A5066029443- Mosquito-borne diseases and control
- Viral Infections and Vectors
- HIV Research and Treatment
- Immunotherapy and Immune Responses
- Viral Infections and Outbreaks Research
- vaccines and immunoinformatics approaches
- Virology and Viral Diseases
- Viral gastroenteritis research and epidemiology
- Hepatitis B Virus Studies
- Virus-based gene therapy research
- Immune Cell Function and Interaction
- Bacteriophages and microbial interactions
- interferon and immune responses
- Animal Disease Management and Epidemiology
- Immune Response and Inflammation
- Herpesvirus Infections and Treatments
- Bacterial Genetics and Biotechnology
- Respiratory viral infections research
- RNA Interference and Gene Delivery
- Malaria Research and Control
- Hepatitis C virus research
- T-cell and B-cell Immunology
- RNA and protein synthesis mechanisms
- SARS-CoV-2 and COVID-19 Research
- CRISPR and Genetic Engineering
Karolinska Institutet
2012-2021
University of Oxford
2009-2012
Swedish Institute
1999-2011
University of Helsinki
1981-2009
Cancer Research UK
2006
University of Freiburg
2006
Robert Koch Institute
2004
Trinity College Dublin
1996
Colorado State University
1996
Washington University in St. Louis
1995
Double-stranded RNA (dsRNA) produced during viral replication is believed to be the critical trigger for activation of antiviral immunity mediated by helicase enzymes retinoic acid–inducible gene I (RIG-I) and melanoma differentiation–associated 5 (MDA5). We showed that influenza A virus infection does not generate dsRNA RIG-I activated genomic single-stranded (ssRNA) bearing 5′-phosphates. This blocked protein nonstructured 1 (NS1), which found in a complex with infected cells. These...
We report on the construction of a full-length cDNA clone Semliki Forest virus (SFV). By placing under SP6 promoter, infectious RNA can be produced in vitro and used to transfect cells initiate infection. To achieve efficient transfections, new protocol for electroporation was developed. This method gave up 500-fold improvement over traditional DEAE-dextran transfection procedure. Since virtually 100% transfected by electroporation, this is useful tool detailed biochemical studies null...
The EuroVacc 02 phase I trial has evaluated the safety and immunogenicity of a prime-boost regimen comprising recombinant DNA poxvirus vector NYVAC, both expressing common immunogen consisting Env, Gag, Pol, Nef polypeptide domain from human immunodeficiency virus (HIV)-1 clade C isolate, CN54. 40 volunteers were randomized to receive or nothing on day 0 at week 4, followed by NYVAC weeks 20 24. primary endpoints measured 26 28 quantification T cell responses using interferon γ enzyme-linked...
Despite impressive clinical success, cancer immunotherapy based on immune checkpoint blockade remains ineffective in many patients due to tumoral resistance. Here we use the autochthonous TiRP melanoma model, which recapitulates resistance signature observed human melanomas. tumors resist blockade, vaccines or adoptive T-cell therapy. recruit and activate tumor-specific CD8+ T cells, but these cells then undergo apoptosis. This does not occur with isogenic transplanted tumors, are rejected...
Alphavirus infection results in the shutoff of host protein synthesis favor viral translation. Here, we show that during Semliki Forest virus (SFV) infection, translation inhibition is largely due to activation cellular stress response via phosphorylation eukaryotic initiation factor 2alpha subunit (eIF2alpha). Infection mouse embryo fibroblasts (MEFs) expressing a nonphosphorylatable mutant eIF2alpha does not result efficient shutoff, despite production. Furthermore, SFV enhancer element...
Alphavirus expression systems based on suicidal virus particles carrying recombinant replicons have proven to be a very efficient way deliver genes for heterologous protein expression. However, present strategies production of such biosafety limitations due the generation, by RNA recombination, replication-proficient viruses (RPVs). Here we describe new packaging system Semliki Forest (SFV) use two-helper in which capsid and spike proteins C-p62-6K-E1 polyprotein are expressed from two...
The proteolytic processes involved in the cotranslational production of Semliki Forest virus proteins p62, 6K, and E1 from a common precursor polypeptide were analyzed by an vitro translation-translocation assay. By studying behavior wild-type mutant variants polyprotein, we show that signal sequences responsible for membrane translocation 6K reside C-terminal regions p62 respectively. We present evidence suggesting polyprotein is processed on luminal side peptidase at consensus cleavage...
ABSTRACT Chikungunya virus (CHIKV) is a reemerging mosquito-borne alphavirus that has caused severe epidemics in Africa and Asia occasionally Europe. As of today, there no licensed vaccine available to prevent CHIKV infection. Here we describe the development evaluation novel candidates were attenuated by deleting large part gene encoding nsP3 or entire 6K administered as viral particles infectious genomes launched DNA. The resulting mutants genetically stable elicited high magnitudes...
Semliki Forest virus (SFV) particles are released from infected cells by budding of nucleocapsids through plasma membrane regions that modified spike proteins. The process was studied with recombinant SFV genomes which lacked the nucleocapsid protein gene or, alternatively, genes. No subviral were expressed only or Virus release found to be strictly dependent on coexpression and These results provide direct proof for hypothesis alphavirus is driven nucleocapsid-spike interactions. importance...
There is a need to develop single and highly effective vaccine against the emerging chikungunya virus (CHIKV), which causes severe disease in humans. Here, we have generated characterized immunogenicity profile efficacy of novel CHIKV candidate based on attenuated poxvirus vector modified vaccinia Ankara (MVA) expressing C, E3, E2, 6K, E1 structural genes (termed MVA-CHIKV). MVA-CHIKV was stable cell culture, expressed proteins, triggered cytoplasmic accumulation Golgi apparatus-derived...
Chikungunya virus (CHIKV) is rapidly spreading across the globe, and millions are infected. Morbidity due to this a serious threat public health, but at present, there no vaccine against debilitating disease. We have recently developed number of candidates, here we evaluated 3 them in nonhuman primate model. A single immunization with an attenuated strain CHIKV (Δ5nsP3), homologous prime-boost DNA-launched RNA replicon encoding envelope proteins (DREP-E), DREP-E prime followed by recombinant...
Abstract Recombinant vaccines encoding strings of virus- or tumor-derived peptides and/or proteins are currently being designed for use against both cancer and infectious diseases. These aim to induce cytotoxic immune responses several Ags simultaneously. We developed a novel tetramer-based technique, based on chimeric HLA A2/H-2Kb H chains, directly monitor the CTL response such in HLA-A2 transgenic mice. found that priming boosting with same polyepitope construct induced were dominated by...
The envelope of the Semliki Forest virus (SFV) contains two transmembrane proteins, E2 and E1, in a heterodimeric complex. subunit is initially synthesized as precursor protein p62, which proteolytically processed to mature form before budding at plasma membrane. p62 (E2) mediates binding heterodimer nucleocapsid during budding, whereas E1 carries entry functions virus, that is, cell low pH-mediated membrane fusion activity. We have investigated significance cleavage event for maturation...
Infection of pigtail macaques with SIVsmmPBj14, biological clone 3 (SIV-PBj14-bc13), produces an acute and usually fatal shock-like syndrome 7 to 14 days after infection. We used this simian immunodeficiency virus (SIV) model as a rapid rigorous challenge evaluate the efficacy two SIV Env vaccine strategies. Groups four were immunized times over 25-week span either recombinant Semliki Forest expressing SIV-PBj14 gp160 (SFV-SIVgp160) or purified gp120 (rgp120) in SBN-1 adjuvant. Antibody...
In genetic vaccination, recipients are immunized with antigen-encoding nucleic acid, usually DNA. This study addressed the possibility of using recombinant alpha virus RNA molecule, which replicates in cytoplasm transfected cells, as a novel approach for vaccination. Mice were Semliki Forest RNA–encoding envelope proteins from one 3 viruses: influenza A virus, tickborne flavivirus (louping ill virus), or respiratory syncytial (RSV). Serologic analyses showed that antigen-specific antibody...
About 50% of Semliki Forest virus-specific nonstructural protein nsP2 is associated with the nuclear fraction in virus-infected BHK cells. Transport into nucleus must be specific, since only trace amounts nsP3 and nsP4 about 13% nsP1, all derived from same polyprotein, were found nucleus. Subfractionation [35S]methionine-labeled cells showed that 80 to 90% was matrix. Indirect immunofluorescence, anti-nsP2 antiserum, most intensive staining structures which by Nomarski optics appeared...
Infection of macaques with chimeric simian-human immunodeficiency viruses (SHIVs) allows evaluation HIV-1 envelope vaccines. SHIV-4 is based on SIVmac239 but carries the env, tat, and rev genes HIV-1IIIB. In this study we used Semliki Forest virus (SFV) RNA vectors to express protein gp160 HIV-1IIIB in cynomolgus macaques. Monkeys were immunized four times recombinant suicide SFV. Whereas two monkeys showed T cell-proliferative responses, only one monkey had demonstrable levels antibodies...