Activation of inflammatory cells and upregulations of a number of cytokines in the central nervous system(CNS)of patients with prion diseases are frequently observed.To evaluate the potential changes of some brain cyt...Activation of inflammatory cells and upregulations of a number of cytokines in the central nervous system(CNS)of patients with prion diseases are frequently observed.To evaluate the potential changes of some brain cytokines that were rarely addressed during prion infection,the levels of 17 different cytokines in the brain homogenates of mice infected with different scrapie mouse-adapted agents were firstly screened with Luminex assay.Significant upregulations of interferon gamma-induced protein 10(IP10),keratinocyte chemoattractant(KC)and macrophage colony stimulating factor(M-CSF)were frequently detected in the brain lysates of many strains of scrapie infected mice.The upregulations of those three cytokines in the brains of scrapie infected mice were further validated by the individual specific ELISA and immunohistochemical assay.Increased specific mRNAs of IP10,M-CSF and KC in the brains of scrapie infected mice were also detected by the individual specific qRT-PCRs and IP10-specific digital PCR.Dynamic analyses of the brain samples collected at different time points post infection revealed the time-dependent increases of those three cytokines,particularly IP10 during the incubation period of scrapie infection.In addition,we also found that the levels of IP10 in cerebral spinalfluid(CSF)of 45 sporadic Creutzfeldt–Jakob disease(sCJD)patients were slightly but significantly higher than those of the cases who were excluded the diagnosis of prion diseases.These data give us a better understanding of inflammatory reaction during prion infection and progression of prion disease.展开更多
Objective To analyze the relationship between Chemokine IP10 and its receptor CXCR3 during prion infection.Methods We investigated the increases in IP10 signals,primarily localized in neurons within the brains of scra...Objective To analyze the relationship between Chemokine IP10 and its receptor CXCR3 during prion infection.Methods We investigated the increases in IP10 signals,primarily localized in neurons within the brains of scrapie-infected mice,using western blotting,ELISA,co-immunoprecipitation,immunohistochemistry,immunofluorescence assays,and RT-PCR.Results Both CXCR3 levels and activation were significantly higher in the brains of scrapie-infected mice and prion-infected SMB-S15 cells.Enhanced CXCR3 expression was predominantly observed in neurons and activated microglia.Morphological colocalization of PrPC/PrPSc with IP10/CXCR3 was observed in scrapie-infected mouse brains using immunohistochemistry and immunofluorescence.immunohistochemistry(IHC)analysis of whole brain sections further revealed increased accumulation of IP10/CXCR3 specifically in brain regions with higher levels of PrPSc deposits.Co-immunoprecipitation and biomolecular interaction assays revealed the molecular interactions between PrP and IP10/CXCR3.Notably,a significantly larger amount of IP10 accumulated within prion-infected SMB-S15 cells than in the normal partner cell line,SMB-PS.Importantly,resveratrol treatment effectively suppressed prion replication in SMB-S15 cells,thereby restoring the accumulation and secretion pattern of cellular IP10 similar to that observed in SMB-PS cells.Conclusion Our data demonstrate that the activation of IP10/CXCR3 signaling in prion-infected brain tissues coincides with PrPSc deposition.Modulation of IP10/CXCR3 signaling in the brain represents a potential therapeutic target for mitigating the progression of prion diseases.展开更多
基金supported by the National Natural Science Foundation of China(81772197,81401670 and 81630062)the Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences(2018RC330004)+3 种基金National Key R&D Program of China(2018YFC1200305Y)National Science and Technology Major Project of China(2018ZX10102001)SKLID Development Grant(2019SKLID401 and 2016SKLID603)the Natural Science Foundation of Heilongjiang Province(C2018044)
文摘Activation of inflammatory cells and upregulations of a number of cytokines in the central nervous system(CNS)of patients with prion diseases are frequently observed.To evaluate the potential changes of some brain cytokines that were rarely addressed during prion infection,the levels of 17 different cytokines in the brain homogenates of mice infected with different scrapie mouse-adapted agents were firstly screened with Luminex assay.Significant upregulations of interferon gamma-induced protein 10(IP10),keratinocyte chemoattractant(KC)and macrophage colony stimulating factor(M-CSF)were frequently detected in the brain lysates of many strains of scrapie infected mice.The upregulations of those three cytokines in the brains of scrapie infected mice were further validated by the individual specific ELISA and immunohistochemical assay.Increased specific mRNAs of IP10,M-CSF and KC in the brains of scrapie infected mice were also detected by the individual specific qRT-PCRs and IP10-specific digital PCR.Dynamic analyses of the brain samples collected at different time points post infection revealed the time-dependent increases of those three cytokines,particularly IP10 during the incubation period of scrapie infection.In addition,we also found that the levels of IP10 in cerebral spinalfluid(CSF)of 45 sporadic Creutzfeldt–Jakob disease(sCJD)patients were slightly but significantly higher than those of the cases who were excluded the diagnosis of prion diseases.These data give us a better understanding of inflammatory reaction during prion infection and progression of prion disease.
基金supported by SKLID Development Grants(2021SKLID504,2019SKLID401,2019SKLID603,and 2016SKLID603)the National Natural Science Foundation of China(81772197,81401670,and 81630062)。
文摘Objective To analyze the relationship between Chemokine IP10 and its receptor CXCR3 during prion infection.Methods We investigated the increases in IP10 signals,primarily localized in neurons within the brains of scrapie-infected mice,using western blotting,ELISA,co-immunoprecipitation,immunohistochemistry,immunofluorescence assays,and RT-PCR.Results Both CXCR3 levels and activation were significantly higher in the brains of scrapie-infected mice and prion-infected SMB-S15 cells.Enhanced CXCR3 expression was predominantly observed in neurons and activated microglia.Morphological colocalization of PrPC/PrPSc with IP10/CXCR3 was observed in scrapie-infected mouse brains using immunohistochemistry and immunofluorescence.immunohistochemistry(IHC)analysis of whole brain sections further revealed increased accumulation of IP10/CXCR3 specifically in brain regions with higher levels of PrPSc deposits.Co-immunoprecipitation and biomolecular interaction assays revealed the molecular interactions between PrP and IP10/CXCR3.Notably,a significantly larger amount of IP10 accumulated within prion-infected SMB-S15 cells than in the normal partner cell line,SMB-PS.Importantly,resveratrol treatment effectively suppressed prion replication in SMB-S15 cells,thereby restoring the accumulation and secretion pattern of cellular IP10 similar to that observed in SMB-PS cells.Conclusion Our data demonstrate that the activation of IP10/CXCR3 signaling in prion-infected brain tissues coincides with PrPSc deposition.Modulation of IP10/CXCR3 signaling in the brain represents a potential therapeutic target for mitigating the progression of prion diseases.
