The highly conserved human leukocyte antigen-A2(HLA-A2)-restricted epitope NS3-1073 represents a promising candidate for a therapeutic vaccine against hepatitis C virus(HCV).In this study,we engineered a set of fusion...The highly conserved human leukocyte antigen-A2(HLA-A2)-restricted epitope NS3-1073 represents a promising candidate for a therapeutic vaccine against hepatitis C virus(HCV).In this study,we engineered a set of fusion proteins based on the artificial self-assembling peptide(SAP),which were expressed in Escherichia coli and spontaneously self-assembled into nanosized particles displaying HCV epitopes,including NS3-1073.To enhance immunogenicity,we incorporated the T helper epitope PADRE into the construct.Alpha-helical linkers were introduced between SAP and the epitopes to facilitate proper protein folding.Notably,a helical linker with a high supercoiling propensity enabled soluble expression of the fusion protein containing both the NS3-1073 and PADRE epitopes,allowing purification of the in vivo-formed nanoparticles by metal affinity chromatography.Human dendritic cells derived from peripheral blood monocytes showed robust activation in response to the fusion proteins and preferentially stimulated T lymphocytes toward a Th1-biased immune response.In mice,immunization with nanoparticles carrying NS3-1073 induced splenocyte proliferation in response to in vitro stimulation with a mixture of NS3 peptides.These results demonstrate that recombinant nanoparticle-based carriers presenting the NS3-1073 epitope can be produced in bacterial systems and hold strong potential as a foundation for a therapeutic HCV vaccine.展开更多
Dengue viruses(DENV)have spread throughout the world and pose a huge threat to human life.The most widespread serotype is type 2 DENV(DENV 2),which has no specific treatment.NS2B-NS3 protease plays a pivotal role in D...Dengue viruses(DENV)have spread throughout the world and pose a huge threat to human life.The most widespread serotype is type 2 DENV(DENV 2),which has no specific treatment.NS2B-NS3 protease plays a pivotal role in DENV replication because of its function in cleavage of the viral polyprotein;thus,it is considered a promising target for antiviral discovery.In this study,we developed a high-throughput screening system based on the NS2B-NS3 protease to identify candidates from an FDA-approved drug library.Eltrombopag was screened out of 3273 drugs,and demonstrated inhibition on DENV 2 at the micromolar level in vitro,significantly reducing viral loads in the targeted organs of challenged mice following intraperitoneal injection.Further mechanistic analysis showed that eltrombopag allosterically binds to the DENV 2 NS2B-NS3 protease in a reversible,noncompetitive manner,therefore inhibiting DENV 2 at the post-infection stage.In addition,eltrombopag inhibited the NS2B-NS3 proteases of DENV 4 and Zika virus,suggesting its potential as a broadspectrum antiviral agent.This study repurposed eltrombopag as a promising antiviral agent against DENV,providing an alternative for antiviral development against flaviviruses.展开更多
OBJECTIVE:To investigate the therapeutic effects of Jinxin oral liquid(金欣口服液,JX)on influenza A virus(H1N1)influenza virus infected mice.METHODS:We established a model of by intranasally infecting the mice with H1...OBJECTIVE:To investigate the therapeutic effects of Jinxin oral liquid(金欣口服液,JX)on influenza A virus(H1N1)influenza virus infected mice.METHODS:We established a model of by intranasally infecting the mice with H1N1 virus.The mice were then orally administered JX or ribavirin to evaluate their therapeutic effects in vivo.We conducted histologic and immunohistochemical analyses,enzyme linked immunosorbent assay or quantitative real-time polymerase chain reaction to assess lung damage and the expression of inflammatory cytokines.Western blot(WB)experiments was conducted to measure the activation of NOD-like receptor protein 3(NLRP3)pathway.Flow cytometry was employed to quantify the populations of alveolar macrophages(AMs).To block the NLRP3 pathway,mice were treated with MCC950.For AMs depletion,mice were intranasally administered a single dose of clodronate liposome.RESULTS:Administration of JX demonstrated a protective effect against H1N1-induced lung pathology by reducing lung injury,suppressing lung inflammation,and decreasing viral titer.JX significantly inhibited the production of pro-inflammatory cytokines,such as interleukin(IL)-1βand tumor necrosis factor-ɑ,in H1N1-infected mice.JX inhibits the activation of NOD-like receptor protein 3(NLRP3)/apoptosis-associated specklike protein containing a caspase recruitment domain/caspase 1 pathway in the lungs and AMs of H1N1-infected mice.The inhibitory effect of JX on IL-1βsecretion was mediated by blocking the NLRP3 pathway activation in AMs.CONCLUSIONS:These findings suggest that JX holds promise as a potential therapeutic agent for suppressing the aggressive pro-inflammatory response induced by H1N1 infection.Further research and development are warranted to explore the full potential of JX in the prevention and treatment of H1N1 infection。展开更多
BACKGROUND Hepatitis B virus(HBV)evades the innate immunity and leads to persistent chronic infection,but the molecular mechanism is still not well known.AIM To investigate whether HBV-miR-3 is involved in HBV immune ...BACKGROUND Hepatitis B virus(HBV)evades the innate immunity and leads to persistent chronic infection,but the molecular mechanism is still not well known.AIM To investigate whether HBV-miR-3 is involved in HBV immune evasion.METHODS HBV-miR-3 agomir and antagomir were employed to verify the effectiveness of HBV-miR-3 on cGAS-Sting-IFN pathway through the experiments on relative luciferase activity,cGAS protein expression,Sting phosphorylation and interferon(IFN)production.RESULTS HBV-miR-3 down-regulates cGAS protein expression post-transcriptionally by inhibition of cGAS 3’-untranslated region(3’-UTR)activity,which results in lower Sting phosphorylation and IFN production.HBV-miR-3 antagomir rescued cGAS protein expression,Sting phosphorylation and IFN-βproduction.CONCLUSION HBV-miR-3 plays an important role in HBV immunity evasion by targeting cGAS 3’-UTR and interfering with cGAS-Sting-IFN pathway.展开更多
【目的】人副流感病毒3型(human parainfluenza virus type 3,HPIV-3)是全球获得性呼吸道感染的关键致病因素,目前尚无特异性治疗药物。鉴于该病原体抗原具有复杂性和变异性,其疫苗开发进展较为滞后。设计具有广谱特性的新型疫苗,对于...【目的】人副流感病毒3型(human parainfluenza virus type 3,HPIV-3)是全球获得性呼吸道感染的关键致病因素,目前尚无特异性治疗药物。鉴于该病原体抗原具有复杂性和变异性,其疫苗开发进展较为滞后。设计具有广谱特性的新型疫苗,对于为持续突变的野生型毒株提供全面防护至关重要。【方法】为克服病毒的抗原性变异,从NCBI下载不同的HPIV-3抗原蛋白(F、M、N和HN蛋白)序列,并通过序列比对分别生成共识序列,进一步运用反向疫苗学方法,预测并设计了一种针对HPIV-3的广谱T细胞表位疫苗。【结果】该多表位疫苗包含来自F、M、N和HN蛋白的11个CTL表位(9-mer)和11个HTL表位(15-mer),由355个氨基酸组成,不含佐剂。预测的T细胞表位具备可溶性、非致敏性、高抗原性及免疫原性。设计的疫苗能有效结合天然免疫中的Toll样受体,具有较好的稳定性和亲水性,且人口覆盖度较高。【结论】本研究设计的HPIV-3多表位疫苗有望成为预防HPIV-3感染的候选疫苗,为疫苗的设计与开发提供了一种新颖的免疫信息学方法。展开更多
基金supported by the Russian Science Foundation(Grant No.24-25-20087 to V.K.)。
文摘The highly conserved human leukocyte antigen-A2(HLA-A2)-restricted epitope NS3-1073 represents a promising candidate for a therapeutic vaccine against hepatitis C virus(HCV).In this study,we engineered a set of fusion proteins based on the artificial self-assembling peptide(SAP),which were expressed in Escherichia coli and spontaneously self-assembled into nanosized particles displaying HCV epitopes,including NS3-1073.To enhance immunogenicity,we incorporated the T helper epitope PADRE into the construct.Alpha-helical linkers were introduced between SAP and the epitopes to facilitate proper protein folding.Notably,a helical linker with a high supercoiling propensity enabled soluble expression of the fusion protein containing both the NS3-1073 and PADRE epitopes,allowing purification of the in vivo-formed nanoparticles by metal affinity chromatography.Human dendritic cells derived from peripheral blood monocytes showed robust activation in response to the fusion proteins and preferentially stimulated T lymphocytes toward a Th1-biased immune response.In mice,immunization with nanoparticles carrying NS3-1073 induced splenocyte proliferation in response to in vitro stimulation with a mixture of NS3 peptides.These results demonstrate that recombinant nanoparticle-based carriers presenting the NS3-1073 epitope can be produced in bacterial systems and hold strong potential as a foundation for a therapeutic HCV vaccine.
基金supported by the National Natural Science Foundation of China(Grant No.82130101)the Youth Innovation Promotion Association of CAS(Grant No.2021333).
文摘Dengue viruses(DENV)have spread throughout the world and pose a huge threat to human life.The most widespread serotype is type 2 DENV(DENV 2),which has no specific treatment.NS2B-NS3 protease plays a pivotal role in DENV replication because of its function in cleavage of the viral polyprotein;thus,it is considered a promising target for antiviral discovery.In this study,we developed a high-throughput screening system based on the NS2B-NS3 protease to identify candidates from an FDA-approved drug library.Eltrombopag was screened out of 3273 drugs,and demonstrated inhibition on DENV 2 at the micromolar level in vitro,significantly reducing viral loads in the targeted organs of challenged mice following intraperitoneal injection.Further mechanistic analysis showed that eltrombopag allosterically binds to the DENV 2 NS2B-NS3 protease in a reversible,noncompetitive manner,therefore inhibiting DENV 2 at the post-infection stage.In addition,eltrombopag inhibited the NS2B-NS3 proteases of DENV 4 and Zika virus,suggesting its potential as a broadspectrum antiviral agent.This study repurposed eltrombopag as a promising antiviral agent against DENV,providing an alternative for antiviral development against flaviviruses.
基金Young Elite Scientists Sponsorship Program by Chinese Association for Clinical Medicine(2021-QNRC2-B14)Colleges and universities in Jiangsu Province Natural Science Research:Investigating the Mechanism of Jinping Decoction Inhibiting the Differentiation of Myeloid-derived Suppressor Cells and Regulating Lipid Metabolism(22KJA360004)+3 种基金Jiangsu Graduate Practice Innovation Program:A Study on the Mechanism of Xiaofeng Xuanqiao Decoction in Treating Pediatric Rhinitis based on Interleukin-33/Suppression of Tumorigenicity 2(SJCX23_0803)Wuxi Health Commission Scientific Research Project:Observation on the Efficacy of Xiaofeng Xuanqiao Decoction in Treating Pediatric Rhinitis(WindPhlegm Obstructing Orifices Syndrome)and Its Influence on Serum IgE and Related Inflammatory Cytokine Levels(M202035)Project of High-level Construction of Key Traditional Chinese Medicine(TCM)Disciplines,China:Research on Tic Disorders from the Perspective of the Five Viscera Syndrome Treatment(NZYEK004)Project of Highlevel Construction of Key TCM Disciplines,China:Research on the Mechanism of Xiaofeng Xuanqiao Decoction in Treating Allergic Rhinitis based on the Interleukin-33/ST2 Axis(NZYEK006)。
文摘OBJECTIVE:To investigate the therapeutic effects of Jinxin oral liquid(金欣口服液,JX)on influenza A virus(H1N1)influenza virus infected mice.METHODS:We established a model of by intranasally infecting the mice with H1N1 virus.The mice were then orally administered JX or ribavirin to evaluate their therapeutic effects in vivo.We conducted histologic and immunohistochemical analyses,enzyme linked immunosorbent assay or quantitative real-time polymerase chain reaction to assess lung damage and the expression of inflammatory cytokines.Western blot(WB)experiments was conducted to measure the activation of NOD-like receptor protein 3(NLRP3)pathway.Flow cytometry was employed to quantify the populations of alveolar macrophages(AMs).To block the NLRP3 pathway,mice were treated with MCC950.For AMs depletion,mice were intranasally administered a single dose of clodronate liposome.RESULTS:Administration of JX demonstrated a protective effect against H1N1-induced lung pathology by reducing lung injury,suppressing lung inflammation,and decreasing viral titer.JX significantly inhibited the production of pro-inflammatory cytokines,such as interleukin(IL)-1βand tumor necrosis factor-ɑ,in H1N1-infected mice.JX inhibits the activation of NOD-like receptor protein 3(NLRP3)/apoptosis-associated specklike protein containing a caspase recruitment domain/caspase 1 pathway in the lungs and AMs of H1N1-infected mice.The inhibitory effect of JX on IL-1βsecretion was mediated by blocking the NLRP3 pathway activation in AMs.CONCLUSIONS:These findings suggest that JX holds promise as a potential therapeutic agent for suppressing the aggressive pro-inflammatory response induced by H1N1 infection.Further research and development are warranted to explore the full potential of JX in the prevention and treatment of H1N1 infection。
基金Supported by National Natural Science Foundation of China,Key Project,No.82430071The Scientific Research Program of FuRong Laboratory,No.2023SK2108+2 种基金Clinical Medical Research Center for Viral Hepatitis of Hunan Province,No.2023SK4009Hunan Provincial Natural Science Foundation,No.2023JJ60440Hunan Provincial Health Commission Research Program,No.C202303088786.
文摘BACKGROUND Hepatitis B virus(HBV)evades the innate immunity and leads to persistent chronic infection,but the molecular mechanism is still not well known.AIM To investigate whether HBV-miR-3 is involved in HBV immune evasion.METHODS HBV-miR-3 agomir and antagomir were employed to verify the effectiveness of HBV-miR-3 on cGAS-Sting-IFN pathway through the experiments on relative luciferase activity,cGAS protein expression,Sting phosphorylation and interferon(IFN)production.RESULTS HBV-miR-3 down-regulates cGAS protein expression post-transcriptionally by inhibition of cGAS 3’-untranslated region(3’-UTR)activity,which results in lower Sting phosphorylation and IFN production.HBV-miR-3 antagomir rescued cGAS protein expression,Sting phosphorylation and IFN-βproduction.CONCLUSION HBV-miR-3 plays an important role in HBV immunity evasion by targeting cGAS 3’-UTR and interfering with cGAS-Sting-IFN pathway.
文摘【目的】人副流感病毒3型(human parainfluenza virus type 3,HPIV-3)是全球获得性呼吸道感染的关键致病因素,目前尚无特异性治疗药物。鉴于该病原体抗原具有复杂性和变异性,其疫苗开发进展较为滞后。设计具有广谱特性的新型疫苗,对于为持续突变的野生型毒株提供全面防护至关重要。【方法】为克服病毒的抗原性变异,从NCBI下载不同的HPIV-3抗原蛋白(F、M、N和HN蛋白)序列,并通过序列比对分别生成共识序列,进一步运用反向疫苗学方法,预测并设计了一种针对HPIV-3的广谱T细胞表位疫苗。【结果】该多表位疫苗包含来自F、M、N和HN蛋白的11个CTL表位(9-mer)和11个HTL表位(15-mer),由355个氨基酸组成,不含佐剂。预测的T细胞表位具备可溶性、非致敏性、高抗原性及免疫原性。设计的疫苗能有效结合天然免疫中的Toll样受体,具有较好的稳定性和亲水性,且人口覆盖度较高。【结论】本研究设计的HPIV-3多表位疫苗有望成为预防HPIV-3感染的候选疫苗,为疫苗的设计与开发提供了一种新颖的免疫信息学方法。
