Horseshoe bats host numerous SARS-related coronaviruses without overt disease signs.Bat intestinal organoids,a unique model of bat intestinal epithelium,allow direct comparison with human intestinal organoids.We sough...Horseshoe bats host numerous SARS-related coronaviruses without overt disease signs.Bat intestinal organoids,a unique model of bat intestinal epithelium,allow direct comparison with human intestinal organoids.We sought to unravel the cellular mechanism(s)underlying bat tolerance of coronaviruses by comparing the innate immunity in bat and human organoids.We optimized the culture medium,which enabled a consecutive passage of bat intestinal organoids for over one year.Basal expression levels of IFNs and IFN-stimulated genes were higher in bat organoids than in their human counterparts.Notably,bat organoids mounted a more rapid,robust and prolonged antiviral defense than human organoids upon Poly(I:C)stimulation.TLR3 and RLR might be the conserved pathways mediating antiviral response in bat and human intestinal organoids.The susceptibility of bat organoids to a bat coronavirus CoV-HKU4,but resistance to EV-71,an enterovirus of exclusive human origin,indicated that bat organoids adequately recapitulated the authentic susceptibility of bats to certain viruses.Importantly,TLR3/RLR inhibition in bat organoids significantly boosted viral growth in the early phase after SARS-CoV-2 or CoV-HKU4 infection.Collectively,the higher basal expression of antiviral genes,especially more rapid and robust induction of innate immune response,empowered bat cells to curtail virus propagation in the early phase of infection.展开更多
We conducted a six-year epidemiological study on human coronaviruses(HCoVs) circulating in Hong Kong, using 8275 nasopharyngeal samples from patients with acute respiratory tract infections. HCoVs were detected in 77(...We conducted a six-year epidemiological study on human coronaviruses(HCoVs) circulating in Hong Kong, using 8275 nasopharyngeal samples from patients with acute respiratory tract infections. HCoVs were detected in 77(0.93%) of the samples by a pan-HCoV RT-PCR assay. The most frequently detected HCoV species was HCoV-OC43(0.58%), followed by HCoV-229E(0.15%),HCoV-HKU1(0.13%) and HCoV-NL63(0.07%). HCoVs were detected throughout the study period(September 2008–August 2014), with the highest detection rate from September 2010 to August2011(22/1500, 1.47%). Different seasonal patterns of each HCoV species in Hong Kong were noted.HCoV-OC43 was predominant in the fall and winter, whereas HCoV-HKU1 showed peak activity in winter, with a few cases occurred in spring and summer. HCoV-229 E mainly occurred in winter and spring, while HCoV-NL63 was predominant in summer and autumn. HCoVs most commonly infect the elderly and young children, with median age of 79.5 years(range, 22 days to 95 years).Intriguingly, the detection rate of HCoV-OC43 in the age group of > 80 years(26/2380, 1.09%) was significantly higher than that in the age group of 0–10 years(12/2529, 0.47%)(P < 0.05). These data provides new insight into the epidemiology of coronaviruses.展开更多
Historically,influenza pandemics have arisen from avian influenza viruses.Avian influenza viruses H5N1 and H9N2 are potential pandemic candidates.Infection of humans with the highly pathogenic avian influenza H5N1 vir...Historically,influenza pandemics have arisen from avian influenza viruses.Avian influenza viruses H5N1 and H9N2 are potential pandemic candidates.Infection of humans with the highly pathogenic avian influenza H5N1 virus is associated with a mortality in excess of 60%,which has been attributed to dysregulation of the cytokine system.Human macrophages and epithelial cells infected with some genotypes of H5N1 and H9N2 viruses express markedly elevated cytokine and chemokine levels when compared with seasonal influenza A subtype H1N1 virus.The mechanisms underlying this cytokine and chemokine hyperinduction are not fully elucidated.In the present study,we demonstrate that autophagy,a tightly regulated homeostatic process for self-digestion of unwanted cellular subcomponents,plays a role in cytokine induction.Autophagy is induced to a greater extent by H9N2/G1,in association with cytokine hyperinduction,compared with H1N1 and the novel pandemic swine-origin influenza A/H1N1 viruses.Using 3-methyladenine to inhibit autophagy and small interfering RNA to silence the autophagy gene,Atg5,we further show that autophagic responses play a role in influenza virus-induced CXCL10 and interferon-a expression in primary human blood macrophages.Our results provide new insights into the pathogenic mechanisms of avian influenza viruses.展开更多
Live attenuated vaccines might elicit mucosal and sterilizing immunity against SARS-CoV-2 that the existing mRNA,adenoviral vector and inactivated vaccines fail to induce.Here,we describe a candidate live attenuated v...Live attenuated vaccines might elicit mucosal and sterilizing immunity against SARS-CoV-2 that the existing mRNA,adenoviral vector and inactivated vaccines fail to induce.Here,we describe a candidate live attenuated vaccine strain of SARS-CoV-2 in which the NSP16 gene,which encodes 2′-O-methyltransferase,is catalytically disrupted by a point mutation.This virus,designated d16,was severely attenuated in hamsters and transgenic mice,causing only asymptomatic and nonpathogenic infection.A single dose of d16 administered intranasally resulted in sterilizing immunity in both the upper and lower respiratory tracts of hamsters,thus preventing viral spread in a contact-based transmission model.It also robustly stimulated humoral and cell-mediated immune responses,thus conferring full protection against lethal challenge with SARS-CoV-2 in a transgenic mouse model.The neutralizing antibodies elicited by d16 effectively cross-reacted with several SARS-CoV-2 variants.Secretory immunoglobulin A was detected in the blood and nasal wash of vaccinated mice.Our work provides proof-of-principle evidence for harnessing NSP16-deficient SARS-CoV-2 for the development of live attenuated vaccines and paves the way for further preclinical studies of d16 as a prototypic vaccine strain,to which new features might be introduced to improve safety,transmissibility,immunogenicity and efficacy.展开更多
A straight, non-sporulating, Gram-variable bacillus (HKU24T) was recovered from the blood culture of a patient with metastatic breast carcinoma. After repeated subculturing in BACTEC Plus Anaerobic/F blood culture bro...A straight, non-sporulating, Gram-variable bacillus (HKU24T) was recovered from the blood culture of a patient with metastatic breast carcinoma. After repeated subculturing in BACTEC Plus Anaerobic/F blood culture broth, HKU24T grew on brucella agar as non-hemolytic, pinpoint colonies after 96 h of incubation at 37 °C in an anaerobic environment and aerobic environment with 5% CO2. Growth was enhanced with a streak of Staphylococcus aureus. HKU24T was non-motile and catalase-negative, but positive for alkaline phosphatase, β-glucosidase, and α-glucosidase. It hydrolyzed phenylphosphonate and reduced resazurin. 16S rRNA, groEL, gyrB, recA, and rpoB sequencing showed that HKU24T occupies a distinct phylogenetic position among the Leptotrichia species, being most closely related to Leptotrichia trevisanii. Using HKU24T groEL, gyrB, recA, and rpoB gene-specific primers, fragments of these genes were amplified from one of 20 oral specimens. Based on phenotypic and genotypic characteristics, we propose a new species, Leptotrichia hongkongensis sp. nov., to describe this bacterium.展开更多
Bats are connected with the increasing numbers of emerging and re-emerging viruses that may break the species barrier and spread into the human population. Coronaviruses are one of the most common viruses discovered i...Bats are connected with the increasing numbers of emerging and re-emerging viruses that may break the species barrier and spread into the human population. Coronaviruses are one of the most common viruses discovered in bats, which were considered as the natural source of recent human-susceptible coronaviruses, i.e. SARS-COV and MERS-CoV. Our previous study reported the discovery of a bat-derived putative cross-family recombinant coronavirus with a reovirus gene p10, named as Ro-BatCoV GCCDC1. In this report, through a two-year follow-up of a special bat population in one specific cave of south China, we illustrate that Ro-BatCoV GCCDC1 persistently circulates among bats. Notably, through the longitudinal observation, we identified the dynamic evolution of Ro-BatCoV GCCDC1 in bats represented by continuously recombination events. Our study provides the first glimpse of the virus evolution in one longitudinally observed bat population cohort and underlines the surveillance and pre-warning of potential interspecies transmittable viruses in bats.展开更多
Avian influenza viruses have been first known to jump the interspecies barrier and cause human infections since 1959[1].However,avian influenza was not considered a serious threat to public health until 1997,when the ...Avian influenza viruses have been first known to jump the interspecies barrier and cause human infections since 1959[1].However,avian influenza was not considered a serious threat to public health until 1997,when the influenza A(H5N1)virus caused an outbreak in Hong Kong with 18 cases including six deaths[2].The A(H5N1)epidemic has never stopped despite poultry immu-展开更多
The pandemic H1N12009 influenza virus has caused thefirst influenza pandemic of the 21st century,leading to disproportionate fatalities in the low-risk population despite the generally mild nature of the illness.Advances...The pandemic H1N12009 influenza virus has caused thefirst influenza pandemic of the 21st century,leading to disproportionate fatalities in the low-risk population despite the generally mild nature of the illness.Advances in science and technology have allowed very detailed study on the pathogenesis of this novel virus,and many have already been published in less than a year after the start of the pandemic.Information generated from cell lines,animal models,and clinical data analysis has provided us with greater understanding of the behavior of this virus and the associated host response.The new knowledge will allow us to formulate scientifically sound and evidence-based management plans.展开更多
The emergence of SARS-CoV-2 variants of concern and repeated outbreaks of coronavirus epidemics in the past two decades emphasize the need for next-generation pan-coronaviral therapeutics.Drugging the multi-functional...The emergence of SARS-CoV-2 variants of concern and repeated outbreaks of coronavirus epidemics in the past two decades emphasize the need for next-generation pan-coronaviral therapeutics.Drugging the multi-functional papain-like protease(PLpro)domain of the viral nsp3 holds promise.However,none of the known coronavirus PLpro inhibitors has been shown to be in vivo active.Herein,we screened a structurally diverse library of 50,080 compounds for potential coronavirus PLpro inhibitors and identified a noncovalent lead inhibitor F0213 that has broad-spectrum anti-coronaviral activity,including against the Sarbecoviruses(SARSCoV-1 and SARS-CoV-2),Merbecovirus(MERS-CoV),as well as the Alphacoronavirus(hCoV-229E and hCoVOC43).Importantly,F0213 confers protection in both SARS-CoV-2-infected hamsters and MERS-CoV-infected human DPP4-knockin mice.F0213 possesses a dual therapeutic functionality that suppresses coronavirus replication via blocking viral polyprotein cleavage,as well as promoting antiviral immunity by antagonizing the PLpro deubiquitinase activity.Despite the significant difference of substrate recognition,mode of inhibition studies suggest that F0213 is a competitive inhibitor against SARS2-PLpro via binding with the 157K amino acid residue,whereas an allosteric inhibitor of MERSPLpro interacting with its 271E position.Our proof-ofconcept findings demonstrated that PLpro is a valid target for the development of broad-spectrum anticoronavirus agents.The orally administered F0213 may serve as a promising lead compound for combating the ongoing COVID-19 pandemic and future coronavirus outbreaks.展开更多
Defective interfering genes(DIGs)are short viral genomes and interfere with wild-type viral replication.Here,we demonstrate that the new designed SARS-CoV-2 DIG(CD3600)can significantly inhibit the replication of SARS...Defective interfering genes(DIGs)are short viral genomes and interfere with wild-type viral replication.Here,we demonstrate that the new designed SARS-CoV-2 DIG(CD3600)can significantly inhibit the replication of SARS-CoV-2 including Alpha,Delta,Kappa and Omicron variants in human HK-2 cells and influenza DIG(PAD4)can significantly inhibit influenza virus replication in human A549 cells.One dose of influenza DIGs prophylactically protects 90%mice from lethal challenge of A(H1N1)pdm09 virus and CD3600 inhibits SARS-CoV-2 replication in hamster lungs when DIGs are administrated to lungs one day before viral challenge.To further investigate the gene delivery vector in the respiratory tract,a peptidic TAT2-P1&LAH4,which can package genes to form small spherical nanoparticles with high endosomal escape ability,is demonstrated to dramatically increase gene expression in the lung airway.TAT2-P1&LAH4,with the dual-functional TAT2-P1(gene-delivery and antiviral),can deliver CD3600 to significantly inhibit the replication of Delta and Omicron SARS-CoV-2 in hamster lungs.This peptide-based nanoparticle system can effectively transfect genes in lungs and deliver DIGs to inhibit SARS-CoV-2 variants and influenza virus in vivo,which provides the new insight into the drug delivery system for gene therapy against respiratory viruses.展开更多
基金supported by funding from the Health and Medical Research Fund(HMRF,17161272 and 19180392)of the Food and Health Bureau of the HKSAR government to J.Z.General Research Fund(GRF,17105420)+1 种基金Collaborative Research Fund(CRF,C7042-21G)Theme-based Research Scheme(TbRS,T11-709/21-N)of the Research Grants Council of HKSAR government to J.Z.,Health@InnoHK,Innovation and Technology Commission,HKSAR Government to K.Y.Y.
文摘Horseshoe bats host numerous SARS-related coronaviruses without overt disease signs.Bat intestinal organoids,a unique model of bat intestinal epithelium,allow direct comparison with human intestinal organoids.We sought to unravel the cellular mechanism(s)underlying bat tolerance of coronaviruses by comparing the innate immunity in bat and human organoids.We optimized the culture medium,which enabled a consecutive passage of bat intestinal organoids for over one year.Basal expression levels of IFNs and IFN-stimulated genes were higher in bat organoids than in their human counterparts.Notably,bat organoids mounted a more rapid,robust and prolonged antiviral defense than human organoids upon Poly(I:C)stimulation.TLR3 and RLR might be the conserved pathways mediating antiviral response in bat and human intestinal organoids.The susceptibility of bat organoids to a bat coronavirus CoV-HKU4,but resistance to EV-71,an enterovirus of exclusive human origin,indicated that bat organoids adequately recapitulated the authentic susceptibility of bats to certain viruses.Importantly,TLR3/RLR inhibition in bat organoids significantly boosted viral growth in the early phase after SARS-CoV-2 or CoV-HKU4 infection.Collectively,the higher basal expression of antiviral genes,especially more rapid and robust induction of innate immune response,empowered bat cells to curtail virus propagation in the early phase of infection.
基金partly supported by the Theme-Based Research Scheme, University Grant CouncilStrategic Research Theme Fund and University Development Fund, The University of Hong KongHealth and Medical Research Fund of the Food and Health Bureau of HKSAR
文摘We conducted a six-year epidemiological study on human coronaviruses(HCoVs) circulating in Hong Kong, using 8275 nasopharyngeal samples from patients with acute respiratory tract infections. HCoVs were detected in 77(0.93%) of the samples by a pan-HCoV RT-PCR assay. The most frequently detected HCoV species was HCoV-OC43(0.58%), followed by HCoV-229E(0.15%),HCoV-HKU1(0.13%) and HCoV-NL63(0.07%). HCoVs were detected throughout the study period(September 2008–August 2014), with the highest detection rate from September 2010 to August2011(22/1500, 1.47%). Different seasonal patterns of each HCoV species in Hong Kong were noted.HCoV-OC43 was predominant in the fall and winter, whereas HCoV-HKU1 showed peak activity in winter, with a few cases occurred in spring and summer. HCoV-229 E mainly occurred in winter and spring, while HCoV-NL63 was predominant in summer and autumn. HCoVs most commonly infect the elderly and young children, with median age of 79.5 years(range, 22 days to 95 years).Intriguingly, the detection rate of HCoV-OC43 in the age group of > 80 years(26/2380, 1.09%) was significantly higher than that in the age group of 0–10 years(12/2529, 0.47%)(P < 0.05). These data provides new insight into the epidemiology of coronaviruses.
基金supported by Area of Excellence grants to Malik Peiris,K.Y.Yuen and Allan S.Lau(Grant AoE/M-12/06)from the Research Grants Council of Hong Kong and the Research Fund for Control of Infectious Disease(09080832),as well as grants to Allan S.Lau and Malik Peiris from the Research Grants Council Central Allocation(HKU 1/05C).
文摘Historically,influenza pandemics have arisen from avian influenza viruses.Avian influenza viruses H5N1 and H9N2 are potential pandemic candidates.Infection of humans with the highly pathogenic avian influenza H5N1 virus is associated with a mortality in excess of 60%,which has been attributed to dysregulation of the cytokine system.Human macrophages and epithelial cells infected with some genotypes of H5N1 and H9N2 viruses express markedly elevated cytokine and chemokine levels when compared with seasonal influenza A subtype H1N1 virus.The mechanisms underlying this cytokine and chemokine hyperinduction are not fully elucidated.In the present study,we demonstrate that autophagy,a tightly regulated homeostatic process for self-digestion of unwanted cellular subcomponents,plays a role in cytokine induction.Autophagy is induced to a greater extent by H9N2/G1,in association with cytokine hyperinduction,compared with H1N1 and the novel pandemic swine-origin influenza A/H1N1 viruses.Using 3-methyladenine to inhibit autophagy and small interfering RNA to silence the autophagy gene,Atg5,we further show that autophagic responses play a role in influenza virus-induced CXCL10 and interferon-a expression in primary human blood macrophages.Our results provide new insights into the pathogenic mechanisms of avian influenza viruses.
基金supported by the Hong Kong Health and Medical Research Fund grants COVID190121 to JF-WC and COVID190114 to D-YJthe Hong Kong Research Grants Council grants C7142-20GF and T11-709/21-N to D-YJ.
文摘Live attenuated vaccines might elicit mucosal and sterilizing immunity against SARS-CoV-2 that the existing mRNA,adenoviral vector and inactivated vaccines fail to induce.Here,we describe a candidate live attenuated vaccine strain of SARS-CoV-2 in which the NSP16 gene,which encodes 2′-O-methyltransferase,is catalytically disrupted by a point mutation.This virus,designated d16,was severely attenuated in hamsters and transgenic mice,causing only asymptomatic and nonpathogenic infection.A single dose of d16 administered intranasally resulted in sterilizing immunity in both the upper and lower respiratory tracts of hamsters,thus preventing viral spread in a contact-based transmission model.It also robustly stimulated humoral and cell-mediated immune responses,thus conferring full protection against lethal challenge with SARS-CoV-2 in a transgenic mouse model.The neutralizing antibodies elicited by d16 effectively cross-reacted with several SARS-CoV-2 variants.Secretory immunoglobulin A was detected in the blood and nasal wash of vaccinated mice.Our work provides proof-of-principle evidence for harnessing NSP16-deficient SARS-CoV-2 for the development of live attenuated vaccines and paves the way for further preclinical studies of d16 as a prototypic vaccine strain,to which new features might be introduced to improve safety,transmissibility,immunogenicity and efficacy.
基金supported by the Consultancy Service for Enhancing Laboratory Surveillance of Emerging Infectious Disease for Department of Health of the Hong Kong Special Administrative Region of China,the Research Grant Council Grant,the University Development Fund,the Outstanding Young Researcher Award,and the Committee for Research and Conference Grant and The University of Hong Kong,China
文摘A straight, non-sporulating, Gram-variable bacillus (HKU24T) was recovered from the blood culture of a patient with metastatic breast carcinoma. After repeated subculturing in BACTEC Plus Anaerobic/F blood culture broth, HKU24T grew on brucella agar as non-hemolytic, pinpoint colonies after 96 h of incubation at 37 °C in an anaerobic environment and aerobic environment with 5% CO2. Growth was enhanced with a streak of Staphylococcus aureus. HKU24T was non-motile and catalase-negative, but positive for alkaline phosphatase, β-glucosidase, and α-glucosidase. It hydrolyzed phenylphosphonate and reduced resazurin. 16S rRNA, groEL, gyrB, recA, and rpoB sequencing showed that HKU24T occupies a distinct phylogenetic position among the Leptotrichia species, being most closely related to Leptotrichia trevisanii. Using HKU24T groEL, gyrB, recA, and rpoB gene-specific primers, fragments of these genes were amplified from one of 20 oral specimens. Based on phenotypic and genotypic characteristics, we propose a new species, Leptotrichia hongkongensis sp. nov., to describe this bacterium.
基金supported by the National Key Research and Development Program of China(2017YFC1200202)the National Natural Science Foundation of China(81290342,81461168030)+3 种基金the Major Special Projects for Infectious Disease Research of China(2016ZX10004222-003)China National Grand S&T Special Project(2014ZX10004-001-006)supported by CAS-TWAS President’s Fellowship of the University of Chinese Academy of Sciences(UCAS)and The World Academy of Sciences(TWAS)a leading principle investigator of the NSFC Innovative Research Group(81621091)
文摘Bats are connected with the increasing numbers of emerging and re-emerging viruses that may break the species barrier and spread into the human population. Coronaviruses are one of the most common viruses discovered in bats, which were considered as the natural source of recent human-susceptible coronaviruses, i.e. SARS-COV and MERS-CoV. Our previous study reported the discovery of a bat-derived putative cross-family recombinant coronavirus with a reovirus gene p10, named as Ro-BatCoV GCCDC1. In this report, through a two-year follow-up of a special bat population in one specific cave of south China, we illustrate that Ro-BatCoV GCCDC1 persistently circulates among bats. Notably, through the longitudinal observation, we identified the dynamic evolution of Ro-BatCoV GCCDC1 in bats represented by continuously recombination events. Our study provides the first glimpse of the virus evolution in one longitudinally observed bat population cohort and underlines the surveillance and pre-warning of potential interspecies transmittable viruses in bats.
文摘Avian influenza viruses have been first known to jump the interspecies barrier and cause human infections since 1959[1].However,avian influenza was not considered a serious threat to public health until 1997,when the influenza A(H5N1)virus caused an outbreak in Hong Kong with 18 cases including six deaths[2].The A(H5N1)epidemic has never stopped despite poultry immu-
文摘The pandemic H1N12009 influenza virus has caused thefirst influenza pandemic of the 21st century,leading to disproportionate fatalities in the low-risk population despite the generally mild nature of the illness.Advances in science and technology have allowed very detailed study on the pathogenesis of this novel virus,and many have already been published in less than a year after the start of the pandemic.Information generated from cell lines,animal models,and clinical data analysis has provided us with greater understanding of the behavior of this virus and the associated host response.The new knowledge will allow us to formulate scientifically sound and evidence-based management plans.
基金partly supported by funding from Health@InnoHK,Innovation and Technology Commission,the Government of the Hong Kong Special Administrative RegionTheme-Based Research Scheme of the Research Grants Council(T11-709/21-N)+7 种基金the National Program on Key Research Project of China(2020YFA0707500 and 2020YFA0707504)Guangdong Natural Science Foundation(2022A1515010099)the University of Hong Kong Outstanding Young Researcher Awardthe University of Hong Kong Li Ka Shing Faculty of Medicine Research Output Prizethe High Level-Hospital Program,Health Commission of Guangdong Province,Chinathe research project of Hainan Academician Innovation Platform(YSPTZX202004)Emergency Key Program of Guangzhou Laboratory(EKPG22-01)the Swiss National Science Foundation,the National Research Programme Covid-19(No.4078P0_198290/1)。
文摘The emergence of SARS-CoV-2 variants of concern and repeated outbreaks of coronavirus epidemics in the past two decades emphasize the need for next-generation pan-coronaviral therapeutics.Drugging the multi-functional papain-like protease(PLpro)domain of the viral nsp3 holds promise.However,none of the known coronavirus PLpro inhibitors has been shown to be in vivo active.Herein,we screened a structurally diverse library of 50,080 compounds for potential coronavirus PLpro inhibitors and identified a noncovalent lead inhibitor F0213 that has broad-spectrum anti-coronaviral activity,including against the Sarbecoviruses(SARSCoV-1 and SARS-CoV-2),Merbecovirus(MERS-CoV),as well as the Alphacoronavirus(hCoV-229E and hCoVOC43).Importantly,F0213 confers protection in both SARS-CoV-2-infected hamsters and MERS-CoV-infected human DPP4-knockin mice.F0213 possesses a dual therapeutic functionality that suppresses coronavirus replication via blocking viral polyprotein cleavage,as well as promoting antiviral immunity by antagonizing the PLpro deubiquitinase activity.Despite the significant difference of substrate recognition,mode of inhibition studies suggest that F0213 is a competitive inhibitor against SARS2-PLpro via binding with the 157K amino acid residue,whereas an allosteric inhibitor of MERSPLpro interacting with its 271E position.Our proof-ofconcept findings demonstrated that PLpro is a valid target for the development of broad-spectrum anticoronavirus agents.The orally administered F0213 may serve as a promising lead compound for combating the ongoing COVID-19 pandemic and future coronavirus outbreaks.
基金This study was supported by fundings from Health@InnoHK,Innovation and Technology Commission,the Government of the Hong Kong Special Administrative Regionthe Health and Medical Research Fund,the Government of the Hong Kong Special Administrative Region(COVID1903010 Project 13)+3 种基金the National Program on Key Research Project of China(2020YFA0707500)Theme-Based Research Scheme of the Research Grants Council(11-709/21-N),Emergency Key Program of Guangzhou Laboratory(EKPG22-01)the National Key Research and Development Programme on Public Security Risk Prevention and Control Emergency Project,the Health and Medical Research Fund(22210792)donations of Michael Seak-Kan Tong,Richard Yu and Carol Yu,Lee Wan Keung Charity Foundation Limited,May Tam Mak Mei Yin,Respiratory Viral Research Foundation Limited,the Shaw Foundation Hong Kong,Hui Ming,Hui Hoy and Chow Sin Lan Charity Fund Limited,Chan Yin Chuen Memorial Charitable Foundation,The Chen Wai Wai Vivien Foundation Limited,and Marina Man-Wai Lee.We thanks Dr Jenny K.W.Lam for providing LAH4 for the experiments.
文摘Defective interfering genes(DIGs)are short viral genomes and interfere with wild-type viral replication.Here,we demonstrate that the new designed SARS-CoV-2 DIG(CD3600)can significantly inhibit the replication of SARS-CoV-2 including Alpha,Delta,Kappa and Omicron variants in human HK-2 cells and influenza DIG(PAD4)can significantly inhibit influenza virus replication in human A549 cells.One dose of influenza DIGs prophylactically protects 90%mice from lethal challenge of A(H1N1)pdm09 virus and CD3600 inhibits SARS-CoV-2 replication in hamster lungs when DIGs are administrated to lungs one day before viral challenge.To further investigate the gene delivery vector in the respiratory tract,a peptidic TAT2-P1&LAH4,which can package genes to form small spherical nanoparticles with high endosomal escape ability,is demonstrated to dramatically increase gene expression in the lung airway.TAT2-P1&LAH4,with the dual-functional TAT2-P1(gene-delivery and antiviral),can deliver CD3600 to significantly inhibit the replication of Delta and Omicron SARS-CoV-2 in hamster lungs.This peptide-based nanoparticle system can effectively transfect genes in lungs and deliver DIGs to inhibit SARS-CoV-2 variants and influenza virus in vivo,which provides the new insight into the drug delivery system for gene therapy against respiratory viruses.