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1.
Front Immunol ; 12: 727046, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34691033

RESUMO

Vaccinia virus (VV) is the most studied member of the poxvirus family, is responsible for the successful elimination of smallpox worldwide, and has been developed as a vaccine vehicle for infectious diseases and cancer immunotherapy. We have previously shown that the unique potency of VV in the activation of CD8+ T cell response is dependent on efficient activation of the innate immune system through Toll-like receptor (TLR)-dependent and -independent pathways. However, it remains incompletely defined what regulate CD8+ T cell response to VV infection. In this study, we showed that γδT cells play an important role in promoting CD8+ T cell response to VV infection. We found that γδT cells can directly present viral antigens in the context of MHC-I for CD8+ T cell activation to VV in vivo, and we further demonstrated that cell-intrinsic MyD88 signaling in γδT cells is required for activation of γδT cells and CD8+ T cells. These results illustrate a critical role for γδT cells in the regulation of adaptive T cell response to viral infection and may shed light on the design of more effective vaccine strategies based on manipulation of γδT cells.


Assuntos
Linfócitos T CD8-Positivos/imunologia , Linfócitos Intraepiteliais/imunologia , Vaccinia/imunologia , Animais , Apresentação do Antígeno , Antígenos Virais/imunologia , Camundongos Endogâmicos C57BL , Camundongos Knockout , Vírus Vaccinia
2.
Emerg Infect Dis ; 27(7): 1989-1991, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34152972

RESUMO

Buffalopox outbreaks caused by vaccinia virus were observed in villages of Tamil Nadu, India, among lactating buffaloes and cows. Milkers also had lesions on their fingers. Because vaccinia virus is known to have extended its host range in Brazil, we recommend continuous surveillance to understand cross-species transmission and to curtail disease effects.


Assuntos
Gado , Vaccinia , Animais , Brasil , Bovinos , Surtos de Doenças , Feminino , Índia , Lactação , Vaccinia/epidemiologia , Vírus Vaccinia , Zoonoses/epidemiologia
3.
Front Immunol ; 12: 645210, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33959127

RESUMO

Vaccination is one of the most efficient public healthcare measures to fight infectious diseases. Nevertheless, the immune mechanisms induced in vivo by vaccination are still unclear. The route of administration, an important vaccination parameter, can substantially modify the quality of the response. How the route of administration affects the generation and profile of immune responses is of major interest. Here, we aimed to extensively characterize the profiles of the innate and adaptive response to vaccination induced after intradermal, subcutaneous, or intramuscular administration with a modified vaccinia virus Ankara model vaccine in non-human primates. The adaptive response following subcutaneous immunization was clearly different from that following intradermal or intramuscular immunization. The subcutaneous route induced a higher level of neutralizing antibodies than the intradermal and intramuscular vaccination routes. In contrast, polyfunctional CD8+ T-cell responses were preferentially induced after intradermal or intramuscular injection. We observed the same dichotomy when analyzing the early molecular and cellular immune events, highlighting the recruitment of cell populations, such as CD8+ T lymphocytes and myeloid-derived suppressive cells, and the activation of key immunomodulatory gene pathways. These results demonstrate that the quality of the vaccine response induced by an attenuated vaccine is shaped by early and subtle modifications of the innate immune response. In this immunization context, the route of administration must be tailored to the desired type of protective immune response. This will be achieved through systems vaccinology and mathematical modeling, which will be critical for predicting the efficacy of the vaccination route for personalized medicine.


Assuntos
Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Linfócitos T CD8-Positivos/imunologia , Células Supressoras Mieloides/imunologia , Vacinação , Vírus Vaccinia/imunologia , Vaccinia/imunologia , Vacinas Virais/farmacologia , Animais , Injeções Intradérmicas , Injeções Intramusculares , Macaca fascicularis , Masculino , Vacinas Atenuadas/farmacologia
4.
Immunity ; 54(5): 962-975.e8, 2021 05 11.
Artigo em Inglês | MEDLINE | ID: mdl-33857420

RESUMO

Activation of the cyclic guanosine monophosphate (GMP)-AMP (cGAMP) sensor STING requires its translocation from the endoplasmic reticulum to the Golgi apparatus and subsequent polymerization. Using a genome-wide CRISPR-Cas9 screen to define factors critical for STING activation in cells, we identified proteins critical for biosynthesis of sulfated glycosaminoglycans (sGAGs) in the Golgi apparatus. Binding of sGAGs promoted STING polymerization through luminal, positively charged, polar residues. These residues are evolutionarily conserved, and selective mutation of specific residues inhibited STING activation. Purified or chemically synthesized sGAGs induced STING polymerization and activation of the kinase TBK1. The chain length and O-linked sulfation of sGAGs directly affected the level of STING polymerization and, therefore, its activation. Reducing the expression of Slc35b2 to inhibit GAG sulfation in mice impaired responses to vaccinia virus infection. Thus, sGAGs in the Golgi apparatus are necessary and sufficient to drive STING polymerization, providing a mechanistic understanding of the requirement for endoplasmic reticulum (ER)-to-Golgi apparatus translocation for STING activation.


Assuntos
Glicosaminoglicanos/metabolismo , Complexo de Golgi/metabolismo , Proteínas de Membrana/metabolismo , Nucleotídeos Cíclicos/metabolismo , Animais , Células COS , Linhagem Celular , Linhagem Celular Tumoral , Chlorocebus aethiops , Cricetinae , Citosol/metabolismo , Retículo Endoplasmático/metabolismo , Células HeLa , Humanos , Camundongos , Polimerização , Transdução de Sinais/fisiologia , Transportadores de Sulfato/metabolismo , Vaccinia/metabolismo , Vírus Vaccinia/patogenicidade
5.
J Cell Sci ; 134(8)2021 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-33912921

RESUMO

Viral infection both activates stress signaling pathways and redistributes ribosomes away from host mRNAs to translate viral mRNAs. The intricacies of this ribosome shuffle from host to viral mRNAs are poorly understood. Here, we uncover a role for the ribosome-associated quality control (RQC) factor ZNF598 during vaccinia virus mRNA translation. ZNF598 acts on collided ribosomes to ubiquitylate 40S subunit proteins uS10 (RPS20) and eS10 (RPS10), initiating RQC-dependent nascent chain degradation and ribosome recycling. We show that vaccinia infection enhances uS10 ubiquitylation, indicating an increased burden on RQC pathways during viral propagation. Consistent with an increased RQC demand, we demonstrate that vaccinia virus replication is impaired in cells that either lack ZNF598 or express a ubiquitylation-deficient version of uS10. Using SILAC-based proteomics and concurrent RNA-seq analysis, we determine that translation, but not transcription of vaccinia virus mRNAs is compromised in cells with deficient RQC activity. Additionally, vaccinia virus infection reduces cellular RQC activity, suggesting that co-option of ZNF598 by vaccinia virus plays a critical role in translational reprogramming that is needed for optimal viral propagation.


Assuntos
Vírus Vaccinia , Vaccinia , Proteínas de Transporte/metabolismo , Células HEK293 , Humanos , Biossíntese de Proteínas , Controle de Qualidade , Ribossomos/metabolismo , Vaccinia/genética , Vírus Vaccinia/genética
6.
Virology ; 558: 22-27, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33711560

RESUMO

Cytomegalovirus (CMV) promoter drives various gene expression and yields sufficient protein for further functional investigation. Receptor binding domain (RBD) on spike protein of the SARS_CoV2 is the most critical portal for virus infection. Thus native conformational RBD protein may facilitate biochemical identification of RBD and provide valuable support of drug and vaccine design for curing COVID-19. We attempted to express RBD under CMV promoter in vitro, but failed. RBD-specific mRNA cannot be detected in cell transfected with recombinant plasmids, in which CMV promoter governs the RBD transcription. Additionally, the pCMV-Tag2B-SARS_CoV2_RBD trans-inactivates CMV promoter transcription activity. Alternatively, we identified that both Chicken ß-actin promoter and Vaccinia virus-specific medium/late (M/L) promoter (pSYN) can highly precede SARS_CoV2 RBD expression. Our findings provided evidence that SARS_CoV2 RBD gene can be driven by Chicken ß-actin promoter or Vaccinia virus-specific medium/late promoter instead of CMV promoter, thus providing valuable information for RBD feature exploration.


Assuntos
COVID-19/virologia , Citomegalovirus/genética , Regiões Promotoras Genéticas , SARS-CoV-2/genética , Glicoproteína da Espícula de Coronavírus/genética , Actinas/genética , Animais , Células CHO , Galinhas , Chlorocebus aethiops , Clonagem Molecular , Cricetulus , Células HEK293 , Humanos , Domínios Proteicos , Transcrição Genética , Vaccinia/genética , Células Vero
7.
PLoS Pathog ; 17(2): e1009303, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33529218

RESUMO

Metabolism is a crucial frontier of host-virus interaction as viruses rely on their host cells to provide nutrients and energy for propagation. Vaccinia virus (VACV) is the prototype poxvirus. It makes intensive demands for energy and macromolecules in order to build hundreds and thousands of viral particles in a single cell within hours of infection. Our comprehensive metabolic profiling reveals profound reprogramming of cellular metabolism by VACV infection, including increased levels of the intermediates of the tri-carboxylic acid (TCA) cycle independent of glutaminolysis. By investigating the level of citrate, the first metabolite of the TCA cycle, we demonstrate that the elevation of citrate depends on VACV-encoded viral growth factor (VGF), a viral homolog of cellular epidermal growth factor. Further, the upregulation of citrate is dependent on STAT3 signaling, which is activated non-canonically at the serine727 upon VACV infection. The STAT3 activation is dependent on VGF, and VGF-dependent EGFR and MAPK signaling. Together, our study reveals a novel mechanism by which VACV manipulates cellular metabolism through a specific viral factor and by selectively activating a series of cellular signaling pathways.


Assuntos
Citratos/metabolismo , Ciclo do Ácido Cítrico , Fibroblastos/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Fator de Transcrição STAT3/metabolismo , Vírus Vaccinia/fisiologia , Vaccinia/metabolismo , Receptores ErbB/genética , Receptores ErbB/metabolismo , Fibroblastos/virologia , Interações Hospedeiro-Patógeno , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Sistema de Sinalização das MAP Quinases , Metaboloma , Fosforilação , Fator de Transcrição STAT3/genética , Transdução de Sinais , Vaccinia/virologia
8.
Mem Inst Oswaldo Cruz ; 115: e200521, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33566940

RESUMO

Outbreaks of a vesiculopustular disease in dairy cattle and milkers have been frequently reported in Brazil since 1999 when the vaccinia virus strain Cantagalo was first isolated in the State of Rio de Janeiro. However, the genomic diversity of the viral isolates associated with these outbreaks is not well known, particularly in the southeastern states that represent the focal point of virus spread to other regions. Here, we report the genomic sequences and an analysis of the polymorphic site profiles and genotypic diversity of four clinical isolates of vaccinia virus strain Cantagalo collected from 1999 to 2006 in southeastern Brazil.


Assuntos
Doenças dos Bovinos , Vírus Vaccinia , Vaccinia , Animais , Brasil/epidemiologia , Bovinos , Doenças dos Bovinos/epidemiologia , Surtos de Doenças , Genômica , Filogenia , Vaccinia/epidemiologia , Vaccinia/veterinária , Vírus Vaccinia/genética
9.
J Immunol ; 206(4): 776-784, 2021 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-33419767

RESUMO

There is a paucity of information on dendritic cell (DC) responses to vaccinia virus (VACV), including the traffic of DCs to the draining lymph node (dLN). In this study, using a mouse model of infection, we studied skin DC migration in response to VACV and compared it with the tuberculosis vaccine Mycobacterium bovis bacille Calmette-Guérin (BCG), another live attenuated vaccine administered via the skin. In stark contrast to BCG, skin DCs did not relocate to the dLN in response to VACV. Infection with UV-inactivated VACV or modified VACV Ankara promoted DC movement to the dLN, indicating that interference with skin DC migration requires replication-competent VACV. This suppressive effect of VACV was capable of mitigating responses to a secondary challenge with BCG in the skin, ablating DC migration, reducing BCG transport, and delaying CD4+ T cell priming in the dLN. Expression of inflammatory mediators associated with BCG-triggered DC migration were absent from virus-injected skin, suggesting that other pathways invoke DC movement in response to replication-deficient VACV. Despite adamant suppression of DC migration, VACV was still detected early in the dLN and primed Ag-specific CD4+ T cells. In summary, VACV blocks skin DC mobilization from the site of infection while retaining the ability to access the dLN to prime CD4+ T cells.


Assuntos
Movimento Celular/imunologia , Células Dendríticas/imunologia , Linfonodos/imunologia , Pele/imunologia , Vírus Vaccinia/imunologia , Vaccinia/imunologia , Animais , Linfócitos T CD4-Positivos/imunologia , Movimento Celular/genética , Camundongos , Camundongos Knockout , Mycobacterium bovis/imunologia , Vaccinia/genética , Vírus Vaccinia/genética
10.
PLoS Pathog ; 17(1): e1009215, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33439897

RESUMO

Poxvirus systems have been extensively used as vaccine vectors. Herein a RNA-Seq analysis of intramuscular injection sites provided detailed insights into host innate immune responses, as well as expression of vector and recombinant immunogen genes, after vaccination with a new multiplication defective, vaccinia-based vector, Sementis Copenhagen Vector. Chikungunya and Zika virus immunogen mRNA and protein expression was associated with necrosing skeletal muscle cells surrounded by mixed cellular infiltrates. The multiple adjuvant signatures at 12 hours post-vaccination were dominated by TLR3, 4 and 9, STING, MAVS, PKR and the inflammasome. Th1 cytokine signatures were dominated by IFNγ, TNF and IL1ß, and chemokine signatures by CCL5 and CXCL12. Multiple signatures associated with dendritic cell stimulation were evident. By day seven, vaccine transcripts were absent, and cell death, neutrophil, macrophage and inflammation annotations had abated. No compelling arthritis signatures were identified. Such injection site vaccinology approaches should inform refinements in poxvirus-based vector design.


Assuntos
Vetores Genéticos/administração & dosagem , Imunidade Inata/imunologia , Reação no Local da Injeção/imunologia , Vacinação/métodos , Vacinas Sintéticas/administração & dosagem , Vaccinia/imunologia , Infecção por Zika virus/imunologia , Animais , Feminino , Vetores Genéticos/genética , Genoma Viral , Camundongos , Camundongos Endogâmicos C57BL , RNA-Seq , Vacinas Sintéticas/imunologia , Vaccinia/genética , Vaccinia/metabolismo , Vaccinia/virologia , Vírus Vaccinia/isolamento & purificação , Vacinologia , Zika virus/isolamento & purificação , Infecção por Zika virus/genética , Infecção por Zika virus/metabolismo , Infecção por Zika virus/virologia
11.
Immunity ; 54(2): 276-290.e5, 2021 02 09.
Artigo em Inglês | MEDLINE | ID: mdl-33434494

RESUMO

The oropharyngeal mucosa serves as a perpetual pathogen entry point and a critical site for viral replication and spread. Here, we demonstrate that type 1 innate lymphoid cells (ILC1s) were the major immune force providing early protection during acute oral mucosal viral infection. Using intravital microscopy, we show that ILC1s populated and patrolled the uninfected labial mucosa. ILC1s produced interferon-γ (IFN-γ) in the absence of infection, leading to the upregulation of key antiviral genes, which were downregulated in uninfected animals upon genetic ablation of ILC1s or antibody-based neutralization of IFN-γ. Thus, tonic IFN-γ production generates increased oral mucosal viral resistance even before infection. Our results demonstrate barrier-tissue protection through tissue surveillance in the absence of rearranged-antigen receptors and the induction of an antiviral state during homeostasis. This aspect of ILC1 biology raises the possibility that these cells do not share true functional redundancy with other tissue-resident lymphocytes.


Assuntos
Interferon gama/metabolismo , Linfócitos/imunologia , Orofaringe/imunologia , Mucosa Respiratória/imunologia , Vírus Vaccinia/fisiologia , Vaccinia/imunologia , Animais , Células Cultivadas , Resistência à Doença , Humanos , Imunidade Inata , Interferon gama/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteínas com Domínio T/genética , Células Th1/imunologia
12.
Vaccine ; 39(22): 3067-3080, 2021 05 21.
Artigo em Inglês | MEDLINE | ID: mdl-33077299

RESUMO

The Brighton Collaboration Viral Vector Vaccines Safety Working Group (V3SWG) was formed to evaluate the safety and characteristics of live, recombinant viral vector vaccines. The Modified Vaccinia Ankara (MVA) vector system is being explored as a platform for development of multiple vaccines. This paper reviews the molecular and biological features specifically of the MVA-BN vector system, followed by a template with details on the safety and characteristics of an MVA-BN based vaccine against Zaire ebolavirus and other filovirus strains. The MVA-BN-Filo vaccine is based on a live, highly attenuated poxviral vector incapable of replicating in human cells and encodes glycoproteins of Ebola virus Zaire, Sudan virus and Marburg virus and the nucleoprotein of the Thai Forest virus. This vaccine has been approved in the European Union in July 2020 as part of a heterologous Ebola vaccination regimen. The MVA-BN vector is attenuated following over 500 serial passages in eggs, showing restricted host tropism and incompetence to replicate in human cells. MVA has six major deletions and other mutations of genes outside these deletions, which all contribute to the replication deficiency in human and other mammalian cells. Attenuation of MVA-BN was demonstrated by safe administration in immunocompromised mice and non-human primates. In multiple clinical trials with the MVA-BN backbone, more than 7800 participants have been vaccinated, demonstrating a safety profile consistent with other licensed, modern vaccines. MVA-BN has been approved as smallpox vaccine in Europe and Canada in 2013, and as smallpox and monkeypox vaccine in the US in 2019. No signal for inflammatory cardiac disorders was identified throughout the MVA-BN development program. This is in sharp contrast to the older, replicating vaccinia smallpox vaccines, which have a known risk for myocarditis and/or pericarditis in up to 1 in 200 vaccinees. MVA-BN-Filo as part of a heterologous Ebola vaccination regimen (Ad26.ZEBOV/MVA-BN-Filo) has undergone clinical testing including Phase III in West Africa and is currently in use in large scale vaccination studies in Central African countries. This paper provides a comprehensive picture of the MVA-BN vector, which has reached regulatory approvals, both as MVA-BN backbone for smallpox/monkeypox, as well as for the MVA-BN-Filo construct as part of an Ebola vaccination regimen, and therefore aims to provide solutions to prevent disease from high-consequence human pathogens.


Assuntos
Vacinas contra Ebola , Vaccinia , África Ocidental , Animais , Canadá , Europa (Continente) , Camundongos , Vírus Vaccinia/genética
13.
J Virol ; 95(3)2021 01 13.
Artigo em Inglês | MEDLINE | ID: mdl-33177193

RESUMO

The poxviral B1 and B12 proteins are a homologous kinase-pseudokinase pair, which modulates a shared host pathway governing viral DNA replication and antiviral defense. While the molecular mechanisms involved are incompletely understood, B1 and B12 seem to intersect with signaling processes mediated by their cellular homologs termed the vaccinia-related kinases (VRKs). In this study, we expand upon our previous characterization of the B1-B12 signaling axis to gain insights into B12 function. We begin our studies by demonstrating that modulation of B12 repressive activity is a conserved function of B1 orthologs from divergent poxviruses. Next, we characterize the protein interactome of B12 using multiple cell lines and expression systems and discover that the cellular kinase VRK1 is a highly enriched B12 interactor. Using complementary VRK1 knockdown and overexpression assays, we first demonstrate that VRK1 is required for the rescue of a B1-deleted virus upon mutation of B12. Second, we find that VRK1 overexpression is sufficient to overcome repressive B12 activity during B1-deleted virus replication. Interestingly, we also evince that B12 interferes with the ability of VRK1 to phosphoinactivate the host defense protein BAF. Thus, B12 restricts vaccinia virus DNA accumulation in part by repressing the ability of VRK1 to inactivate BAF. Finally, these data establish that a B12-VRK1-BAF signaling axis forms during vaccinia virus infection and is modulated via kinases B1 and/or VRK2. These studies provide novel insights into the complex mechanisms that poxviruses use to hijack homologous cellular signaling pathways during infection.IMPORTANCE Viruses from diverse families encode both positive and negative regulators of viral replication. While their functions can sometimes be enigmatic, investigation of virus-encoded, negative regulators of viral replication has revealed fascinating aspects of virology. Studies of poxvirus-encoded genes have largely concentrated on positive regulators of their replication; however, examples of fitness gains attributed to poxvirus gene loss suggests that negative regulators of poxvirus replication also impact infection dynamics. This study focuses on the vaccinia B12 pseudokinase, a protein capable of inhibiting vaccinia DNA replication. Here, we elucidate the mechanisms by which B12 inhibits vaccinia DNA replication, demonstrating that B12 activates the antiviral protein BAF by inhibiting the activity of VRK1, a cellular modulator of BAF. Combined with previous data, these studies provide evidence that poxviruses govern their replication by employing both positive and negative regulators of viral replication.


Assuntos
Proteínas de Ligação a DNA/metabolismo , Interações Hospedeiro-Patógeno , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Vírus Vaccinia/enzimologia , Vaccinia/imunologia , Proteínas Virais/metabolismo , Antivirais , Proteínas de Ligação a DNA/genética , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Mutação , Fosforilação , Proteínas Serina-Treonina Quinases/genética , Transdução de Sinais , Vaccinia/metabolismo , Vaccinia/virologia , Proteínas Virais/genética
14.
Adv Mater ; 33(1): e2005448, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33230875

RESUMO

The spread of the severe acute respiratory syndrome coronavirus has changed the lives of people around the world with a huge impact on economies and societies. The development of wearable sensors that can continuously monitor the environment for viruses may become an important research area. Here, the state of the art of research on biosensor materials for virus detection is reviewed. A general description of the principles for virus detection is included, along with a critique of the experimental work dedicated to various virus sensors, and a summary of their detection limitations. The piezoelectric sensors used for the detection of human papilloma, vaccinia, dengue, Ebola, influenza A, human immunodeficiency, and hepatitis B viruses are examined in the first section; then the second part deals with magnetostrictive sensors for the detection of bacterial spores, proteins, and classical swine fever. In addition, progress related to early detection of COVID-19 (coronavirus disease 2019) is discussed in the final section, where remaining challenges in the field are also identified. It is believed that this review will guide material researchers in their future work of developing smart biosensors, which can further improve detection sensitivity in monitoring currently known and future virus threats.


Assuntos
Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Teste para COVID-19/métodos , COVID-19/diagnóstico , Magnetismo , Animais , Inteligência Artificial , Condutividade Elétrica , Infecções por HIV/diagnóstico , Doença pelo Vírus Ebola/diagnóstico , Hepatite B/diagnóstico , Humanos , Influenza Humana/diagnóstico , Infecções por Papillomavirus/diagnóstico , Dengue Grave/diagnóstico , Vaccinia/diagnóstico
15.
Rev Salud Publica (Bogota) ; 20(6): 778-783, 2020 Nov 16.
Artigo em Espanhol | MEDLINE | ID: mdl-33206905

RESUMO

The recent occurrence of vaccinia virus infections in humans and animals in Colombia, together with that reported for this and other species of the genus Orthopoxvirus in some South American, African, Asian and European countries, is supporting evidence of the emergence and re-emergence of the genus. This fact has become of great interest for public health around the world due to its biological and an epidemiological features, as was in the past the variola virus, one of its representatives. The emergence and re-emergence of the genus Orthopoxvirus may be a consequence of stopping vaccination against the variola virus in the 1970s and 1980s. This vaccination unsuspectedly induced cross-protective immunity to other species of that genus. This is a review of the history, biology and epidemiology of the main species of the genus Orthopoxvirus, together with its clinical presentation, social context and public health impact in the past, present and future.


Assuntos
Infecções por Poxviridae/epidemiologia , Zoonoses Virais/epidemiologia , Animais , Colômbia/epidemiologia , Doenças Transmissíveis Emergentes/epidemiologia , Reações Cruzadas , Erradicação de Doenças/história , Surtos de Doenças/história , Saúde Global , História do Século XVIII , História do Século XIX , História do Século XX , História do Século XXI , História Antiga , Humanos , Infecções por Poxviridae/história , Infecções por Poxviridae/imunologia , Saúde Pública , Varíola/história , Varíola/prevenção & controle , Vacina Antivariólica , Determinantes Sociais da Saúde , Vacinação , Vaccinia/epidemiologia , Zoonoses Virais/história
16.
J Cell Sci ; 133(21)2020 11 09.
Artigo em Inglês | MEDLINE | ID: mdl-33067254

RESUMO

Vaccinia-related kinase 3 (VRK3) has been reported to be a negative regulator of ERK (ERK1 and ERK2; also known as MAPK3 and MAPK1, respectively) that protects cells from persistent ERK activation and inhibits ERK-dependent apoptosis. Here we report that the E3 ubiquitin-protein ligase RNF144a promotes the degradation of VRK3 via polyubiquitylation and thus affects VRK3-mediated ERK activity. Under oxidative stress, VRK3 migrates from the nucleus to the cytoplasm, which increases its chance of interacting with RNF144a, thereby promoting the degradation of VRK3. Overexpression of RNF144a increases ERK activity via downregulation of VRK3 and promotes ERK-dependent apoptosis. In contrast, depletion of RNF144a increases the protein level of VRK3 and protects cells from excessive ERK activity. These findings suggest that VRK3 protects cells by suppressing oxidative stress-induced ERK, and that RNF144a sensitively regulates this process.


Assuntos
Vaccinia , Apoptose/genética , Proteínas de Transporte/metabolismo , Regulação para Baixo/genética , Humanos , Estresse Oxidativo/genética , Fosforilação , Proteínas Serina-Treonina Quinases , Ubiquitina-Proteína Ligases/genética
17.
PLoS Pathog ; 16(10): e1008660, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-33075093

RESUMO

Mammary carcinoma, including triple-negative breast carcinomas (TNBC) are tumor-types for which human and canine pathologies are closely related at the molecular level. The efficacy of an oncolytic vaccinia virus (VV) was compared in low-passage primary carcinoma cells from TNBC versus non-TNBC. Non-TNBC cells were 28 fold more sensitive to VV than TNBC cells in which VV replication is impaired. Single-cell RNA-seq performed on two different TNBC cell samples, infected or not with VV, highlighted three distinct populations: naïve cells, bystander cells, defined as cells exposed to the virus but not infected and infected cells. The transcriptomes of these three populations showed striking variations in the modulation of pathways regulated by cytokines and growth factors. We hypothesized that the pool of genes expressed in the bystander populations was enriched in antiviral genes. Bioinformatic analysis suggested that the reduced activity of the virus was associated with a higher mesenchymal status of the cells. In addition, we demonstrated experimentally that high expression of one gene, DDIT4, is detrimental to VV production. Considering that DDIT4 is associated with a poor prognosis in various cancers including TNBC, our data highlight DDIT4 as a candidate resistance marker for oncolytic poxvirus therapy. This information could be used to design new generations of oncolytic poxviruses. Beyond the field of gene therapy, this study demonstrates that single-cell transcriptomics can be used to identify cellular factors influencing viral replication.


Assuntos
Neoplasias Mamárias Animais/metabolismo , Terapia Viral Oncolítica/métodos , Fatores de Transcrição/metabolismo , Transcriptoma , Vírus Vaccinia/genética , Vaccinia/metabolismo , Replicação Viral , Animais , Biologia Computacional , Cães , Feminino , Neoplasias Mamárias Animais/genética , Neoplasias Mamárias Animais/terapia , Neoplasias Mamárias Animais/virologia , Análise de Célula Única , Fatores de Transcrição/genética , Vaccinia/genética , Vaccinia/virologia
18.
J Virol ; 94(24)2020 11 23.
Artigo em Inglês | MEDLINE | ID: mdl-32999026

RESUMO

We conducted an exhaustive search for three-dimensional structural homologs to the proteins of 20 key phylogenetically distinct nucleocytoplasmic DNA viruses (NCLDV). Structural matches covered 429 known protein domain superfamilies, with the most highly represented being ankyrin repeat, P-loop NTPase, F-box, protein kinase, and membrane occupation and recognition nexus (MORN) repeat. Domain superfamily diversity correlated with genome size, but a diversity of around 200 superfamilies appeared to correlate with an abrupt switch to paralogization. Extensive structural homology was found across the range of eukaryotic RNA polymerase II subunits and their associated basal transcription factors, with the coordinated gain and loss of clusters of subunits on a virus-by-virus basis. The total number of predicted endonucleases across the 20 NCLDV was nearly quadrupled from 36 to 132, covering much of the structural and functional diversity of endonucleases throughout the biosphere in DNA restriction, repair, and homing. Unexpected findings included capsid protein-transcription factor chimeras; endonuclease chimeras; enzymes for detoxification; antimicrobial peptides and toxin-antitoxin systems associated with symbiosis, immunity, and addiction; and novel proteins for membrane abscission and protein turnover.IMPORTANCE We extended the known annotation space for the NCLDV by 46%, revealing high-probability structural matches for fully 45% of the 9,671 query proteins and confirming up to 98% of existing annotations per virus. The most prevalent protein families included ankyrin repeat- and MORN repeat-containing proteins, many of which included an F-box, suggesting extensive host cell modulation among the NCLDV. Regression suggested a minimum requirement for around 36 protein structural superfamilies for a viable NCLDV, and beyond around 200 superfamilies, genome expansion by the acquisition of new functions was abruptly replaced by paralogization. We found homologs to herpesvirus surface glycoprotein gB in cytoplasmic viruses. This study provided the first prediction of an endonuclease in 10 of the 20 viruses examined; the first report in a virus of a phenolic acid decarboxylase, proteasomal subunit, or cysteine knot (defensin) protein; and the first report of a prokaryotic-type ribosomal protein in a eukaryotic virus.


Assuntos
Vírus de DNA/classificação , Vírus de DNA/genética , Vírus Gigantes/genética , Filogenia , Proteoma/genética , Proteínas Virais/genética , Anquirinas/genética , Citoplasma/virologia , RNA Polimerases Dirigidas por DNA , Células Eucarióticas , Evolução Molecular , Tamanho do Genoma , Genoma Viral , Mimiviridae/genética , Vaccinia/genética
19.
Front Immunol ; 11: 1458, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32765505

RESUMO

Modified Vaccinia virus Ankara (MVA) is an attenuated strain of vaccinia virus and currently under investigation as a promising vaccine vector against infectious diseases and cancer. MVA acquired mutations in host range and immunomodulatory genes, rendering the virus deficient for replication in most mammalian cells. MVA has a high safety profile and induces robust immune responses. However, the role of innate immune triggers for the induction of cytotoxic T cell responses after vaccination is incompletely understood. Stimulator of interferon genes (STING) is an adaptor protein which integrates signaling downstream of several DNA sensors and therefore mediates the induction of type I interferons and other cytokines or chemokines in response to various dsDNA viruses. Since the type I interferon response was entirely STING-dependent during MVA infection, we studied the effect of STING on primary and secondary cytotoxic T cell responses and memory T cell formation after MVA vaccination in STING KO mice. Moreover, we analyzed the impact of STING on the maturation of bone marrow-derived dendritic cells (BMDCs) and their functionality as antigen presenting cells for cytotoxic T cells during MVA infection in vitro. Our results show that STING has an impact on the antigen processing and presentation capacity of conventionel DCs and played a crucial role for DC maturation and type I interferon production. Importantly, STING was required for the induction of efficient cytotoxic T cell responses in vivo, since we observed significantly decreased short-lived effector and effector memory T cell responses after priming in STING KO mice. These findings indicate that STING probably integrates innate immune signaling downstream of different DNA sensors in DCs and shapes the cytotoxic T cell response via the DC maturation phenotype which strongly depends on type I interferons in this infection model. Understanding the detailed functions of innate immune triggers during MVA infection will contribute to the optimized design of MVA-based vaccines.


Assuntos
Células Dendríticas/imunologia , Vetores Genéticos/genética , Proteínas de Membrana/metabolismo , Linfócitos T Citotóxicos/imunologia , Vírus Vaccinia/genética , Vaccinia/imunologia , Animais , Apresentação do Antígeno , Células Cultivadas , Feminino , Humanos , Interferon Tipo I/metabolismo , Proteínas de Membrana/genética , Camundongos , Camundongos Knockout , Vacinação
20.
Viruses ; 12(8)2020 07 24.
Artigo em Inglês | MEDLINE | ID: mdl-32722032

RESUMO

The mass smallpox vaccination campaign has played a crucial role in smallpox eradication. Various strains of the vaccinia virus (VACV) were used as a live smallpox vaccine in different countries, their origin being unknown in most cases. The VACV strains differ in terms of pathogenicity exhibited upon inoculation of laboratory animals and reactogenicity exhibited upon vaccination of humans. Therefore, each generated strain or clonal variant of VACV needs to be thoroughly studied in in vivo systems. The clonal variant 14 of LIVP strain (LIVP-14) was the study object in this work. A comparative analysis of the virulence and immunogenicity of LIVP-14 inoculated intranasally (i.n.), intradermally (i.d.), or subcutaneously (s.c.) to BALB/c mice at doses of 108, 107, and 106 pfu was carried out. Adult mice exhibited the highest sensitivity to the i.n. administered LIVP-14 strain, although the infection was not lethal. The i.n. inoculated LIVP-14 replicated efficiently in the lungs. Furthermore, this virus was accumulated in the brain at relatively high concentrations. Significantly lower levels of LIVP-14 were detected in the liver, kidneys, and spleen of experimental animals. No clinical manifestations of the disease were observed after i.d. or s.c. injection of LIVP-14 to mice. After s.c. inoculation, the virus was detected only at the injection site, while it could disseminate to the liver and lungs when delivered via i.d. administration. A comparative analysis of the production of virus-specific antibodies by ELISA and PRNT revealed that the highest level of antibodies was induced in i.n. inoculated mice; a lower level of antibodies was observed after i.d. administration of the virus and the lowest level after s.c. injection. Even at the lowest studied dose (106 pfu), i.n. or i.d. administered LIVP-14 completely protected mice against infection with the cowpox virus at the lethal dose. Our findings imply that, according to the ratio between such characteristics as pathogenicity/immunogenicity/protectivity, i.d. injection is the optimal method of inoculation with the VACV LIVP-14 strain to ensure the safe formation of immune defense after vaccination against orthopoxviral infections.


Assuntos
Anticorpos Antivirais/sangue , Vírus Vaccinia/imunologia , Vírus Vaccinia/patogenicidade , Administração Intranasal , Animais , Anticorpos Neutralizantes/sangue , Vírus da Varíola Bovina/imunologia , Feminino , Imunogenicidade da Vacina , Injeções Intradérmicas , Injeções Subcutâneas , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Vacina Antivariólica , Vaccinia/prevenção & controle , Vaccinia/virologia , Vírus Vaccinia/classificação , Virulência
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