Artemisia annua,a traditional Chinese medicinal plant,remains the only plant source for artemisinin production,yet few genes have been identified to be involved in both the response to biotic stresses,such as pathogen...Artemisia annua,a traditional Chinese medicinal plant,remains the only plant source for artemisinin production,yet few genes have been identified to be involved in both the response to biotic stresses,such as pathogens,and artemisinin biosynthesis.Here,we isolated and identified the WRKY transcription factor(TF)AaWRKY17,which could significantly increase the artemisinin content and resistance to Pseudomonas syringae in A.annua.Yeast one-hybrid(Y1H),dual-luciferase(dual-LUC),and electrophoretic mobility shift assay(EMSA)results showed that AaWRKY17 directly bound to the W-box motifs in the promoter region of the artemisinin biosynthetic pathway gene amorpha-4,11-diene synthase(ADS)and promoted its expression.Real-time quantitative PCR(RT-qPCR)analysis revealed that the transcript levels of two defense marker genes,Pathogenesis-Related 5(PR5)and NDR1/HIN1-LIKE 10(NHL10),were greatly increased in AaWRKY17-overexpressing transgenic A.annua plants.Additionally,overexpression of AaWRKY17 in A.annua resulted in decreased susceptibility to P.syringae.These results indicated that AaWRKY17 acted as a positive regulator in response to P.syringae infection.Together,our findings demonstrated that the novel WRKY transcription factor AaWRKY17 could potentially be used in transgenic breeding to improve the content of artemisinin and pathogen tolerance in A.annua.展开更多
Age-associated changes in immune cells have been linked to an increased risk for infection.However,a global and detailed characterization of the changes that human circulating immune cells undergo with age is lacking....Age-associated changes in immune cells have been linked to an increased risk for infection.However,a global and detailed characterization of the changes that human circulating immune cells undergo with age is lacking.Here,we combined scRNA-seq,mass cytometry and sCATAC-seq to compare immune cell types in peripheral blood collected from young and old subjects and patients with COVID-19.We found that the immune cell landscape was reprogrammed with age and was characterized by T cell polarization from naive and memory cells to effector,cytotoxic,exhausted and reg-ulatory cells,along with increased late natural killer cells,age-associated B cells,inflammatory monocytes and age-associated dendritic cells.In addition,the expression of genes,which were implicated in coron-avirus susceptibility,was upregulated in a cell subtype-specific manner with age.Notably,COVID-19 promoted age-induced immune cell polarization and gene expression related to inflammation and cellular senes-cence.Therefore,these findings suggest that a dysreg-ulated immune system and increased gene expression associated with SARS-CoV-2 susceptibility may at least partially account for COVID-19 vulnerability in the elderly.展开更多
Aging-induced changes in the immune system are associated with a higher incidence of infection and vaccination failure.Lymph nodes,which filter the lymph to identify and fight infections,play a central role in this pr...Aging-induced changes in the immune system are associated with a higher incidence of infection and vaccination failure.Lymph nodes,which filter the lymph to identify and fight infections,play a central role in this process.However,careful characterization of the impact of aging on lymph nodes and associated autoimmune diseases is lacking.We combined single-cell RNA sequencing(scRNA-seq)with flow cytometry to delineate the immune cell atlas of cervical draining lymph nodes(CDLNs)of both young and old mice with or without experimental autoimmune uveitis(EAU).We found extensive and complicated changes in the cellular constituents of CDLNs during aging.When confronted with autoimmune challenges,old mice developed milder EAU compared to young mice.Within this EAU process,we highlighted that the pathogenicity of T helper 17 cells(Th17)was dampened,as shown by reduced GM-CSF secretion in old mice.The mitigated secretion of GMCSF contributed to alleviation of IL-23 secretion by antigen-presenting cells(APCs)and may,in turn,weaken APCs’effects on facilitating the pathogenicity of Th17 cells.Meanwhile,our study further unveiled that aging downregulated GM-CSF secretion through reducing both the transcript and protein levels of IL-23R in Th17 cells from CDLNs.Overall,aging altered immune cell responses,especially through toning down Th17 cells,counteracting EAU challenge in old mice.展开更多
基金This work was supported by the National Key R&D Program of China(2018YFA0900600)the Bill&Melinda Gates Foundation(OPP1199872 and INV-027291)+1 种基金SJTU Trans-med Awards Research(20190104)the SJTU Global Strategic Partnership Fund(2020 SJTU-CORNELL).
文摘Artemisia annua,a traditional Chinese medicinal plant,remains the only plant source for artemisinin production,yet few genes have been identified to be involved in both the response to biotic stresses,such as pathogens,and artemisinin biosynthesis.Here,we isolated and identified the WRKY transcription factor(TF)AaWRKY17,which could significantly increase the artemisinin content and resistance to Pseudomonas syringae in A.annua.Yeast one-hybrid(Y1H),dual-luciferase(dual-LUC),and electrophoretic mobility shift assay(EMSA)results showed that AaWRKY17 directly bound to the W-box motifs in the promoter region of the artemisinin biosynthetic pathway gene amorpha-4,11-diene synthase(ADS)and promoted its expression.Real-time quantitative PCR(RT-qPCR)analysis revealed that the transcript levels of two defense marker genes,Pathogenesis-Related 5(PR5)and NDR1/HIN1-LIKE 10(NHL10),were greatly increased in AaWRKY17-overexpressing transgenic A.annua plants.Additionally,overexpression of AaWRKY17 in A.annua resulted in decreased susceptibility to P.syringae.These results indicated that AaWRKY17 acted as a positive regulator in response to P.syringae infection.Together,our findings demonstrated that the novel WRKY transcription factor AaWRKY17 could potentially be used in transgenic breeding to improve the content of artemisinin and pathogen tolerance in A.annua.
基金This work was supported by the National Key Research and Development Program of China(2017YFA0105804)the Strategic Priority Research Program of the Chinese Academy of Sciences(XDA16010000)+8 种基金the National Key Research and Development Program of China(2018YFC2000100,2017YFA0103304,2017YFA0102802,2018YFA0107203)the National Natural Science Foundation of China(81670897,81625009,91749202.81861168034,81921006,31671429,91949209,91749123,81671377,81822018,81870228,81922027,81701388,81601233)the Program of the Beijing Municipal Science and Technology Commission(Z191100001519005)Bejing Natural Science Foun-dation(Z190019)Bejing Municipal Commission of Health and Family Planning(PXM2018026283_000002)Advanced Innovation Center for Human Brain Protection(3500-1192012)the Key Research Program of the Chinese Academy of Sciences(KFZD-SW-221)K.C.Wong Education Foundation(GJTD-2019-06,GJTD-2019-08),Youth Innovation Promotion Association of CAS(2016093)the State Key Laboratory of Membrane Biology and the State Key Laboratory of Stem Cell and Reproductive Biology.
文摘Age-associated changes in immune cells have been linked to an increased risk for infection.However,a global and detailed characterization of the changes that human circulating immune cells undergo with age is lacking.Here,we combined scRNA-seq,mass cytometry and sCATAC-seq to compare immune cell types in peripheral blood collected from young and old subjects and patients with COVID-19.We found that the immune cell landscape was reprogrammed with age and was characterized by T cell polarization from naive and memory cells to effector,cytotoxic,exhausted and reg-ulatory cells,along with increased late natural killer cells,age-associated B cells,inflammatory monocytes and age-associated dendritic cells.In addition,the expression of genes,which were implicated in coron-avirus susceptibility,was upregulated in a cell subtype-specific manner with age.Notably,COVID-19 promoted age-induced immune cell polarization and gene expression related to inflammation and cellular senes-cence.Therefore,these findings suggest that a dysreg-ulated immune system and increased gene expression associated with SARS-CoV-2 susceptibility may at least partially account for COVID-19 vulnerability in the elderly.
基金the National Key Research and Development Program of China(2017YFA0105804)the Strategic Priority Research Program of the Chinese Academy of Sciences(XDA16010000)+3 种基金the National Key Research and Development Program of China(2020YFA0804000,2018YFC2000100,2017YFA0103304,2020YFA0803401,2019YFA0802202)the National Natural Science Foundation of China(Grant Nos.81921006,81625009,91749202,91949209,81822018,82125011,82122024,31970597)the Key Research Program of the Chinese Academy of Sciences(KFZD-SW-221)the 14th Five-year Network Security and Informatization Plan of Chinese Academy of Sciences(WX145XQ07-18)。
文摘Aging-induced changes in the immune system are associated with a higher incidence of infection and vaccination failure.Lymph nodes,which filter the lymph to identify and fight infections,play a central role in this process.However,careful characterization of the impact of aging on lymph nodes and associated autoimmune diseases is lacking.We combined single-cell RNA sequencing(scRNA-seq)with flow cytometry to delineate the immune cell atlas of cervical draining lymph nodes(CDLNs)of both young and old mice with or without experimental autoimmune uveitis(EAU).We found extensive and complicated changes in the cellular constituents of CDLNs during aging.When confronted with autoimmune challenges,old mice developed milder EAU compared to young mice.Within this EAU process,we highlighted that the pathogenicity of T helper 17 cells(Th17)was dampened,as shown by reduced GM-CSF secretion in old mice.The mitigated secretion of GMCSF contributed to alleviation of IL-23 secretion by antigen-presenting cells(APCs)and may,in turn,weaken APCs’effects on facilitating the pathogenicity of Th17 cells.Meanwhile,our study further unveiled that aging downregulated GM-CSF secretion through reducing both the transcript and protein levels of IL-23R in Th17 cells from CDLNs.Overall,aging altered immune cell responses,especially through toning down Th17 cells,counteracting EAU challenge in old mice.
