目的:观察苦参素(MAT)对实验性自身免疫性脑脊髓炎(EAE)小鼠脊髓中Th1/Th17细胞表达的影响,探讨MAT对EAE小鼠的防治机制。方法:用MOG35-55诱导制备EAE小鼠模型,40只C57BL/6小鼠随机分为EAE组及MAT组(MAT,200mg/kg),MAT组从免疫后第13天...目的:观察苦参素(MAT)对实验性自身免疫性脑脊髓炎(EAE)小鼠脊髓中Th1/Th17细胞表达的影响,探讨MAT对EAE小鼠的防治机制。方法:用MOG35-55诱导制备EAE小鼠模型,40只C57BL/6小鼠随机分为EAE组及MAT组(MAT,200mg/kg),MAT组从免疫后第13天开始连续腹腔注射给药至第22天,同时EAE组小鼠腹腔注射等量0.9%氯化钠溶液,观察记录小鼠体质量变化,神经功能学评分,苏木素-伊红(HE)和Luxol Fast Blue(LFB)髓鞘染色观察病理改变,免疫荧光双染检测CD4+IFN-γ+与CD4+IL-17+共表达情况,Western Blot法测定IL-12蛋白表达情况。结果:与EAE组小鼠比较,MAT组显著降低EAE小鼠的神经功能学评分(P<0.01),减轻EAE小鼠神经炎症浸润(P<0.01)和髓鞘脱失(P<0.01),下调脊髓中CD4+IFN-γ+(P<0.05)与CD4+IL-17+(P<0.05)的共表达,且两组CD4+IL-17+与CD4+IFN-γ+的共表达均成正相关(r=0.888~0.933,P<0.05),同时降低脊髓中IL-12的蛋白表达(P<0.05)。结论:MAT对EAE小鼠具有防治作用,可能与下调小鼠脊髓中Th1/Th17细胞的表达有关。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
文摘目的:观察苦参素(MAT)对实验性自身免疫性脑脊髓炎(EAE)小鼠脊髓中Th1/Th17细胞表达的影响,探讨MAT对EAE小鼠的防治机制。方法:用MOG35-55诱导制备EAE小鼠模型,40只C57BL/6小鼠随机分为EAE组及MAT组(MAT,200mg/kg),MAT组从免疫后第13天开始连续腹腔注射给药至第22天,同时EAE组小鼠腹腔注射等量0.9%氯化钠溶液,观察记录小鼠体质量变化,神经功能学评分,苏木素-伊红(HE)和Luxol Fast Blue(LFB)髓鞘染色观察病理改变,免疫荧光双染检测CD4+IFN-γ+与CD4+IL-17+共表达情况,Western Blot法测定IL-12蛋白表达情况。结果:与EAE组小鼠比较,MAT组显著降低EAE小鼠的神经功能学评分(P<0.01),减轻EAE小鼠神经炎症浸润(P<0.01)和髓鞘脱失(P<0.01),下调脊髓中CD4+IFN-γ+(P<0.05)与CD4+IL-17+(P<0.05)的共表达,且两组CD4+IL-17+与CD4+IFN-γ+的共表达均成正相关(r=0.888~0.933,P<0.05),同时降低脊髓中IL-12的蛋白表达(P<0.05)。结论:MAT对EAE小鼠具有防治作用,可能与下调小鼠脊髓中Th1/Th17细胞的表达有关。
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.
