MOLECULAR MECHANISM OF VIRUS NEUTRALIZATION

长期目标是研究病毒的结构 糖蛋白及其参与的附着、穿透和 并确定它们如何与抗体分子相互作用以 中和病毒的传染性。尽管病毒的中和一直是 经过多年的经验性研究，人们对这种分子知之甚少 这一重要的自然宿主对病毒感染的反应是基础。 了解这一点是了解传染性的先决条件。 病毒-抗体复合体在体内产生以及它们如何在 许多病毒感染的疾病过程。 La Crosse是一种具有重要医学意义的病毒，已被选为研究的模型 抗体中和的机制。它的体内有两种糖蛋白 包膜，但其中只有一个，G1，与抗体有关 中和。G1将通过肽图谱进行表征，以确定 糖基化、二硫键等部位。单抗有 已经准备好并用于绘制G1上八个不重叠的抗原点。 这些抗原点的确切位置将通过使用 免疫沉淀G1的CNBR多肽片段的单特异性抗体， 对每个肽的氨基末端进行测序，并将其与 推导出的G1的氨基酸序列与他人提供的一致。它是 提出可能有一种主要的中和机制，但 在空斑分析中中和和测量的病毒数量可能是 每个病毒-抗体复合体被抑制的概率之和 在某种程度上与细胞相互作用。要确定这一点，必须有更多 了解La Crosse病毒的传染性进入。这件事会做到的 在存在或不存在的情况下使用结合穿透和融合分析 溶酶促进剂，如氯喹。病毒对每个人都有反应 单抗将在这些化验中进行检测，以确定 抗体必须结合在G1上才能中和病毒并确定病毒的作用部位 步骤(S)在繁殖周期中，病毒复制被阻止。这个 在转录时或之前发生中和的可能性也将 接受检查。有人提出，结构中的构象变化 G1是由抗体结合诱导的，这对病毒是必要的 中和。激光拉曼光谱将被用于尝试 确定是抗体结合还是抗体对的协同结合 在通过病毒传播的G1中诱导构象变化 薄膜。

The long-term objectives are to investigate the structure of virus glycoproteins and their involvement in attachment, penetration, and uncoating and to determine how they interact with antibody molecules to neutralize virus infectivity. Altough neutralization of virus has been studied emperically for many years, little is known about the molecular basis for this important natural host response to virus infection. Knowledge of this is a prerequisite to the understanding of how infectious virus-antibody complexes arise in vivo and how they contribute to the disease process of many viral infections. La Crosse, a medically important virus, has been chosen as a model to study the mechanism of antibody neutralization. It has two glycoproteins in its envelope, but only one of them, G1, is involved in antibody neutralization. G1 will be characterized by peptide mapping to determine sites of glycosylation, disulfide bonding, etc. Monoclonal antibodies have been prepared and used to map eight non-overlapping antigenic sites on G1. The exact location of these antigenic sites will be determined by using the monospecific antibodies to immunoprecipitate CNBr peptide fragments of G1, sequencing the amino terminus of each peptide, and correlating this with the deduced amino acid sequence of G1 as provided by others. It is proposed that there may be one main mechanism of neutralization but that the amount of virus neutralized and measured in plaque assays may be the sum of the probabilities that each virus-antibody complex will be inhibited from interacting with the cell at some level. To determine this, more must be known about the infectious entry of La Crosse virus. This will be done using binding penetration, and fusion assays in the presence or absence of lysosomotropic agents such as chloroquine. Virus reacted with each monoclonal antibody will be examined in these assays to determine where antibody must bind on G1 to neutralize virus and to determine at what step(s) in the multiplication cycle the virus replication is blocked. The possibility of neutralization at or just prior to transcription will also be examined. It is proposed that a conformational change in the structure of G1 is induced by antibody binding and that this is necessary for virus neutralization. Laser Raman spectroscopy will be used in attempts to determine if binding of antibody or synergistic binding of antibody pairs induces a conformational changes in G1 that is transmitted across the viral membrane.