MOLECULAR MECHANISM OF IFN-Y INDUCTION AND ACTIVITY

我们最近报道了两种EB病毒转化的人细胞系， 在暴露于热休克条件下，产生抗病毒活性。 这 活性已被表征为干扰素-γ（IFN-Gamma）。 之一 细胞系GM 3299组成性地产生高水平的IFN-α， 而GM 4408产生非常少（如果有的话）的IFN-α。 初始 对产生的IFN-α的表征表明， 这是一种不同于PHA刺激的T淋巴细胞的干扰素 IFN-γ。 在GM 3299中诱导的非常高水平的干扰素活性 可能反映了IFN-α和IFN-γ之间的增强或协同作用。 IFN-γ。 我们将纯化和表征这种“新型”干扰素。 我们 应详细描述诱导过程并克隆IFN-γ 基因 我们将尝试定义参与的核苷酸序列 “热感应。“使用克隆的EB病毒DNA片段，我们将检查 EB DNA合成和转录之间的关系，如果有的话， 干扰素诱导过程。 这些实验将在年内完成。 发热对干扰素诱导作用的体内研究。 我们测量了生殖器疱疹感染者的干扰素水平， 患者 在血清中检测到高水平的IFN-α， 在疱疹囊泡中。 PHA刺激的淋巴细胞产生10-100倍以上的 IFN-γ比刺激的对照个体的淋巴细胞。 违反 其它报道，IFN-γ抑制培养物中疱疹斑的形成。 IFN-α和IFN-β活性较低。 我们将检查 使用克隆的HSV基因的这种抗病毒活性的基础。 最近的报道表明，鼠IFN-γ与 巨噬细胞激活因子 将检查其是否存在人IFN-γ 使用单克隆抗体。 我们将尝试在功能上将 IFN-γ的三种功能：抗病毒，抗增殖， 巨噬细胞活化

We have recently reported that two EB virus-transformed human cell lines, on exposure to heat shock conditions, produce an antiviral activity. This activity has been characterized as interferon-Gamma (IFN-Gamma). One of the cell lines, GM3299, produces high levels of IFN-Alpha constitutively, whereas GM4408 produces very little (if any) IFN-Alpha. Initial characterization of the IFN-Alpha produced indicates the possibility that this is a species of interferon differing from PHA-stimulated T-lymphocyte IFN-Gamma. The very high levels of interferon activity induced in GM3299 by heat shock may reflect potentiation or synergism between IFN-Alpha and IFN-Gamma. We shall purify and characterize this 'novel' interferon. We shall characterize the induction process in detail and clone the IFN-Gamma gene. We shall attempt to define the nucleotide sequences involved in "heat-induction." Using cloned EB virus DNA segments, we shall examine the relationship, if any, between EB DNA synthesis and transcription and the interferon induction process. These experiments will be paralleled with in vivo studies on the effect of fever on interferon induction. We have measured the interferon levels in genital herpes-infected patients. High levels of IFN-Alpha are detected in the serum, and IFN-Beta in herpatic vesicles. PHA-stimulated lymphocytes produce 10-100 times more IFN-Gamma than stimulated lymphocytes of control individuals. Contrary to other reports, IFN-Gamma inhibits herpes plaque formation in culture. IFN-Alpha and IFN-Beta are less active. We shall examine the molecular basis of this antiviral activity of using cloned HSV genes. Recent reports indicate that murine IFN-Gamma is identical to macrophage-activating factor. This will be examined for human IFN-Gamma using monoclonal antibodies. We shall attempt to functionally separate the three functions of IFN-Gamma: antiviral, anti-proliferative, and macrophage activation.