目的:建立干扰素-(interferon-,IFN-)基因敲除小鼠慢性乙型肝炎病毒(hepatitis B virus,HBV)复制模型.方法:IFN-基因敲除(IFN--/-)小鼠繁育并抽提组织DNA进行聚合酶链反应(PCR)及凝胶电泳鉴定基因型.IFN--/-小鼠纯合子9只与野生型C57BL/...目的:建立干扰素-(interferon-,IFN-)基因敲除小鼠慢性乙型肝炎病毒(hepatitis B virus,HBV)复制模型.方法:IFN-基因敲除(IFN--/-)小鼠繁育并抽提组织DNA进行聚合酶链反应(PCR)及凝胶电泳鉴定基因型.IFN--/-小鼠纯合子9只与野生型C57BL/6小鼠9只同时高压水注射pAAV/HBV1.2质粒,按既定时间点采血检测乙型肝炎表面抗原(hepatitis B virus surface antigen,HBsAg)、乙型肝炎e抗原(hepatitis B virus e antigen,HBeAg)和HBV DNA.血清HBsAg和HBeAg表达水平由电化学发光法进行定量检测.经抽提血清总DNA后,血清HBV DNA由定量PCR进行检测.结果:本实验室繁殖的IFN--/-小鼠均为纯合子基因型.IFN--/-小鼠和野生型C57BL/6小鼠血清中HBsAg、HBeAg和HBV DNA持续存在,转染后第40天仍阳性.但是,IFN--/-小鼠血清HBsAg表达水平高于C57BL/6野生小鼠(40天时,P=0.042);IFN--/-小鼠血清HBV DNA持续高水平复制,明显高于C57BL/6野生小鼠(第25天时,P=0.012;第40天时,P=0.039).两组小鼠血清HBeAg表达水平无差异.结论:IFN--/-小鼠慢性HBV复制模型成功建立,并揭示了IFN-在慢性HBV感染中可抑制HBV复制.展开更多
Thichosanthin (TCS) is a potent allergen to mice. According to our previous experiments, it could bring out the IgE response to ovabumin (OVA) if TCS was given one day before OVA immunization, while OVA alone could no...Thichosanthin (TCS) is a potent allergen to mice. According to our previous experiments, it could bring out the IgE response to ovabumin (OVA) if TCS was given one day before OVA immunization, while OVA alone could not induce IgE to it. In this work, the kinetics of interleukin 4(IL-4) and interferon γ(IFN-γ) gene expression in the mesenteric lymph node (MLN) of TCS-immunized mice was investigated using a semi-quantitative RT-PCR method. It indicated that TCS induced significant IL-4gene expression and the peaks of IL4 gene expression were on day one after TCS immunization in both primary and secondary response. In contrast, the IFN-γ gene expression was suppressed. Furthermore, the IL-4 gene expression in the secondary response was lower than that in the primary response. Thus the presence of IgE rpemory B cells were studied. Results showed that the amount of mature IgE mRNA arose significantly and rapidly one day after TCS restimulation, while in the MLN of the mice primed 30 days before and without boost, it was almost as the same amount of the unimmunized control. These findings suggest the existence of the IgE memory B cells in the mice after the primary TCS immunization.展开更多
文摘目的:建立干扰素-(interferon-,IFN-)基因敲除小鼠慢性乙型肝炎病毒(hepatitis B virus,HBV)复制模型.方法:IFN-基因敲除(IFN--/-)小鼠繁育并抽提组织DNA进行聚合酶链反应(PCR)及凝胶电泳鉴定基因型.IFN--/-小鼠纯合子9只与野生型C57BL/6小鼠9只同时高压水注射pAAV/HBV1.2质粒,按既定时间点采血检测乙型肝炎表面抗原(hepatitis B virus surface antigen,HBsAg)、乙型肝炎e抗原(hepatitis B virus e antigen,HBeAg)和HBV DNA.血清HBsAg和HBeAg表达水平由电化学发光法进行定量检测.经抽提血清总DNA后,血清HBV DNA由定量PCR进行检测.结果:本实验室繁殖的IFN--/-小鼠均为纯合子基因型.IFN--/-小鼠和野生型C57BL/6小鼠血清中HBsAg、HBeAg和HBV DNA持续存在,转染后第40天仍阳性.但是,IFN--/-小鼠血清HBsAg表达水平高于C57BL/6野生小鼠(40天时,P=0.042);IFN--/-小鼠血清HBV DNA持续高水平复制,明显高于C57BL/6野生小鼠(第25天时,P=0.012;第40天时,P=0.039).两组小鼠血清HBeAg表达水平无差异.结论:IFN--/-小鼠慢性HBV复制模型成功建立,并揭示了IFN-在慢性HBV感染中可抑制HBV复制.
文摘Thichosanthin (TCS) is a potent allergen to mice. According to our previous experiments, it could bring out the IgE response to ovabumin (OVA) if TCS was given one day before OVA immunization, while OVA alone could not induce IgE to it. In this work, the kinetics of interleukin 4(IL-4) and interferon γ(IFN-γ) gene expression in the mesenteric lymph node (MLN) of TCS-immunized mice was investigated using a semi-quantitative RT-PCR method. It indicated that TCS induced significant IL-4gene expression and the peaks of IL4 gene expression were on day one after TCS immunization in both primary and secondary response. In contrast, the IFN-γ gene expression was suppressed. Furthermore, the IL-4 gene expression in the secondary response was lower than that in the primary response. Thus the presence of IgE rpemory B cells were studied. Results showed that the amount of mature IgE mRNA arose significantly and rapidly one day after TCS restimulation, while in the MLN of the mice primed 30 days before and without boost, it was almost as the same amount of the unimmunized control. These findings suggest the existence of the IgE memory B cells in the mice after the primary TCS immunization.
