Modulation of Monocyte Function by Cytomegalovirus IL-10

巨细胞病毒 IL-10 对单核细胞功能的调节

基本信息

批准号：
7031650
负责人：
JULIET VESCIO SPENCER
金额：
$ 6.52万
依托单位：
UNIVERSITY OF SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-04-01 至 2007-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7031650
关键词：
clinical research cytokine receptors cytomegalovirus gene expression human tissue immunoregulation interleukin 10 latent virus infection monocyte nuclear factor kappa beta protein kinase protein structure function receptor binding tissue /cell culture transcription factor virus infection mechanism

项目摘要

DESCRIPTION (provided by applicant): Cytomegalovirus is a widespread human pathogen with the ability to persist as a lifelong latent infection. Although CMV infection is typically subclinical, life-threatening disease may occur in the immunosuppressed. Our long-term goal is to identify mechanisms used by human CMV to establish latent infection and reactivate from latency. The hypothesis is that CMV-encoded interleukin-10 (cmvlL-10) signaling in monocytes triggers events that may facilitate establishment of or reactivation from latency. This is based on the following: 1) cmvlL-10 binds to the cellular IL-10 receptor (IL-10R), despite having only 27% sequence identity to human IL-10 (hlL-10), 2) cmvlL-10, like hlL-10, has potent immunosuppressive properties, such as down-regulation of MHC expression and inhibition of cytokine production by monocytes, and 3) cells harboring latent CMV express a transcript with homology to IL-10 encoding an alternately spliced form of cmvlL-10 (LA-cmvlL10). Furthermore, the UL111.5A region encoding cmvlL-10 is non-essential for lytic virus replication, suggesting a role for cmvlL-10 in other aspects of virus infection. Based on these observations, the proposed study will more thoroughly examine the effect of cmvlL-10 on monocytes, a site of virus persistence. The specific aims are: 1) Define signaling events that result from IL-10R engagement by cmv-lL10. We will examine a) activation of Stat transcription factors, b) participating protein kinases, and c) changes in gene expression. 2) Examine the mechanism for cmvlL-10-induced modulation of monocyte function. 1 possible mechanism involves inhibition of transcription factor NF-kappaB activity. To test this, we will examine a) Nf-kappaB activation, b) I-kappaB protein, levels, and c) I-kappaB-alpha phosphorylation state. 3) Determine whether LA-cmvlL-10 has biological function. We will a) express LA-cmvlL-10, b) examine binding to IL- 10R, and c) evaluate inhibition of cytokine production. The results will provide a better understanding of the molecular action of cmvlL-10, and possibly establish a role for cmvlL-10 in regulation of virus latency.

描述（由申请人提供）：巨细胞病毒是一种广泛传播的人类病原体，能够作为终生潜伏感染持续存在。虽然巨细胞病毒感染通常是亚临床的，但免疫抑制的人可能会发生危及生命的疾病。我们的长期目标是确定人类 CMV 用于建立潜伏感染并从潜伏状态重新激活的机制。假设单核细胞中 CMV 编码的白细胞介素 10 (cmvlL-10) 信号传导会触发可能促进潜伏期建立或重新激活的事件。这是基于以下内容：1) cmvlL-10 与细胞 IL-10 受体 (IL-10R) 结合，尽管与人 IL-10 (hIL-10) 仅具有 27% 的序列同一性，2) cmvlL-10，与 hIL-10 一样，具有有效的免疫抑制特性，例如下调 MHC 表达和抑制单核细胞产生细胞因子，以及 3) 含有潜在 CMV 的细胞表达与IL-10同源的转录物，编码交替剪接形式的cmvlL-10 (LA-cmvlL10)。此外，编码 cmvlL-10 的 UL111.5A 区域对于裂解病毒复制不是必需的，这表明 cmvlL-10 在病毒感染的其他方面发挥作用。基于这些观察结果，拟议的研究将更彻底地检查 cmvlL-10 对单核细胞（病毒持续存在的部位）的影响。具体目标是： 1) 定义由 cmv-lL10 参与 IL-10R 引起的信号传导事件。我们将检查 a) Stat 转录因子的激活，b) 参与的蛋白激酶，以及 c) 基因表达的变化。 2) 检查 cmvlL-10 诱导的单核细胞功能调节机制。一种可能的机制涉及抑制转录因子 NF-kappaB 活性。为了测试这一点，我们将检查 a) Nf-kappaB 激活，b) I-kappaB 蛋白水平，以及 c) I-kappaB-alpha 磷酸化状态。 3)确定LA-cmvlL-10是否具有生物学功能。我们将a)表达LA-cmvlL-10，b)检查与IL-10R的结合，以及c)评估细胞因子产生的抑制。这些结果将有助于更好地了解 cmvlL-10 的分子作用，并可能确定 cmvlL-10 在病毒潜伏期调节中的作用。