Nervous necrosis virus(NNV)can infect more than 120 fish species worldwide and has caused high mortality and sig-nificant economic losses to the aquaculture industry.Among different genotypes of NNV,the red-grouper ne...Nervous necrosis virus(NNV)can infect more than 120 fish species worldwide and has caused high mortality and sig-nificant economic losses to the aquaculture industry.Among different genotypes of NNV,the red-grouper nervous necrosis virus(RGNNV)is the most widely distributed one with the highest number of susceptible fish species.In this study,the capsid protein(Cp)gene of RGNNV was recombined and expressed in Escherichia coli strain BL21(DE3)and the recombinant Cp(rCp)was used as an immunogen to produce monoclonal antibodies(MAbs)through hybridoma cell fusion technology.Three MAbs were produced and characterized by indirect enzyme-linked immunosorbent assay(ELISA),western blotting,and immunofluorescence assay(IFA).Wes-tern blotting result showed that the MAbs could specifically react with the capsid protein of RGNNV.The result of IFA showed that the MAbs could recognize virions in RGNNV-infected grouper spleen(GS)cells.These results indicate that the MAbs can specifi-cally recognize RGNNV virions and can be used to produce a rapid detection method.This study provides a foundation for further studies on the rapid diagnosis of RGNNV and its infection mechanisms.展开更多
The origin of T cells in the teleost's brain is unclear.While viewing the central nervous system(CNS)as immune privileged has been widely accepted,previous studies suggest that T cells residing in the thymus but n...The origin of T cells in the teleost's brain is unclear.While viewing the central nervous system(CNS)as immune privileged has been widely accepted,previous studies suggest that T cells residing in the thymus but not in the spleen of the teleost play an essential role in communicating with the peripheral organs.Here,we identified nine T cell subpopulations in the thymus and spleen of orange-spotted grouper(Epinephelus coioices)through single-cell RNA-sequencing analysis.After viral CNS infection with red-spotted grouper nervous necrosis virus(RGNNV),the number of slc43a2^(+)T cells synchronously increased in the spleen and brain.During the infection tests in asplenic zebrafish(tlx1^▲zebrafish model),no increase in the number of slc43a2^(+)T cells was observed in the brain.Single-cell transcriptomic analysis indicated that slc43a2^(+)T cells mature and functionally differentiate within the spleen and then migrate into the brain to trigger an immune response.This study suggests a novel route for T cell migration from the spleen to the brain during viral infection in fish.展开更多
基金supported by the National Natural Science Foundation of China (No. 31972834)the National Key Research and Development Program of China (No. 2018YFD0900505)
文摘Nervous necrosis virus(NNV)can infect more than 120 fish species worldwide and has caused high mortality and sig-nificant economic losses to the aquaculture industry.Among different genotypes of NNV,the red-grouper nervous necrosis virus(RGNNV)is the most widely distributed one with the highest number of susceptible fish species.In this study,the capsid protein(Cp)gene of RGNNV was recombined and expressed in Escherichia coli strain BL21(DE3)and the recombinant Cp(rCp)was used as an immunogen to produce monoclonal antibodies(MAbs)through hybridoma cell fusion technology.Three MAbs were produced and characterized by indirect enzyme-linked immunosorbent assay(ELISA),western blotting,and immunofluorescence assay(IFA).Wes-tern blotting result showed that the MAbs could specifically react with the capsid protein of RGNNV.The result of IFA showed that the MAbs could recognize virions in RGNNV-infected grouper spleen(GS)cells.These results indicate that the MAbs can specifi-cally recognize RGNNV virions and can be used to produce a rapid detection method.This study provides a foundation for further studies on the rapid diagnosis of RGNNV and its infection mechanisms.
基金supported by the National Key Research and Development Program of China(2022YFD2400502)the National Natural Science Foundation of China(42176103,41825013,42230409,42276127)+2 种基金the Guangdong Provincial Natural Science Foundation(2022A1515012505)the Key-Area Research and Development Program of Guangdong Province(2021B0202040002)China Agricultural Research System(CARS-47-G16)。
文摘The origin of T cells in the teleost's brain is unclear.While viewing the central nervous system(CNS)as immune privileged has been widely accepted,previous studies suggest that T cells residing in the thymus but not in the spleen of the teleost play an essential role in communicating with the peripheral organs.Here,we identified nine T cell subpopulations in the thymus and spleen of orange-spotted grouper(Epinephelus coioices)through single-cell RNA-sequencing analysis.After viral CNS infection with red-spotted grouper nervous necrosis virus(RGNNV),the number of slc43a2^(+)T cells synchronously increased in the spleen and brain.During the infection tests in asplenic zebrafish(tlx1^▲zebrafish model),no increase in the number of slc43a2^(+)T cells was observed in the brain.Single-cell transcriptomic analysis indicated that slc43a2^(+)T cells mature and functionally differentiate within the spleen and then migrate into the brain to trigger an immune response.This study suggests a novel route for T cell migration from the spleen to the brain during viral infection in fish.
