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Natacha Merindol, Mohamed El-Far, Mohamed Sylla, Nasser Masroori, Caroline Dufour, Jia-Xin Li, Pearl Cherry, Mélodie B Plourde, Cécile Tremblay, Lionel Berthoux
Elite controllers (ECs) are a rare subset of HIV-1 slow progressors characterized by prolonged viremia suppression. HLA alleles B27 and B57 promote the cytotoxic T lymphocyte (CTL)-mediated depletion of infected cells in ECs, leading to the emergence of escape mutations in the viral capsid (CA). Whether those mutations modulate CA detection by innate sensors and effectors is poorly known. Here, we investigated the targeting of CA from B27/B57+ individuals by cytosolic antiviral factors Mx2 and TRIM5α. Toward that aim, we constructed chimeric HIV-1 vectors using CA isolated from B27/B57+ or control subjects...
November 12, 2018: PLoS Pathogens
Franziska Jönsson, Claudia Hagedorn, Florian Kreppel
Adenovirus vectors are potent tools for genetic vaccination and oncolytic virotherapy. However, they are prone to multiple undesired vector-host interactions, especially after in vivo delivery. It is a consensus that the limitations imposed by undesired vector-host interactions can only be overcome if defined modifications of the vector surface are performed. These modifications include shielding of the particles from unwanted interactions and targeting by the introduction of new ligands. The goal of the protocol presented here is to enable the reader to generate shielded and, if desired, retargeted human adenovirus gene transfer vectors or oncolytic viruses...
October 26, 2018: Journal of Visualized Experiments: JoVE
Mohan Kumar Haleyur Giri Setty, Aditya Kurdekar, Prerna Mahtani, Jikun Liu, Indira Hewlett
BACKGROUND: Accurate and early detection of diverse HIV-1 subtypes using currently available assays has been a major challenge. We report development of a sensitive Time Resolved Fluorescence (TRF) Europium Nanoparticle Immuno Assay (ENIA) for cross subtype detection of p24 antigen using broadly cross reactive antibodies. METHODS: Several antibodies were tested for optimal reactivity with antigens of diverse HIV-1 subtypes and CRFs. We tested HIV strains using this assay for sensitivity and quantification ability at the pico-gram (pg)/ml level...
November 9, 2018: AIDS Research and Human Retroviruses
Maria Yanqing Chen, Susan S Butler, Weitong Chen, Junghae Suh
The fields of physical, chemical, and synthetic virology work in partnership to reprogram viruses as controllable nanodevices. Physical virology provides the fundamental biophysical understanding of how virus capsids assemble, disassemble, display metastability, and assume various configurations. Chemical virology considers the virus capsid as a chemically addressable structure, providing chemical pathways to modify the capsid exterior, interior, and subunit interfaces. Synthetic virology takes an engineering approach, modifying the virus capsid through rational, combinatorial, and bioinformatics-driven design strategies...
November 8, 2018: Wiley Interdisciplinary Reviews. Nanomedicine and Nanobiotechnology
Aakriti Alisha Arora, Chamaree de Silva
Since the development of DNA origami by Paul Rothemund in 2006, the field of structural DNA nanotechnology has undergone tremendous growth. Through DNA origami and related approaches, self-assembly of specified DNA sequences allows for the 'bottom-up' construction of diverse nanostructures. By utilizing different sets of small 'staple' DNA strands to direct the folding of a long scaffold strand in diverse ways, DNA origami has particularly been incorporated into a variety of prototypical applications beyond the two-dimensional (2D) smiley face...
2018: Nano Reviews & Experiments
Rosemary J Redfield, Shannon M Soucy
Bacterial gene transfer agents (GTAs) are small virus-like particles that package DNA fragments and inject them into cells. They are encoded by gene clusters resembling defective prophages, with genes for capsid head and tail components. These gene clusters are usually assumed to be maintained by selection for the benefits of GTA-mediated recombination, but this has never been tested. We rigorously examined the potential benefits of GTA-mediated recombination, considering separately transmission of GTA-encoding genes and recombination of all chromosomal genes...
2018: Frontiers in Microbiology
Marta Jedynak, Remigiusz Worch, Małgorzata Podsiadła-Białoskórska, Jadwiga Chroboczek, Ewa Szołajska
Adenoviral dodecahedron is a virus-like particle composed of twelve penton base proteins, derived from the capsid of human adenovirus type 3. Due to the high cell penetration capacity, it was used as a vector for protein, peptide and drug delivery. Two receptors are known to be involved in the endocytic dodecahedron uptake, namely αv integrins and heparan sulfate proteoglycans. Since it has been observed, that dodecahedron efficiently penetrates a wide range of cancer cells, it suggests that other cellular compounds may play a role in the particle endocytosis...
November 2018: Biochimica et biophysica acta. Biomembranes
Jaison Jeevanandam, Kaushik Pal, Michael K Danquah
Viruses are considered as natural nanomaterials as they are in the size range of 20-500 nm with a genetical material either DNA or RNA, which is surrounded by a protein coat capsid. Recently, the field of virus nanotechnology is gaining significant attention from researchers. Attention is given to the utilization of viruses as nanomaterials for medical, biotechnology and energy applications. Removal of genetic material from the viral capsid creates empty capsid for drug incorporation and coating the capsid protein crystals with antibodies, enzymes or aptamers will enhance their targeted drug deliver efficiency...
November 5, 2018: Biochimie
Honghui Liu, Hany Geagea, Geneviève M Rousseau, Simon J Labrie, Denise M Tremblay, Xinchun Liu, Sylvain Moineau
The virulent phage ST32 that infects the Escherichia coli strain ST130 was isolated from a wastewater sample in China and analyzed. Morphological observations showed that phage ST32 belongs to the Myoviridae family, as it has an icosahedral capsid and long contractile tail. Host range analysis showed that it exhibits a broad range of hosts including non-pathogenic and pathogenic E. coli strains. Interestingly, phage ST32 had a much larger burst size when amplified at 20 °C as compared to 30 °C or 37 °C. Its double-stranded DNA genome was sequenced and found to contain 53,092 bp with a GC content of 44...
November 7, 2018: Viruses
François Rousset, Lun Cui, Elise Siouve, Christophe Becavin, Florence Depardieu, David Bikard
High-throughput genetic screens are powerful methods to identify genes linked to a given phenotype. The catalytic null mutant of the Cas9 RNA-guided nuclease (dCas9) can be conveniently used to silence genes of interest in a method also known as CRISPRi. Here, we report a genome-wide CRISPR-dCas9 screen using a starting pool of ~ 92,000 sgRNAs which target random positions in the chromosome of E. coli. To benchmark our method, we first investigate its utility to predict gene essentiality in the genome of E...
November 7, 2018: PLoS Genetics
Emily P Thi, Ammen P Dhillon, Andrzej Ardzinski, Lale Bidirici-Ertekin, Kyle D Cobarrubias, Andrea Cuconati, Andrew S Kondratowicz, Kaylyn Kwak, Alice H L Li, Angela Miller, Chris Pasetka, Luying Pei, Janet R Phelps, Nicholas M Snead, Xiaohe Wang, Xin Ye, Michael Sofia, Amy C H Lee
Current approved nucleoside analog treatments for chronic hepatitis B virus infection are effective at controlling viral titre but are not curative and have minimal impact on the production of viral proteins such as surface antigen (HBsAg), the HBV envelope protein believed to play a role in maintaining the immune tolerant state required for viral persistence. Novel agents are needed to effect HBV cure, and reduction of HBV antigenemia may potentiate activation of effective and long-lasting host immune control...
November 7, 2018: ACS Infectious Diseases
Krystal A Fontaine, Kristoffer E Leon, Mir M Khalid, Sakshi Tomar, David Jimenez-Morales, Mariah Dunlap, Julia A Kaye, Priya S Shah, Steve Finkbeiner, Nevan J Krogan, Melanie Ott
Zika virus (ZIKV) infection of neural progenitor cells (NPCs) in utero is associated with neurological disorders, such as microcephaly, but a detailed molecular understanding of ZIKV-induced pathogenesis is lacking. Here we show that in vitro ZIKV infection of human cells, including NPCs, causes disruption of the nonsense-mediated mRNA decay (NMD) pathway. NMD is a cellular mRNA surveillance mechanism that is required for normal brain size in mice. Using affinity purification-mass spectrometry, we identified multiple cellular NMD factors that bind to the viral capsid protein, including the central NMD regulator up-frameshift protein 1 (UPF1)...
November 6, 2018: MBio
Gabriella Donà, Alessandra Andrisani, Elena Tibaldi, Anna Maria Brunati, Chiara Sabbadin, Decio Armanini, Guido Ambrosini, Eugenio Ragazzi, Luciana Bordin
Astaxanthin (Asta), red pigment of the carotenoid family, is known for its anti-oxidant, anti-cancer, anti-diabetic, and anti-inflammatory properties. In this study, we evaluated the effects of Asta on isolated human sperm in the presence of human papillomavirus (HPV) 16 capsid protein, L1. Sperm, purified by gradient separation, were treated with HPV16-L1 in both a dose and time-dependent manner in the absence or presence of 30 min-Asta pre-incubation. Effects of HPV16-L1 alone after Asta pre-incubation were evaluated by rafts (CTB) and Lyn dislocation, Tyr-phosphorylation (Tyr-P) of the head, percentages of acrosome-reacted cells (ARC) and endogenous reactive oxygen species (ROS) generation...
November 2, 2018: Marine Drugs
Carmen A Dunbar, Heather M Callaway, Colin R Parrish, Martin F Jarrold
There are many techniques for monitoring and measuring the interactions between proteins and ligands. Most of these techniques are ensemble methods that can provide association constants and in some cases stoichiometry. Here we use charge detection mass spectrometry (CDMS), a single particle technique, to probe the interactions of antigen binding fragments (Fabs) from a series of antibodies with the canine parvovirus (CPV) capsid. In addition to providing the average number of bound Fabs as a function of Fab concentration (i...
November 6, 2018: Journal of the American Chemical Society
Ziying Yan, Wei Zou, Zehua Feng, Weiran Shen, Soo-Yeun Park, Xuefeng Deng, Jianming Qiu, John F Engelhardt
The genome of recombinant adeno-associated virus 2 (rAAV2) remains a promising candidate for gene therapy for cystic fibrosis (CF) lung disease, but due to limitations in the packaging capacity and the tropism of this virus with respect to the airways, strategies have evolved for packaging an rAAV2 genome (up to 5.8kb) into the capsid of human bocavirus 1 (HBoV1) to produce a chimeric rAAV2/HBoV1 vector. Although a replication-incompetent HBoV1 genome has been established as a trans helper for capsid complementation, this system remains suboptimal with respect to virion yield...
November 6, 2018: Human Gene Therapy
Zekun Wang, Fang Cheng, John F Engelhardt, Ziying Yan, Jianming Qiu
Human bocavirus 1 (HBoV1), an autonomous parvovirus, is a helper virus supporting replication of wild-type adeno-associated virus 2 (AAV2). In this study, we compared the helper functions from HBoV1 with those from adenovirus (Ad) for the production of recombinant AAV (rAAV) vector in HEK293 cells. We demonstrated that triple plasmids transfection of (1) a cloned HBoV1 helper minigenome (pBocaHelper) that expresses HBoV1 genes NP1 , NS2 , and BocaSR, (2) pAAV transfer plasmid, and (3) pAAVRepCap supports rAAV production in HEK293 cells...
December 14, 2018: Molecular Therapy. Methods & Clinical Development
Qingbing Zheng, Rui Zhu, Longfa Xu, Maozhou He, Xiaodong Yan, Dongxiao Liu, Zhichao Yin, Yangtao Wu, Yongchao Li, Lisheng Yang, Wangheng Hou, Shuxuan Li, Zizhen Li, Zhenqin Chen, Zhihai Li, Hai Yu, Ying Gu, Jun Zhang, Timothy S Baker, Z Hong Zhou, Barney S Graham, Tong Cheng, Shaowei Li, Ningshao Xia
Enterovirus D68 (EV-D68) undergoes structural transformation between mature, cell-entry intermediate (A-particle) and empty forms throughout its life cycle. Structural information for the various forms and antibody-bound capsids will facilitate the development of effective vaccines and therapeutics against EV-D68 infection, which causes childhood respiratory and paralytic diseases worldwide. Here, we report the structures of three EV-D68 capsid states representing the virus at major phases. We further describe two original monoclonal antibodies (15C5 and 11G1) with distinct structurally defined mechanisms for virus neutralization...
November 5, 2018: Nature Microbiology
Huiyan Hu, Jingjing Zhao, Wen Yu, Junwei Zhao, Zhewei Wang, Lin Jin, Yunyun Yu, Lingfei Han, Lu Wang, Huiting Zhu, Fang Li
OBJECTIVES: Cervical cancer is the most common malignant tumors in women leading to serious morbidity and mortality worldwide, especially among developing countries. A main cause of the disease is the high-risk human papillomavirus (HR-HPV) infection. HSIL usually progress to cervical cancer, and low-grade lesions, including LSIL and ASCUS, mostly turn to normal or benign lesions, but there are still a small number of patients who will progress to HSIL. Up to now there is no efficient biomarker clinically available to predict people with high risk to progress into HSIL...
November 2, 2018: Archives of Gynecology and Obstetrics
Charlotte Floderer, Jean-Baptiste Masson, Elise Boilley, Sonia Georgeault, Peggy Merida, Mohamed El Beheiry, Maxime Dahan, Philippe Roingeard, Jean-Baptiste Sibarita, Cyril Favard, Delphine Muriaux
Monitoring virus assembly at the nanoscale in host cells remains a major challenge. Human immunodeficiency virus type 1 (HIV-1) components are addressed to the plasma membrane where they assemble to form spherical particles of 100 nm in diameter. Interestingly, HIV-1 Gag protein expression alone is sufficient to produce virus-like particles (VLPs) that resemble the immature virus. Here, we monitored VLP formation at the plasma membrane of host CD4+ T cells using a newly developed workflow allowing the analysis of long duration recordings of single-molecule Gag protein localisation and movement...
November 2, 2018: Scientific Reports
Chang-Long Zheng, Yong-Mei Fu, Zhan-Xue Xu, Yong Zou, Kai Deng
Hepatitis B virus (HBV) core protein (HBc) serves pivotal roles in the viral life cycle, particularly serving as the basic unit for capsid assembly, and is closely associated with HBV genome replication and progeny virion production. Previous studies have demonstrated that HBc has at least two functional interfaces; two HBc monomers form a homodimer via an intradimer interface, and then 90 or 120 homodimers form an icosahedral capsid via a dimer‑dimer interface. In the present study, the role of the HBc dimer‑dimer interface in HBV replication was investigated...
November 2, 2018: Molecular Medicine Reports
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