用NDV La Sota株、IBV M41株、EDS76 V HSH23株、AEV Van Roekel株为种毒,分别接种鸡胚或鸭胚,制备各种病毒抗原液,经福尔马林灭活后,采用FILTRON盒式超滤系统对制苗病毒抗原液进行浓缩,然后按一定比例混合,以司本80及吐温80为乳化剂,1...用NDV La Sota株、IBV M41株、EDS76 V HSH23株、AEV Van Roekel株为种毒,分别接种鸡胚或鸭胚,制备各种病毒抗原液,经福尔马林灭活后,采用FILTRON盒式超滤系统对制苗病毒抗原液进行浓缩,然后按一定比例混合,以司本80及吐温80为乳化剂,10号白油为佐剂,制成ND-IB-EDS76-AE四联灭活疫苗。对四联苗各项技术指标进行了测定,证明本疫苗安全、无任何副作用;免疫10-14 d后产生免疫力;ND部分效检,攻毒100％保护,每羽份含ND效价达50-123 PD50;IB部分效检,免疫21-28 d后攻毒保护率达80％-100％,IB HI效价≥1:64;EDS76效检,免疫后21 d HI效价≥1:128;AE效检,保护率达80％-100％。展开更多
Newcastle disease virus(NDV)and H9N2 subtype Avian influenza virus(AIV)are two notorious avian respiratory pathogens that cause great losses in the poultry industry.Current inactivated commercial vaccines against NDV ...Newcastle disease virus(NDV)and H9N2 subtype Avian influenza virus(AIV)are two notorious avian respiratory pathogens that cause great losses in the poultry industry.Current inactivated commercial vaccines against NDV and AIV have the disadvantages of inadequate mucosal responses,while an attenuated live vaccine bears the risk of mutation.Dendritic cell(DC)targeting strategies are attractive for their potent mucosal and adaptive immune-stimulating ability against respiratory pathogens.In this study,DC-binding peptide(DCpep)-decorated chimeric virus-like particles(cVLPs),containing NDV haemagglutinin–neuraminidase(HN)and AIV haemagglutinin(HA),were developed as a DC-targeting mucosal vaccine candidate.DCpep-decorated cVLPs activated DCs in vitro,and induced potent immune stimulation in chickens,with enhanced secretory immunoglobulin A(sIgA)secretion and splenic T cell differentiation.40μg cVLPs can provide full protection against the challenge with homologous,heterologous NDV strains,and AIV H9N2.In addition,DCpep-decorated cVLPs could induce a better immune response when administered intranasally than intramuscularly,as indicated by robust s IgA secretion and a reduced virus shedding period.Taken together,this chimericVLPs are a promising vaccine candidate to control NDV and AIV H9N2 and a useful platform bearing multivalent antigens.展开更多
As a potential vectored vaccine,Newcastle disease virus(NDV)has been subject to various studies for vaccine development,while relatively little research has outlined the immunomodulatory effect of the virus in antigen...As a potential vectored vaccine,Newcastle disease virus(NDV)has been subject to various studies for vaccine development,while relatively little research has outlined the immunomodulatory effect of the virus in antigen presentation.To elucidate the key inhibitory factor in regulating the interaction of infected dendritic cells(DCs)and T cells,DCs were pretreated with the NDV vaccine strain LaSota as an inhibitor and stimulated with lipopolysaccharide(LPS)for further detection by enzyme-linked immunosorbent assay(ELISA),flow cytometry,immunoblotting,and quantitative real-time polymerase chain reaction(qRT-PCR).The results revealed that NDV infection resulted in the inhibition of interleukin(IL)-12p40 in DCs through a p38 mitogen-activated protein kinase(MAPK)-dependent manner,thus inhibiting the synthesis of IL-12p70,leading to the reduction in T cell proliferation and the secretion of interferon-(IFN-),tumor necrosis factor-α(TNF-α),and IL-6 induced by DCs.Consequently,downregulated cytokines accelerated the infection and viral transmission from DCs to T cells.Furthermore,several other strains of NDV also exhibited inhibitory activity.The current study reveals that NDV can modulate the intensity of the innate-adaptive immune cell crosstalk critically toward viral invasion improvement,highlighting a novel mechanism of virus-induced immunosuppression and providing new perspectives on the improvement of NDV-vectored vaccine.展开更多
Breast cancer,an unceasingly occurring neoplasm,is one of the major determinants of mortality in women.Several ineffective attempts have been pursued using with conventional therapies against breast cancer.Resistance ...Breast cancer,an unceasingly occurring neoplasm,is one of the major determinants of mortality in women.Several ineffective attempts have been pursued using with conventional therapies against breast cancer.Resistance to existing therapies and their respective debilitating adverse effects have led research toward a new era of cancer treatment using viruses.Virotherapy constitutes a developing treatment modality with multiple mechanisms of therapeutic activity in which the viruses can be directly oncolyticand can express transgenes or induce host immune response against tumor cells.Several different DNA-and RNA-containing viruses have been considered for virotherapy of breast cancer including adenovirus,herpes virus,vaccinia,reovirus,Newcastle Disease virus,measles virus and vesicular stomatitis virus.This review aims to summarize the viro-therapeutical agents against breast malignancies.Key Scientific Concepts of Review:In this review paper,we proposed a new strategy to virus's combinatorial treatments using several kinds of transgenes and drugs.These recombinant viruses have provided evidence of treatment efficacy against human breast cancer.展开更多
Background and Aims:Hepatocellular carcinoma(HCC)is listed as one of the most common causes of cancer-related death.Oncolytic therapy has become a promising treatment because of novel immunotherapies and gene editing ...Background and Aims:Hepatocellular carcinoma(HCC)is listed as one of the most common causes of cancer-related death.Oncolytic therapy has become a promising treatment because of novel immunotherapies and gene editing technology,but biosafety concerns remain the biggest limitation for clinical application.We studied the the antitumor activity and biosafety of the wild-type Newcastle disease virus HK84 strain(NDV/HK84)and 10 other NDV strains.Meth-ods:Cell proliferation and apoptosis were determined by cell counting Kit-8 and fluorescein isothiocyanate Annexin V apoptosis assays.Colony formation,wound healing,and a xenograft mouse model were used to evaluate in vivo and in vitro oncolytic effectiveness.The safety of NDV/HK84 was tested in nude mice by an in vivo luciferase imaging system.The replication kinetics of NDV/HK84 in normal tis-sues and tumors were evaluated by infectious-dose assays in eggs.RNA sequencing analysis was performed to explore NDV/HK84 activity and was validated by quantitative real-time PCR.Results:The cell counting Kit-8 assays of vi-ability found that the oncolytic activity of the NDV strains differed with the multiplicity of infection(MOI).At an MOI of 20,the oncolytic activity of all NDV strains except the DK/JX/21358/08 strain was>80%.The oncolytic activities of the NDV/HK84 and DK/JX/8224/04 strains were>80%at both MOI=20 and MOI=2.Only NDV/HK84 had>80%oncolytic activities at both MOI=20 and MOI=2.We chose NDV/HK84 as the candidate virus to test the oncolytic effect of NDV in HCC in the in vitro and in vivo experiments.NDV/HK84 killed human SK-HEP-1 HCC cells without affecting healthy cells.Conclusions:Intratumor infection with NDV/HK84 strains compared with vehicle controls or positive controls indicated that NDV/HK84 strain specifically inhib-ited HCC without affecting healthy mice.High-throughput RNA sequencing showed that the oncolytic activity of NDV/HK84 was dependent on the activation of type I interferon signaling.展开更多
新城疫和禽流感严重危害世界养禽业,对其防控在我国主要依赖疫苗接种。课题组前期获得了一株可表达H9亚型禽流感病毒HA蛋白的重组耐热新城疫病毒rTS-HA株。进一步对其鸡胚传代特性进行研究,旨在分析重组新城疫病毒在传代过程中的热稳定...新城疫和禽流感严重危害世界养禽业,对其防控在我国主要依赖疫苗接种。课题组前期获得了一株可表达H9亚型禽流感病毒HA蛋白的重组耐热新城疫病毒rTS-HA株。进一步对其鸡胚传代特性进行研究,旨在分析重组新城疫病毒在传代过程中的热稳定性和外源基因表达稳定性。将其在SPF鸡胚传至18代,每隔4代耐热选育1次,取中间代次毒株,测定其增殖效价、鸡胚最小致死剂量的平均死亡时间(mean death time,MDT),同时设置未耐热选育的对照。结果表明,通过耐热选育将rTS-HA株传至第18代后,其增殖滴度未发现显著变化,MDT均大于120 h,表明毒力无返强现象,但其耐热性有显著提升,而未进行耐热选育的鸡胚传代株的耐热特性显著下降。动物试验结果表明,耐热选育后毒株的免疫效果较初代毒株也有明显的提升。以上结果表明重组新城疫病毒rTS-HA株需要进行耐热选育传代,才能保持其耐热特性,同时其免疫原性也有一定的提升,可作为新城疫和禽流感二联耐热基因工程活疫苗的候选毒株。展开更多
The influence of antibody immune selective pressure on Newcastle disease virus (NDV) HN and F gene mutations was studied in cell cultures.NDV field strain TZ060107 was inoculated into chicken embryo fibroblast cells a...The influence of antibody immune selective pressure on Newcastle disease virus (NDV) HN and F gene mutations was studied in cell cultures.NDV field strain TZ060107 was inoculated into chicken embryo fibroblast cells and continuously passaged with (group A) or without (group B) anti-NDV monospecific serum.Each group contained three independent passage series.HN and F genes were amplified and sequenced for the 10th,20th,30th,40th and 50th generations of each serial passage,and compared with the original strain.The results demonstrated that increased HN gene mutations were observed in group A with the antibody than in group B without the antibody.The nonsynonymous (NS) to synonymous (S) mutations ratio was 6 for group A,significantly higher than 3.4 in group B.In group A with the antibody,there were five stable NS mutations in HN gene,three of which (related to aa#353,521 and 568) were related to known epitopes.There were two stable NS mutations in F gene in group A,but no stable NS mutations in group B.The NS/S ratios of F gene were less than 2.5 for both groups A and B.Our results suggested that the antibody strongly influenced HN gene mutations,while the F gene was less influenced by the same antibody.展开更多
Dear Editor, Newcastle disease virus （NDV）, also known as avian paramyxovirus serotype 1 （APMV-1）, is a member of the genus Avulavirus within the family Paramyxoviridae, or- der Mononegavirales （Miller et al., 2...Dear Editor, Newcastle disease virus （NDV）, also known as avian paramyxovirus serotype 1 （APMV-1）, is a member of the genus Avulavirus within the family Paramyxoviridae, or- der Mononegavirales （Miller et al., 2010）. Although all isolated NDV strains belong to a single serotype, epi- demiological studies have revealed that the genotype VII strain is currently the most prevalent circulating geno- type worldwide and is associated with many of the most recent outbreaks in China since 1997 （Liu et al., 2007）.展开更多
文摘用NDV La Sota株、IBV M41株、EDS76 V HSH23株、AEV Van Roekel株为种毒,分别接种鸡胚或鸭胚,制备各种病毒抗原液,经福尔马林灭活后,采用FILTRON盒式超滤系统对制苗病毒抗原液进行浓缩,然后按一定比例混合,以司本80及吐温80为乳化剂,10号白油为佐剂,制成ND-IB-EDS76-AE四联灭活疫苗。对四联苗各项技术指标进行了测定,证明本疫苗安全、无任何副作用;免疫10-14 d后产生免疫力;ND部分效检,攻毒100％保护,每羽份含ND效价达50-123 PD50;IB部分效检,免疫21-28 d后攻毒保护率达80％-100％,IB HI效价≥1:64;EDS76效检,免疫后21 d HI效价≥1:128;AE效检,保护率达80％-100％。
基金supported by grants from the National Key Research and Development Program of China(Grant No.2018YFD0500100)the National Natural Science Foundation of China(Grant Nos.31772735,31472195)+2 种基金the Jiangsu Provincial Natural Science Foundation of China(Grant No.BK20180299)Jiangsu Agriculture Science and Technology Innovation Fund CX(19)3019the Key Technology Research and Development Project of Jilin Province(Grant No.20180201021NY)。
文摘Newcastle disease virus(NDV)and H9N2 subtype Avian influenza virus(AIV)are two notorious avian respiratory pathogens that cause great losses in the poultry industry.Current inactivated commercial vaccines against NDV and AIV have the disadvantages of inadequate mucosal responses,while an attenuated live vaccine bears the risk of mutation.Dendritic cell(DC)targeting strategies are attractive for their potent mucosal and adaptive immune-stimulating ability against respiratory pathogens.In this study,DC-binding peptide(DCpep)-decorated chimeric virus-like particles(cVLPs),containing NDV haemagglutinin–neuraminidase(HN)and AIV haemagglutinin(HA),were developed as a DC-targeting mucosal vaccine candidate.DCpep-decorated cVLPs activated DCs in vitro,and induced potent immune stimulation in chickens,with enhanced secretory immunoglobulin A(sIgA)secretion and splenic T cell differentiation.40μg cVLPs can provide full protection against the challenge with homologous,heterologous NDV strains,and AIV H9N2.In addition,DCpep-decorated cVLPs could induce a better immune response when administered intranasally than intramuscularly,as indicated by robust s IgA secretion and a reduced virus shedding period.Taken together,this chimericVLPs are a promising vaccine candidate to control NDV and AIV H9N2 and a useful platform bearing multivalent antigens.
文摘As a potential vectored vaccine,Newcastle disease virus(NDV)has been subject to various studies for vaccine development,while relatively little research has outlined the immunomodulatory effect of the virus in antigen presentation.To elucidate the key inhibitory factor in regulating the interaction of infected dendritic cells(DCs)and T cells,DCs were pretreated with the NDV vaccine strain LaSota as an inhibitor and stimulated with lipopolysaccharide(LPS)for further detection by enzyme-linked immunosorbent assay(ELISA),flow cytometry,immunoblotting,and quantitative real-time polymerase chain reaction(qRT-PCR).The results revealed that NDV infection resulted in the inhibition of interleukin(IL)-12p40 in DCs through a p38 mitogen-activated protein kinase(MAPK)-dependent manner,thus inhibiting the synthesis of IL-12p70,leading to the reduction in T cell proliferation and the secretion of interferon-(IFN-),tumor necrosis factor-α(TNF-α),and IL-6 induced by DCs.Consequently,downregulated cytokines accelerated the infection and viral transmission from DCs to T cells.Furthermore,several other strains of NDV also exhibited inhibitory activity.The current study reveals that NDV can modulate the intensity of the innate-adaptive immune cell crosstalk critically toward viral invasion improvement,highlighting a novel mechanism of virus-induced immunosuppression and providing new perspectives on the improvement of NDV-vectored vaccine.
文摘Breast cancer,an unceasingly occurring neoplasm,is one of the major determinants of mortality in women.Several ineffective attempts have been pursued using with conventional therapies against breast cancer.Resistance to existing therapies and their respective debilitating adverse effects have led research toward a new era of cancer treatment using viruses.Virotherapy constitutes a developing treatment modality with multiple mechanisms of therapeutic activity in which the viruses can be directly oncolyticand can express transgenes or induce host immune response against tumor cells.Several different DNA-and RNA-containing viruses have been considered for virotherapy of breast cancer including adenovirus,herpes virus,vaccinia,reovirus,Newcastle Disease virus,measles virus and vesicular stomatitis virus.This review aims to summarize the viro-therapeutical agents against breast malignancies.Key Scientific Concepts of Review:In this review paper,we proposed a new strategy to virus's combinatorial treatments using several kinds of transgenes and drugs.These recombinant viruses have provided evidence of treatment efficacy against human breast cancer.
基金supported by research grants from the Guangdong Science and Technology Innovation Strategy Special Found(2019B121205009)the Guangdong Science and Technology Special Found(190830095586328 and 200109155890863)and the Li Ka Shing Foundation.
文摘Background and Aims:Hepatocellular carcinoma(HCC)is listed as one of the most common causes of cancer-related death.Oncolytic therapy has become a promising treatment because of novel immunotherapies and gene editing technology,but biosafety concerns remain the biggest limitation for clinical application.We studied the the antitumor activity and biosafety of the wild-type Newcastle disease virus HK84 strain(NDV/HK84)and 10 other NDV strains.Meth-ods:Cell proliferation and apoptosis were determined by cell counting Kit-8 and fluorescein isothiocyanate Annexin V apoptosis assays.Colony formation,wound healing,and a xenograft mouse model were used to evaluate in vivo and in vitro oncolytic effectiveness.The safety of NDV/HK84 was tested in nude mice by an in vivo luciferase imaging system.The replication kinetics of NDV/HK84 in normal tis-sues and tumors were evaluated by infectious-dose assays in eggs.RNA sequencing analysis was performed to explore NDV/HK84 activity and was validated by quantitative real-time PCR.Results:The cell counting Kit-8 assays of vi-ability found that the oncolytic activity of the NDV strains differed with the multiplicity of infection(MOI).At an MOI of 20,the oncolytic activity of all NDV strains except the DK/JX/21358/08 strain was>80%.The oncolytic activities of the NDV/HK84 and DK/JX/8224/04 strains were>80%at both MOI=20 and MOI=2.Only NDV/HK84 had>80%oncolytic activities at both MOI=20 and MOI=2.We chose NDV/HK84 as the candidate virus to test the oncolytic effect of NDV in HCC in the in vitro and in vivo experiments.NDV/HK84 killed human SK-HEP-1 HCC cells without affecting healthy cells.Conclusions:Intratumor infection with NDV/HK84 strains compared with vehicle controls or positive controls indicated that NDV/HK84 strain specifically inhib-ited HCC without affecting healthy mice.High-throughput RNA sequencing showed that the oncolytic activity of NDV/HK84 was dependent on the activation of type I interferon signaling.
文摘新城疫和禽流感严重危害世界养禽业,对其防控在我国主要依赖疫苗接种。课题组前期获得了一株可表达H9亚型禽流感病毒HA蛋白的重组耐热新城疫病毒rTS-HA株。进一步对其鸡胚传代特性进行研究,旨在分析重组新城疫病毒在传代过程中的热稳定性和外源基因表达稳定性。将其在SPF鸡胚传至18代,每隔4代耐热选育1次,取中间代次毒株,测定其增殖效价、鸡胚最小致死剂量的平均死亡时间(mean death time,MDT),同时设置未耐热选育的对照。结果表明,通过耐热选育将rTS-HA株传至第18代后,其增殖滴度未发现显著变化,MDT均大于120 h,表明毒力无返强现象,但其耐热性有显著提升,而未进行耐热选育的鸡胚传代株的耐热特性显著下降。动物试验结果表明,耐热选育后毒株的免疫效果较初代毒株也有明显的提升。以上结果表明重组新城疫病毒rTS-HA株需要进行耐热选育传代,才能保持其耐热特性,同时其免疫原性也有一定的提升,可作为新城疫和禽流感二联耐热基因工程活疫苗的候选毒株。
基金supported by the Shandong Province Application Technology Innovation Project on Agriculture during 2007 and 2008
文摘The influence of antibody immune selective pressure on Newcastle disease virus (NDV) HN and F gene mutations was studied in cell cultures.NDV field strain TZ060107 was inoculated into chicken embryo fibroblast cells and continuously passaged with (group A) or without (group B) anti-NDV monospecific serum.Each group contained three independent passage series.HN and F genes were amplified and sequenced for the 10th,20th,30th,40th and 50th generations of each serial passage,and compared with the original strain.The results demonstrated that increased HN gene mutations were observed in group A with the antibody than in group B without the antibody.The nonsynonymous (NS) to synonymous (S) mutations ratio was 6 for group A,significantly higher than 3.4 in group B.In group A with the antibody,there were five stable NS mutations in HN gene,three of which (related to aa#353,521 and 568) were related to known epitopes.There were two stable NS mutations in F gene in group A,but no stable NS mutations in group B.The NS/S ratios of F gene were less than 2.5 for both groups A and B.Our results suggested that the antibody strongly influenced HN gene mutations,while the F gene was less influenced by the same antibody.
基金supported by the Innovation Foundation Project of China Animal Health and Epidemiology Center,Ministry of Agriculture,P.R.China(project no:CAHEC-2015-Y105)supported by the National Natural Scientific Fund(31272561)
文摘Dear Editor, Newcastle disease virus （NDV）, also known as avian paramyxovirus serotype 1 （APMV-1）, is a member of the genus Avulavirus within the family Paramyxoviridae, or- der Mononegavirales （Miller et al., 2010）. Although all isolated NDV strains belong to a single serotype, epi- demiological studies have revealed that the genotype VII strain is currently the most prevalent circulating geno- type worldwide and is associated with many of the most recent outbreaks in China since 1997 （Liu et al., 2007）.
