of Health of China Key Laboratory of Neurobiology Zhejiang University School of Medicine

Trans-neuronal viral tracing is becoming one of the most important methods for mapping neuronal circuit connections,but an anterograde trans-synaptic tracer compatible with functional assays is still ***-associated virus（AAV）is replication-defective,and its production of offspring needs the help of adenovirus（AV）[1]or herpesvirus co-infection[2].The replication deficiency,non-pathogenicity,easy manipulability,

关键词： neuronal herpesvirus offspring replication pathogenicity synaptic tracing Associated adenovirus titer

Human cytomegalovirus infection dysregulates neural progenitor cell fate by disrupting Hes1 rhythm and down-regulating its expression

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Virologica Sinica 2017年第3期32卷 188-198页

作者： Xi-Juan Liu Xuan Jiang Sheng-Nan huang Jin-Yan Sun Fei Zhao Wen-Bo Zeng min-hua luo State Key Laboratory of Virology CAS Center for Excellence in Brain Science and Intelligence Technology (CEBSIT) Wuhan Institute of Virology Chinese Academy of Sciences University of Chinese Academy of Sciences The Joint Center of Translational Precision Medicine Guangzhou Institute of Pediatrics Guangzhou Women and Children Medical Center

Human cytomegalovirus(HCMV) infection is a leading cause of birth defects, primarily affecting the central nervous system and causing its maldevelopment. As the essential downstream effector of Notch signaling pathway, Hes1, and its dynamic expression, plays an essential role on maintaining neural progenitor/stem cells(NPCs) cell fate and fetal brain development. In the present study, we reported the first observation of Hes1 oscillatory expression in human NPCs, with an approximately1.5 hour periodicity and a Hes1 protein half-life of about 17(17.6 ± 0.2) minutes. HCMV infection disrupts the Hes1 rhythm and down-regulates its expression. Furthermore, we discovered that depleting Hes1 protein disturbed NPCs cell fate by suppressing NPCs proliferation and neurosphere formation, and driving NPCs abnormal differentiation. These results suggested a novel mechanism linking disruption of Hes1 rhythm and down-regulation of Hes1 expression to neurodevelopmental disorders caused by congenital HCMV infection.

关键词： human cytomegalovirus(HCMV) neural progenitor cells(NPCs) Hes1 rhythm cell fate

Construction and characterization of a synthesized herpes simplex virus H129-Syn-G2

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Virologica Sinica 2023年第3期38卷 373-379页

作者： Han Xiao Hengrui Hu Yijia Guo Jiang Li Le Wen Wen-Bo Zeng Manli Wang min-hua luo Zhihong Hu State Key Laboratory of Virology Center for Biosafety Mega-ScienceWuhan Institute of VirologyChinese Academy of SciencesWuhan430071China University of Chinese Academy of Sciences Beijing100049China

Herpes simplex virus type 1(HSV-1)causes lifelong infections worldwide,and currently there is no efficient cure or ***-1-derived tools,such as neuronal circuit tracers and oncolytic viruses,have been used exten-sively;however,further genetic engineering of HSV-1 is hindered by its complex genome *** the present study,we designed and constructed a synthetic platform for HSV-1 based on *** complete genome was constructed from 10 fragments through 3 rounds of synthesis using transformation-associated recombination(TAR)in yeast,and was named *** H129-Syn-G2 genome contained two copies of the gfp gene and was transfected into cells to rescue the *** to growth curve assay and electron microscopy results,the synthetic viruses exhibited more optimized growth properties and similar morphogenesis compared to the parental *** synthetic platform will facilitate further manipulation of the HSV-1 genome for the devel-opment of neuronal circuit tracers,oncolytic viruses,and vaccines.

关键词： Herpes simplex virus type 1(HSV-1) Neuronal circuit tracers H129-Syn-G2 H129-G4 Synthetic biology

The Susceptibility of Primary Dermis Fibroblasts from the Chinese Tree Shrew to Human Cytomegalovirus Infection

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Virologica Sinica 2019年第3期34卷 270-277页

作者： Shu-Wei Dong Ling-Shuai Jiao ming Yang Ying-Liang Duan Yi-Bo Chen Fei Zhao A-Mei Zhang Li Liu min-hua luo Xue-Shan Xia Faculty of Life Science and Technology Kunming University of Science and TechnologyKunming 650500China The Key Laboratory of Tropical and Subtropical Animal Viral Diseases in Yunnan Province Kunming 650224China State Key Laboratory of Virology Wuhan Institute of VirologyChinese Academy of SciencesWuhan 430071China

As a universal pathogen leading to neonatal defects and transplant failure,human cytomegalovirus(HCMV)has strict species specificity and this has prevented the development of a suitable animal model for the pathogenesis *** mechanism of cross-species barrier remains elusive and there are so far no non-human cell culture models that support HCMV *** Chinese tree shrew(Tupaia belangeri chinensis)is a small laboratory animal and evolutionary closely related with *** investigated the susceptibility of primary tree shrew dermis fibroblasts(TSDF)to HCMV *** with a GFP-expressing HCMV virus resulted in green fluorescence in infected cells with the expression of IE1,UL44 and *** titers of cell-free viruses reached 10^3 PFU/mL at 96 hpi,compared to titers of 10^4 PFU/mL observed in primary human foreskin *** results suggested that TSDF was semi-permissive for HCMV *** TSDF model could be further used to investigate key factors influencing cross-species multiplication of HCMV.

关键词： Human cytomegalovirus(HCMV) Primary tree shrew dermis fibroblasts(TSDF) Cross-species infection Semi-permissiveness

HSV-1 H129-Derived Anterograde Neural Circuit Tracers:Improvements, Production, and Applications

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Neuroscience Bulletin 2021年第5期37卷 701-719页

作者： Hong Yang Feng Xiong Yi-Ge Song Hai-Fei Jiang Hai-Bin Qin Jing Zhou Sha Lu Steven FGrieco Xiangmin Xu Wen-Bo Zeng Fei Zhao min-hua luo State Key Laboratory of Virology CAS Center for Excellence in Brain Science and Intelligence TechnologyCenter for Biosafety Mega-ScienceWuhan Institute of VirologyChinese Academy of SciencesWuhan 430071China University of Chinese Academy of Sciences Beijing 100049China Shanghai Genechem Co.Ltd.Shanghai 201203 China Department of Anatomy and Neurobiology School of MedicineUniversity of CaliforniaIrvineCA 92697USA School of Basic Medical Sciences Capital Medical UniversityBeijing 100069China Chinese Institute for Brain Research Beijing 102206China Shanghai Public Health Clinical Center Fudan UniversityShanghai 200032China

Anterograde viral tracers are powerful and essential tools for dissecting the output targets of a brain region of interest. They have been developed from herpes simplex virus 1(HSV-1) strain H129(H129), and have been successfully applied to map diverse neural ***, the anterograde polysynaptic tracer H129-G4 was used by many groups. We then developed the first monosynaptic tracer, H129-dTK-tdT, which was highly successful, yet improvements are needed. Now, by inserting another tdTomato expression cassette into the H129-dTK-tdT genome, we have created H129-dTK-T2, an updated version of H129-dTK-tdT that has improved labeling intensity. To help scientists produce and apply our H129-derived viral tracers, here we provide the protocol describing our detailed and standardized procedures. Commonly-encountered technical problems and their solutions are also discussed in detail. Broadly, the dissemination of this protocol will greatly support scientists to apply these viral tracers on a large scale.

关键词： HSV-1 strain H129(H129) Anterograde transneuronal tracer Polysynaptic tracer Monosynaptic tracer Production Application Neural circuit Neural circuit tracing

Human cytomegalovirus DNA and immediate early protein 1/2 are highly associated with glioma and prognosis

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Protein & Cell 2020年第7期11卷 525-533页

作者： Le Wen Fei Zhao Yong Qiu Shuang Cheng Jin-Yan Sun Wei Fang Simon Rayner Michael A.McVoy Xing-Jun Jiang Qiyi Tang Fang-Cheng Li Fei Hu min-hua luo Joint Center of Translational Precision Medicine Guangzhou Institute of PediatricsGuangzhou Women and Children's Medical CenterGuangzhou 510623China State Key Laboratory of Virology CAS Center for Excellence in Brain Science and Itelligence TechnologyWuhan Instute of VirologyWuhan 430071China Chinese Institute for Brain Research Beijing 102206China Wuhan Brain Hospital Ministry of TransportationWuhan 430010China Depatment of Medical Genetics Oslo University Hospital and University of OsloOsloNorway Department of Pediatrics Virginia Commonwealth University School of MedicineRichmondVirginiaUSA Department of Neurosurgery Xiangya HospitalCentral South UniversityChangha 410008China Department of Microbiology Howard University College of MedicineWashingtonDCUSA University of Chinese Academy of Sciences Bejing 100049China

Dear Editor,Gliomas are the most common brain tumors in adults which encompass all primary central nervous system(CNS)tumors of glial cell *** World Health Organization(WHO)classifies gliomas into four grades based on the histologic/prognostic *** of the unclear etiol-ogy and pathogenesis,therapeutic efficacy and prognosis is poor.

关键词： glioma prognosis pathogenesis

Restoration of FMRP expression in adult V1 neurons rescues visual deficits in a mouse model of fragile X syndrome

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Protein & Cell 2022年第3期13卷 203-219页

作者： Chaojuan Yang Yonglu Tian Feng Su Yangzhen Wang Mengna Liu Hongyi Wang Yaxuan Cui Peijiang Yuan Xiangning Li Anan Li Hui Gong Qingming luo Desheng Zhu Peng Cao Yunbo Liu Xunli Wang min-hua luo Fuqiang Xu Wei Xiong Liecheng Wang Xiang-yao Li Chen Zhang State Key Laboratory of Membrane Biology PKU-IDG/McGovern Institute for Brain ResearchSchool of Life SciencesPeking UniversityBeijing 100871China School of Basic Medical Sciences Beijing Key Laboratory of Neural Regeneration and RepairCapital Medical UniversityBeijing 100069China Peking-Tsinghua Center for Life Sciences Academy for Advanced Interdisciplinary StudiesPeking UniversityBeijing 100871China School of Mechanical Engineering and Automation Beihang UniversityBeijing 100191China School of Life Sciences Tsinghua UniversityBeijing 100084China Britton Chance Center for Biomedical Photonics Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and TechnologyWuhan 430074China State Key Laboratory of Brain and Cognitive Sciences Institute of BiophysicsChinese Academy of Sciences(CAS)Beijing 100101China Institute of Laboratory Animal Science Peking Union Medical College/Chinese Academy of Medical ScienceBeijing 100021China Laboratory Animal Center Fujian University of Tradition Chinese MedicineFuzhou 350122China State Key Laboratory of Virology CAS Center for Excellence in Brain Science andIntelligence TechnologyWuhan Institute of VirologyCASWuhan 430071China Center for Brain Science Key Laboratory of Magnetic Resonance in Biological Systems and State Key Laboratory of Magnetic Resonance and Atomic and Molecular PhysicsWuhan Institute of Physics and MathematicsCASCenter for Excellence in Brain Science and Intelligence TechnologyChinese Academy of SciencesWuhan 430071China Department of Physiology Anhui Medical UniversityHefei 230032China Department of Physiology Institute of Neuroscience and Collaborative Innovation Center for Brain ScienceSchool of MedicineZhejiang UniversityHangzhou 310058China

Many people affected by fragile X syndrome(FXS)and autism spectrum disorders have sensory processing deficits,such as hypersensitivity to auditory,tactile,and visual *** FXS in humans,loss of Fmr1 in rodents also cause sensory,behavioral,and cognitive ***,the neural mechanisms underlying sensory impairment,especially vision impairment,remain *** remains elusive whether the visual processing deficits originate from corrupted inputs,impaired perception in the primary sensory cortex,or altered integration in the higher cortex,and there is no effective *** this study,we used a genetic knockout mouse model(Fmr1^(KO)),in vivo imaging,and behavioral measurements to show that the loss of Fmr1 impaired signal processing in the primary visual cortex(V1).Specifically,Fmr1^(KO) mice showed enhanced responses to low-intensity stimuli but normal responses to high-intensity *** abnormality was accompanied by enhancements in local network connectivity in V1 microcircuits and increased dendritic complexity of V1 *** effects were ameliorated by the acute application of GABAA receptor activators,which enhanced the activity of inhibitory neurons,or by reintroducing Fmr1 gene expression in knockout V1 neurons in both juvenile and young-adult ***,V1 plays an important role in the visual abnormalities of Fmr1^(KO) mice and it could be possible to rescue the sensory disturbances in developed FXS and autism patients.

关键词： autism spectrum disorder calcium imaging fragile X syndrome primary visual cortex visual hypersensitivity