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https://www.readbyqxmd.com/read/28521201/validation-of-the-use-of-an-artificial-mitochondrial-reporter-dna-vector-containing-a-cytomegalovirus-promoter-for-mitochondrial-transgene-expression
#1
Yuma Yamada, Takuya Ishikawa, Hideyoshi Harashima
Mitochondria have their own gene expression system that is independent of the nuclear system, and control cellular functions in cooperation with the nucleus. While a number of useful technologies for achieving nuclear transgene expression have been reported, only a few have focused on mitochondria. In this study, we validated the utility of an artificial mitochondrial DNA vector with a virus promoter on mitochondrial transgene expression. We designed and constructed pCMV-mtLuc (CGG) that contains a CMV promotor derived from Cytomegalovirus and an artificial mitochondrial genome with a NanoLuc (Nluc) luciferase gene that records adjustments to the mitochondrial codon system...
May 10, 2017: Biomaterials
https://www.readbyqxmd.com/read/28510221/development-of-a-dual-expression-vector-facilitated-with-selection-free-pcr-recombination-cloning-strategy
#2
Liting Cao, Yancheng Zhou, Lin Huang, Shiqi Dong, Yue Ma
The conventional procedure for the construction of recombinant expression vector of a target gene includes PCR cloning and restriction enzyme mediated subcloning, which is time-consuming and sometimes troublesome because of the inefficiency of ligation. A variety of ligase-independent PCR cloning strategies have been developed, but they either involve complicated PCR procedures or need other DNA modifying enzymes. In this study, we report the design, and construction of an omnipotent expression vector pOmni, with which a target gene can be easily cloned through innovative selection-free PCR recombination cloning strategy with only one pair of primer and two times of PCR in one work day, without using any restriction enzymes, ligase and other DNA modifying enzymes...
December 2017: AMB Express
https://www.readbyqxmd.com/read/28508224/rescue-of-infectious-salmon-anemia-virus-isav-from-cloned-cdna
#3
Daniela Toro-Ascuy, Marcelo Cortez-San Martín
The piscine Orthomyxovirus called Infectious Salmon Anemia Virus (ISAV) is one of the most important emerging pathogens affecting the salmon industry worldwide. The first reverse genetics system for ISAV, which allows the generation of recombinant ISA virus (rISAV), is an important tool for the characterization and study of this fish virus. The plasmid-based reverse genetics system for ISAV includes the use of a novel fish promoter, the Atlantic salmon internal transcribed spacer region 1 (ITS-1). The salmon, viral and mammalian genetic elements included in pSS-URG vectors allow the expression of the eight viral RNA segments...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28508212/reverse-genetics-for-porcine-reproductive-and-respiratory-syndrome-virus
#4
Mingyuan Han, Hanzhong Ke, Yijun Du, Qingzhan Zhang, Dongwan Yoo
Porcine reproductive and respiratory syndrome (PRRS) is a reemerging swine disease, and has become economically the most significant disease in pork production worldwide. The causative agent is PRRS virus (PRRSV), which is a member virus of the family Arteriviridae. The PRRSV genome is a single-stranded positive-sense RNA and is infectious. Two strategies in the PRRSV reverse genetics system have been employed for reconstitution of progeny virus: RNA transfection and DNA transfection. The PRRSV reverse genetics has broadly been used for studies including protein structure-function relationship, foreign gene expression, vaccine development, virulence determinants, and viral pathogenesis...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28497804/vaccine-based-immunotherapeutic-approaches-to-gliomas-and-beyond
#5
REVIEW
Michael Weller, Patrick Roth, Matthias Preusser, Wolfgang Wick, David A Reardon, Michael Platten, John H Sampson
Astrocytic and oligodendroglial gliomas are intrinsic brain tumours characterized by infiltrative growth and resistance to classic cancer therapies, which renders them inevitably lethal. Glioblastoma, the most common type of glioma, also exhibits neoangiogenesis and profound immunosuppressive properties. Accordingly, strategies to revert glioma-associated immunosuppression and promote tumour-directed immune responses have been extensively explored in rodent models and in large clinical trials of tumour immunotherapy...
May 12, 2017: Nature Reviews. Neurology
https://www.readbyqxmd.com/read/28491825/hcmv-activation-of-erk-mapk-drives-a-multi-factorial-response-promoting-the-survival-of-infected-myeloid-progenitors
#6
Verity Kew, Mark Wills, Matthew Reeves
Viral binding and entry provides the first trigger of a cell death response and thus how human cytomegalovirus (HCMV) evades this - particularly during latent infection where a very limited pattern of gene expression is observed - is less well understood. It has been demonstrated that the activation of cellular signalling pathways upon virus binding promotes the survival of latently infected cells by the activation of cell encoded anti-apoptotic responses. In CD34+ cells, a major site of HCMV latency, ERK signalling is important for survival and we now show that the activation of this pathway impacts on multiple aspects of cell death pathways...
2017: Journal of Molecular Biochemistry
https://www.readbyqxmd.com/read/28480312/spatially-and-temporally-regulated-nrf2-gene-therapy-using-mcp-1-promoter-in-retinal-ganglion-cell-injury
#7
Kosuke Fujita, Koji M Nishiguchi, Yukihiro Shiga, Toru Nakazawa
Retinal ganglion cell degeneration triggered by axonal injury is believed to underlie many ocular diseases, including glaucoma and optic neuritis. In these diseases, retinal ganglion cells are affected unevenly, both spatially and temporally, such that healthy and unhealthy cells coexist in different patterns at different time points. Herein, we describe a temporally and spatially regulated adeno-associated virus gene therapy aiming to reduce undesired off-target effects on healthy retinal neurons. The Mcp-1 promoter previously shown to be activated in stressed retinal ganglion cells following murine optic nerve injury was combined with the neuroprotective intracellular transcription factor Nrf2...
June 16, 2017: Molecular Therapy. Methods & Clinical Development
https://www.readbyqxmd.com/read/28473657/human-cytomegalovirus-immediate-early-protein-promotes-survival-of-glioma-cells-through-interacting-and-acetylating-atf5
#8
Ming Hu, Bin Wang, Dongmeng Qian, Mengyuan Wang, Rui Huang, Li Wei, Ling Li, Li Zhang, David X Liu
Human cytomegalovirus (HCMV), a widespread beta-herpes virus, infects a high percentage of gliomas. HCMV is specifically detected in human gliomas at a low level of expression raises the possibility that it may regulate the malignant phenotype in a chronic manner. Although HCMV is not recognized as an oncogenic virus, it might dysregulate signaling pathways involved in initiation and promotion of malignancy.Here, our immunohistochemical staining reveals that nucleus staining of the HCMV 86-kDa immediate-early protein (IE86) is markedly increased in GBM (58...
May 9, 2017: Oncotarget
https://www.readbyqxmd.com/read/28469666/anatomical-distributional-defects-in-mutant-genes-associated-with-dominant-intermediate-charcot-marie-tooth-disease-type-c-in-an-adenovirus-mediated-mouse-model
#9
SeoJin Lee, Sandesh Panthi, Hyun Woo Jo, Jaeyoung Cho, Min-Sik Kim, Na Young Jeong, In Ok Song, Junyang Jung, Youngbuhm Huh
Dominant intermediate Charcot-Marie-Tooth disease type C (DI-CMTC) is a dominantly inherited neuropathy that has been classified primarily based on motor conduction velocity tests but is now known to involve axonal and demyelination features. DI-CMTC is linked to tyrosyl-tRNA synthetase (YARS)-associated neuropathies, which are caused by E196K and G41R missense mutations and a single de novo deletion (153-156delVKQV). It is well-established that these YARS mutations induce neuronal dysfunction, morphological symptoms involving axonal degeneration, and impaired motor performance...
March 2017: Neural Regeneration Research
https://www.readbyqxmd.com/read/28466469/recipient-donor-kir-ligand-matching-prevents-cmv-reactivation-post-haploidentical-t-cell-replete-transplantation
#10
Xiang-Yu Zhao, Xue-Yi Luo, Xing-Xing Yu, Xiao-Su Zhao, Ting-Ting Han, Ying-Jun Chang, Ming-Rui Huo, Lan-Ping Xu, Xiao-Hui Zhang, Kai-Yan Liu, Dan Li, Zheng-Fan Jiang, Xiao-Jun Huang
Licensed natural killer (NK) cells have been demonstrated to have anti-cytomegalovirus (CMV) activity. We prospectively analysed the human leucocyte antigen typing of donor-recipient pairs and the killer cell immunoglobulin-like receptor (KIR) typing of donors for 180 leukaemia patients to assess the predictive roles of licensed NK cells on CMV reactivation post-T-cell-replete haploidentical stem cell transplantation. Multivariate analysis showed that donor-recipient KIR ligand graft-versus-host or host-versus-graft direction mismatch was associated with increased refractory CMV infection (Hazard ratio = 2·556, 95% confidence interval, 1·377-4·744, P = 0·003) post-transplantation...
May 3, 2017: British Journal of Haematology
https://www.readbyqxmd.com/read/28456604/herpesviruses-hijack-host-exosomes-for-viral-pathogenesis
#11
REVIEW
Sara Sadeghipour, Rommel A Mathias
Herpesviruses are remarkable pathogens possessing elaborate mechanisms to seize various host cellular components for immune evasion, replication, and virion egress. As viruses are dependent upon their hosts, investigating this intricate interplay has revealed that the exosome pathway is utilised by alpha (Herpes Simplex Virus 1), beta (Human Cytomegalovirus, and Human Herpesvirus 6) and gamma (Epstein-Barr Virus, and Kaposi Sarcoma-associated Herpesvirus) herpesviruses. Virions and exosomes share similar properties and functions...
April 26, 2017: Seminars in Cell & Developmental Biology
https://www.readbyqxmd.com/read/28442010/-effect-of-nm23-h1-nuclear-localization-on-proliferation-of-%C3%A2-human-lung-adenocarcinoma-cell-line-a549
#12
Ya Sheng, Yanli Xiong, Mingfang Xu, Xunjie Kuang, Dong Wang, Xueqin Yang
BACKGROUND: Recent studies have indicated that Nm23-H1 is found in the nucleus, but previous studies have been based on the overexpression or suppression of Nm23-H1 in the cytoplasm. Due to the lacking nuclear localization signal of Nm23-H1, these results cannot reflect or repeat cells in which Nm23-H1 mainly positioned in nuclei and whether they cause clinical biological effects. Therefore, to explore the effects of transposing Nm23-H1 from the cytoplasm to the nucleus during lung cancer cell proliferation, a vector with a nuclear localization signal of Nm23-H1 was constructed and A549 cells were transfected...
April 20, 2017: Zhongguo Fei Ai za Zhi, Chinese Journal of Lung Cancer
https://www.readbyqxmd.com/read/28427998/nitric-oxide-regulated-proteolysis-of-human-cyp2b6-via-the-ubiquitin-proteasome-system
#13
Choon-Myung Lee, Shweta Tripathi, Edward T Morgan
We showed previously that rat cytochrome P450 CYP2B1 undergoes NO-dependent proteasomal degradation in response to inflammatory stimuli, and that the related human enzyme CYP2B6 is also down-regulated by NO in primary human hepatocytes. To investigate the mechanism of CYP2B6 down-regulation, we made several cell lines (HeLa and HuH7 cells) in which native CYP2B6 or CYP2B6 with a C-terminal V5 tag (CYP2B6V5) are expressed from a lentiviral vector with a cytomegalovirus promoter. Native CYP2B6 protein was rapidly down-regulated in HeLa cells within 3h of treatment with the NO donor (Z)-1-[2-(2-Aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate, while its mRNA level was not down-regulated...
April 17, 2017: Free Radical Biology & Medicine
https://www.readbyqxmd.com/read/28420741/new-mechanism-by-which-human-cytomegalovirus-micrornas-negate-the-proinflammatory-response-to-infection
#14
Andrew D Yurochko
Viruses have evolved many novel mechanisms to promote infection and to mitigate the host cell response to that infection. In the article by M. H. Hancock et al. (mBio 8:e00109-17, 2017, https://doi.org/10.1128/mBio.00109-17), the authors describe a new mechanism by which human cytomegalovirus (HCMV) microRNAs (miRNAs; miR-US5-1 and miR-UL112-3p) negate the proinflammatory response to infection. The authors document that these two viral miRNAs downregulate the NF-κB response through direct targeting of the IKKα and IKKβ mRNAs, which in turn, through diminished IκB kinases (IKKs), block production of proinflammatory cytokines (interleukin-6 [IL-6], CCL5, and tumor necrosis factor alpha [TNF-α])...
April 18, 2017: MBio
https://www.readbyqxmd.com/read/28420570/herpesviradae-infections-in-severely-burned-children
#15
Paul Wurzer, Megan R Cole, Robert P Clayton, Gabriel Hundeshagen, Omar Nunez Lopez, Janos Cambiaso-Daniel, Raimund Winter, Ludwik K Branski, Hal K Hawkins, Celeste C Finnerty, David N Herndon, Jong O Lee
OBJECTIVE: Burn-related immunosuppression can promote human herpesviridae infections. However, the effect of these infections on morbidity and mortality after pediatric burn injuries is unclear. METHODS: We retrospectively analyzed pediatric patients with burns ≥10% of the total body surface area (TBSA) who were admitted between 2010 and 2015. On clinical suspicion of a viral infection, antiviral therapy was initiated. Viral infection was confirmed via Tzanck smear, viral culture, and/or PCR...
April 15, 2017: Burns: Journal of the International Society for Burn Injuries
https://www.readbyqxmd.com/read/28411277/long-term-survival-in-glioblastoma-with-cytomegalovirus-pp65-targeted-vaccination
#16
Kristen A Batich, Elizabeth A Reap, Gary E Archer, Luis Sanchez-Perez, Smita K Nair, Robert J Schmittling, Pam Norberg, Weihua Xie, James E Herndon, Patrick Healy, Roger E McLendon, Allan H Friedman, Henry S Friedman, Darell Bigner, Gordana Vlahovic, Duane A Mitchell, John H Sampson
Purpose: Patients with glioblastoma have less than 15-month median survival despite surgical resection, high-dose radiation, and chemotherapy with temozolomide. We previously demonstrated that targeting cytomegalovirus pp65 using dendritic cells (DC) can extend survival and, in a separate study, that dose-intensified temozolomide (DI-TMZ) and adjuvant granulocyte macrophage colony-stimulating factor (GM-CSF) potentiate tumor-specific immune responses in patients with glioblastoma. Here, we evaluated pp65-specific cellular responses following DI-TMZ with pp65-DCs and determined the effects on long-term progression-free survival (PFS) and overall survival (OS)...
April 15, 2017: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/28409504/comparative-study-of-the-transfection-efficiency-of-commonly-used-viral-vectors-in-rhesus-monkey-macaca-mulatta-brains
#17
Shi-Hao Wu, Zhi-Xing Liao, Joshua D Rizak, Na Zheng, Lin-Heng Zhang, Hen Tang, Xiao-Bin He, Yang Wu, Xia-Ping He, Mei-Feng Yang, Zheng-Hui Li, Dong-Dong Qin, Xin-Tian Hu
Viral vector transfection systems are among the simplest of biological agents with the ability to transfer genes into the central nervous system. In brain research, a series of powerful and novel gene editing technologies are based on these systems. Although many viral vectors are used in rodents, their full application has been limited in non-human primates. To identify viral vectors that can stably and effectively express exogenous genes within non-human primates, eleven commonly used recombinant adeno-associated viral and lentiviral vectors, each carrying a gene to express green or red fluorescence, were injected into the parietal cortex of four rhesus monkeys...
March 18, 2017: Zoological Research
https://www.readbyqxmd.com/read/28407741/the-pd-1-pd-l1-pathway-promotes-development-of-brain-resident-memory-t-cells-following-acute-viral-encephalitis
#18
Sujata Prasad, Shuxian Hu, Wen S Sheng, Priyanka Chauhan, Amar Singh, James R Lokensgard
BACKGROUND: Previous work from our laboratory has demonstrated that during acute viral brain infection, glial cells modulate antiviral T cell effector responses through the PD-1: PD-L1 pathway, thereby limiting the deleterious consequences of unrestrained neuroinflammation. Here, we evaluated the PD-1: PD-L1 pathway in development of brain-resident memory T cells (bTRM) following murine cytomegalovirus (MCMV) infection. METHODS: Flow cytometric analysis of immune cells was performed at 7, 14, and 30 days post-infection (dpi) to assess the shift of brain-infiltrating CD8(+) T cell populations from short-lived effector cells (SLEC) to memory precursor effector cells (MPEC), as well as generation of bTRMs...
April 13, 2017: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/28404844/intramuscular-immunization-of-mice-with-the-live-attenuated-herpes-simplex-virus-type-1-hsv-1-vaccine-strain-vc2-expressing-the-equine-herpesvirus-1-ehv-1-glycoprotein-d-gd-generates-anti-ehv-1-immune-responses-in-mice
#19
Shiliang A Liu, Brent A Stanfield, Vladimir N Chouljenko, Shan Naidu, Ingeborg Langohr, Fabio Del Piero, Jacqueline Ferracone, Alma A Roy, Konstantin G Kousoulas
Vaccination remains the best option to combat EHV-1 infection and several different strategies of vaccination have been investigated and developed over the past decades. Herein, we report that the live-attenuated Herpes Simplex Virus Type-1 (HSV-1) VC2 vaccine strain, which has been shown to be unable to enter into neurons and establish latency in mice, can be utilized as a vector for the heterologous expression of the Equine Herpesvirus-1 (EHV-1) glycoprotein D (gD), and that intramuscular immunization of mice resulted in strong anti-viral humoral and cellular immune responses...
April 12, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28403202/human-cytomegalovirus-glycoprotein-complex-gh-gl-go-uses-pdgfr-%C3%AE-as-a-key-for-entry
#20
Yiquan Wu, Adrian Prager, Simone Boos, Moritz Resch, Ilija Brizic, Michael Mach, Sabrina Wildner, Laura Scrivano, Barbara Adler
Herpesvirus gH/gL envelope glycoprotein complexes are key players in virus entry as ligands for host cell receptors and by promoting fusion of viral envelopes with cellular membranes. Human cytomegalovirus (HCMV) has two alternative gH/gL complexes, gH/gL/gO and gH/gL/UL128,130,131A which both shape the HCMV tropism. By studying binding of HCMV particles to fibroblasts, we could for the first time show that virion gH/gL/gO binds to platelet-derived growth factor-α (PDGFR-α) on the surface of fibroblasts and that gH/gL/gO either directly or indirectly recruits gB to this complex...
April 2017: PLoS Pathogens
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