Viral co-infection has been found in animals;however,the mechanisms of co-infection are unclear.The abundance and diversity of viruses in water make fish highly susceptible to co-infection.Here,we reported a coinfecti...Viral co-infection has been found in animals;however,the mechanisms of co-infection are unclear.The abundance and diversity of viruses in water make fish highly susceptible to co-infection.Here,we reported a coinfection in fish,which resulted in reduced host lethality and illustrated the intracellular molecular mechanism of viral co-infection.The spring viremia of carp virus(SVCV)is a highly lethal virus that infects Cyprinidae,such as zebrafish.The mortality of SVCV infection was significantly reduced when co-infected with the grass carp reovirus(GCRV).The severity of tissue damage and viral proliferation of SVCV was also reduced in co-infection with GCRV.The transcriptome bioinformatics analysis demonstrated that the effect on the host transcripts in response to SVCV infection was significantly reduced in co-infection.After excluding the extracellular interactions of these two viruses,the intracellular mechanisms were studied.We found that the GCRV NS38 remarkably decreased SVCV infection and viral proliferation.The interaction between GCRV NS38 and SVCV nucleoprotein(N)and phosphoprotein(P)proteins was identified,and NS38 downregulated both N and P proteins.Further analysis demonstrated that the N protein was degraded by NS38 indispensable of the autophagy receptor,sequestosome 1(p62).Meanwhile,K63-linked ubiquitination of the P protein was reduced by NS38,leading to ubiquitinated degradation of the P protein.These results reveal that the intracellular viral protein interactions are a crucial mechanism of co-infection and influence the host pathology and expand our understanding in intracellular viral interactions co-infection.展开更多
Since the anti-inflammatory effect of hydrogen has been widely known,it was supposed that hydrogen could suppress tissue damage by inhibiting virus-related inflammatory reactions.However,hydrogen is slightly soluble i...Since the anti-inflammatory effect of hydrogen has been widely known,it was supposed that hydrogen could suppress tissue damage by inhibiting virus-related inflammatory reactions.However,hydrogen is slightly soluble in water,which leads to poor effect of oral hydrogen-rich water therapy.In this study,the nano-bubble hydrogen water(nano-HW)(about 0.7 ppm)was prepared and its therapeutic effect against viral infection was investigated by utilizing spring viraemia of carp virus(SVCV)-infected zebrafish as model.Three-month-old zebrafish were divided into nano-HW treatment-treated group and aquaculture water treated group(control group).The results revealed that the cumulative mortality rate of SVCV-infected zebrafish was reduced by 40%after treatment with nano-bubble hydrogen water,and q RT-PCR results showed that SVCV replication was significantly inhibited.Histopathological examination staining showed that SVCV infection caused tissue damage was greatly alleviated after treatment with nano-bubble hydrogen water.Futhermore,SVCV infection caused reactive oxygen species(ROS)accumulation was significantly reduced upon nano-HW treatment.The level of proinflammatory cytokines IL-1β,IL-8,and TNF-αwas remarkably reduced in the nano-HW-treated group in vivo and in vitro.Taken together,our data demonstrated for the first time that nano-HW could inhibit the inflammatory response caused by viral infection in zebrafish,which suggests that nano-HW can be applied to antiviral research,and provides a novel therapeutic strategy for virus-caused inflammation related disease.展开更多
[Objective]The paper was to develop a rapid method for the detection of spring Viremia of carp virus(SVCV).[Method]The specific primers were designed by targeting the G gene of SVCV.The recombinase polymerase amplific...[Objective]The paper was to develop a rapid method for the detection of spring Viremia of carp virus(SVCV).[Method]The specific primers were designed by targeting the G gene of SVCV.The recombinase polymerase amplification(RPA)assay for detecting SVCV was estab-lished by optimizing the reaction conditions.The optimal amplification temperature of RPA assay was 30℃,and the test could be finished within 20 min.[Result]The method was specific with no cross-reaction with other common fish infectious viruses.Sensitivity test showed that the lowest detection limit of the method was 89.2 copies/μL,higher than that of traditional RT-PCR.Moreover,a total of 80 clinical samples were detected by RPA and RT-PCR,respectively.The weak positive samples tested by RT-PCR could be detectable with RPA,indicating that RPA assay could be used in clinical detection.[Conclusion]The method established is rapid,simple,specific and sensitive for testing SVCV,and it will be widely used in grassroots laboratory and on-site inspection.展开更多
为了揭示鲤春病毒血症病毒（spring viremia of carp virus,SVCV）糖蛋白编码基因的主要免疫原性决定区域,本研究对SVCV的糖蛋白基因进行截短表达,并用纯化的重组蛋白作为免疫原制备了兔抗血清以分析其免疫原性。通过SOSUI以及DNAStar ...为了揭示鲤春病毒血症病毒（spring viremia of carp virus,SVCV）糖蛋白编码基因的主要免疫原性决定区域,本研究对SVCV的糖蛋白基因进行截短表达,并用纯化的重组蛋白作为免疫原制备了兔抗血清以分析其免疫原性。通过SOSUI以及DNAStar 6.0软件对SVCV糖蛋白基因的跨膜区、亲水性以及抗原表位进行分析后,采用RT-PCR对该基因主要抗原决定区域编码片段进行扩增,并构建重组表达质粒p GEX-Gtr,对其进行诱导表达后获得截短的SVCV糖蛋白的重组蛋白。将表达的重组蛋白进行纯化复性后,作为免疫原制备兔抗血清,采用ELISA法检测其效价,采用免疫印迹以及间接免疫荧光技术检测该重组蛋白的免疫原性。研究结果显示,截短表达的糖蛋白编码基因长1317 bp,编码439个氨基酸（29~467）,推测分子量为49.6 k D。利用该重组蛋白制备的兔抗血清,其与重组蛋白的反应效价为1∶64000,。免疫印迹及间接免疫荧光结果显示,该兔抗血清与SVCV-HN株能发生特异性反应,表明该重组蛋白与天然的病毒表面糖蛋白的免疫原性无差异。上述研究结果表明,截短表达的重组蛋白具有很好的免疫原性,可应用于免疫诊断技术研发与基因工程疫苗的研制。展开更多
Tripartite motif(TRIM)proteins were shown to play an important role in innate antiviral immunity.FinTRIM(ftr)is a new subset of TRIM genes that do not possess obvious orthologs in higher vertebrates.However,little is ...Tripartite motif(TRIM)proteins were shown to play an important role in innate antiviral immunity.FinTRIM(ftr)is a new subset of TRIM genes that do not possess obvious orthologs in higher vertebrates.However,little is known about its function.In this study,we used bioinformatic analysis to examine the phylogenetic relationships and conserved domains of zebrafish(Danio rerio)fir01,fir42,and fir58.as well as qualitative real-time PCR to examine their expression patterns in zebrafish embryonic fibroblast(ZF4)cells and zebrafish tissues.Sequence analysis showed that the three finTRIMs are highly conserved,and all contain a RING domain.B-box domain,and SPRY-PRY domain.In addition,fr42 and fir58 had one coiled-coil domain(CCD),whereas ftrol had two CCDs.Tissue expression analysis revealed that the mRNA level of ftr01 was the highest in the liver,whereas those of ftr42 and ftr58 were the highest in the gill;the expression of thesefinTRIMs was clearly upregulated not in the eyes,but in the liver,spleen,kidney,gill,and brain of zebrafish following spring viremia of carp virus(SVCV)infection.Similarly,the expression of these three finTRIM genes also increased in ZF4 cells after SVCV infection.Our study revealed that fir01,fr42,and fir58 may play an important role in antiviralimmune responses,and these findings validate the need for more in-depth research on the finTRIM family in the future.展开更多
基金supported by the Strategic Priority Research Program of the Chinese Academy of Sciences(XDA24030203)the National Key Research and Development Program of China(2018YFD0900504,2018YFD0900204,and 2021YFD1200804)+3 种基金the National Natural Science Foundation of China(32073009,31873036,32173023,and 32002431)the Youth Innovation Promotion Association,the Key Program of Frontier Sciences of the Chinese Academy of Sciences(QYZDY-SSW-SMC025)the China Agriculture Research System of MOF and MARA(CARS-45-07)the Autonomous Project of the State Key Laboratory of Freshwater Ecology and Biotechnology.
文摘Viral co-infection has been found in animals;however,the mechanisms of co-infection are unclear.The abundance and diversity of viruses in water make fish highly susceptible to co-infection.Here,we reported a coinfection in fish,which resulted in reduced host lethality and illustrated the intracellular molecular mechanism of viral co-infection.The spring viremia of carp virus(SVCV)is a highly lethal virus that infects Cyprinidae,such as zebrafish.The mortality of SVCV infection was significantly reduced when co-infected with the grass carp reovirus(GCRV).The severity of tissue damage and viral proliferation of SVCV was also reduced in co-infection with GCRV.The transcriptome bioinformatics analysis demonstrated that the effect on the host transcripts in response to SVCV infection was significantly reduced in co-infection.After excluding the extracellular interactions of these two viruses,the intracellular mechanisms were studied.We found that the GCRV NS38 remarkably decreased SVCV infection and viral proliferation.The interaction between GCRV NS38 and SVCV nucleoprotein(N)and phosphoprotein(P)proteins was identified,and NS38 downregulated both N and P proteins.Further analysis demonstrated that the N protein was degraded by NS38 indispensable of the autophagy receptor,sequestosome 1(p62).Meanwhile,K63-linked ubiquitination of the P protein was reduced by NS38,leading to ubiquitinated degradation of the P protein.These results reveal that the intracellular viral protein interactions are a crucial mechanism of co-infection and influence the host pathology and expand our understanding in intracellular viral interactions co-infection.
基金supported by Natural Science Foundation of China(31972834,32022082,31972721)。
文摘Since the anti-inflammatory effect of hydrogen has been widely known,it was supposed that hydrogen could suppress tissue damage by inhibiting virus-related inflammatory reactions.However,hydrogen is slightly soluble in water,which leads to poor effect of oral hydrogen-rich water therapy.In this study,the nano-bubble hydrogen water(nano-HW)(about 0.7 ppm)was prepared and its therapeutic effect against viral infection was investigated by utilizing spring viraemia of carp virus(SVCV)-infected zebrafish as model.Three-month-old zebrafish were divided into nano-HW treatment-treated group and aquaculture water treated group(control group).The results revealed that the cumulative mortality rate of SVCV-infected zebrafish was reduced by 40%after treatment with nano-bubble hydrogen water,and q RT-PCR results showed that SVCV replication was significantly inhibited.Histopathological examination staining showed that SVCV infection caused tissue damage was greatly alleviated after treatment with nano-bubble hydrogen water.Futhermore,SVCV infection caused reactive oxygen species(ROS)accumulation was significantly reduced upon nano-HW treatment.The level of proinflammatory cytokines IL-1β,IL-8,and TNF-αwas remarkably reduced in the nano-HW-treated group in vivo and in vitro.Taken together,our data demonstrated for the first time that nano-HW could inhibit the inflammatory response caused by viral infection in zebrafish,which suggests that nano-HW can be applied to antiviral research,and provides a novel therapeutic strategy for virus-caused inflammation related disease.
基金Supported by National Key Research and Development Program (2017YFF0211103)Scientific Research Project of General Administration of Quality Supervision,Inspection and Quarantine (2017IK232)
文摘[Objective]The paper was to develop a rapid method for the detection of spring Viremia of carp virus(SVCV).[Method]The specific primers were designed by targeting the G gene of SVCV.The recombinase polymerase amplification(RPA)assay for detecting SVCV was estab-lished by optimizing the reaction conditions.The optimal amplification temperature of RPA assay was 30℃,and the test could be finished within 20 min.[Result]The method was specific with no cross-reaction with other common fish infectious viruses.Sensitivity test showed that the lowest detection limit of the method was 89.2 copies/μL,higher than that of traditional RT-PCR.Moreover,a total of 80 clinical samples were detected by RPA and RT-PCR,respectively.The weak positive samples tested by RT-PCR could be detectable with RPA,indicating that RPA assay could be used in clinical detection.[Conclusion]The method established is rapid,simple,specific and sensitive for testing SVCV,and it will be widely used in grassroots laboratory and on-site inspection.
文摘为了揭示鲤春病毒血症病毒（spring viremia of carp virus,SVCV）糖蛋白编码基因的主要免疫原性决定区域,本研究对SVCV的糖蛋白基因进行截短表达,并用纯化的重组蛋白作为免疫原制备了兔抗血清以分析其免疫原性。通过SOSUI以及DNAStar 6.0软件对SVCV糖蛋白基因的跨膜区、亲水性以及抗原表位进行分析后,采用RT-PCR对该基因主要抗原决定区域编码片段进行扩增,并构建重组表达质粒p GEX-Gtr,对其进行诱导表达后获得截短的SVCV糖蛋白的重组蛋白。将表达的重组蛋白进行纯化复性后,作为免疫原制备兔抗血清,采用ELISA法检测其效价,采用免疫印迹以及间接免疫荧光技术检测该重组蛋白的免疫原性。研究结果显示,截短表达的糖蛋白编码基因长1317 bp,编码439个氨基酸（29~467）,推测分子量为49.6 k D。利用该重组蛋白制备的兔抗血清,其与重组蛋白的反应效价为1∶64000,。免疫印迹及间接免疫荧光结果显示,该兔抗血清与SVCV-HN株能发生特异性反应,表明该重组蛋白与天然的病毒表面糖蛋白的免疫原性无差异。上述研究结果表明,截短表达的重组蛋白具有很好的免疫原性,可应用于免疫诊断技术研发与基因工程疫苗的研制。
基金supported by the National Training Program of Innovation and Entrepreneurship for Undergraduates of Huazhong Agricultural University (2015308200403)the National Key Research and Development Program of China (2018YFD0900505)+1 种基金the Natural Science Foundation of China (31172433)the Fundamental Research Funds for the Central Universities (2662018YJ022)
文摘Tripartite motif(TRIM)proteins were shown to play an important role in innate antiviral immunity.FinTRIM(ftr)is a new subset of TRIM genes that do not possess obvious orthologs in higher vertebrates.However,little is known about its function.In this study,we used bioinformatic analysis to examine the phylogenetic relationships and conserved domains of zebrafish(Danio rerio)fir01,fir42,and fir58.as well as qualitative real-time PCR to examine their expression patterns in zebrafish embryonic fibroblast(ZF4)cells and zebrafish tissues.Sequence analysis showed that the three finTRIMs are highly conserved,and all contain a RING domain.B-box domain,and SPRY-PRY domain.In addition,fr42 and fir58 had one coiled-coil domain(CCD),whereas ftrol had two CCDs.Tissue expression analysis revealed that the mRNA level of ftr01 was the highest in the liver,whereas those of ftr42 and ftr58 were the highest in the gill;the expression of thesefinTRIMs was clearly upregulated not in the eyes,but in the liver,spleen,kidney,gill,and brain of zebrafish following spring viremia of carp virus(SVCV)infection.Similarly,the expression of these three finTRIM genes also increased in ZF4 cells after SVCV infection.Our study revealed that fir01,fr42,and fir58 may play an important role in antiviralimmune responses,and these findings validate the need for more in-depth research on the finTRIM family in the future.
