Effects of human cytomegalovirus on monocyte survival and differentiation

描述（由申请人提供）：人类巨细胞病毒（HCMV）感染在免疫正常的个体中通常是无症状的，尽管HCMV是几种慢性炎症性疾病（包括动脉粥样硬化和炎症性肠病）病因学中的主要候选病毒。在免疫功能低下的个体中，如新生儿、艾滋病患者和移植受者，HCMV感染可导致急性多器官炎症，从而导致显著的发病率和死亡率。与HCMV感染相关的炎症性器官疾病是在无症状或有症状感染期间发生的系统性病毒传播和多器官部位感染的直接后果。单核细胞负责将病毒输送到组织中，并在炎症状态中发挥核心作用

DESCRIPTION (provided by applicant): Human cytomegalovirus (HCMV) infection is generally asymptomatic in immunocompetent individuals, although HCMV is a primary viral candidate in the etiology of several chronic inflammatory diseases including atherosclerosis and inflammatory bowel disease. In immunocompromised individuals, such as neonates, AIDS patients, and transplant recipients, HCMV infection can lead to acute multi-organ inflammation resulting in significant morbidity and mortality. Inflammatory organ diseases associated with a HCMV infection is a direct consequence of the systemic viral spread to and infection of multiple organ sites that occur during either asymptomatic or symptomatic infections. Monocytes are responsible for delivering the virus into tissues and play a central role in the inflammatory state of infected organs. However, because anti-apoptotic viral proteins are not expressed during the early stages of infection, it remains unclear how HCMV simultaneously promotes the survival and pro-inflammatory differentiation of these short-lived cells. We hypothesize that HCMV stimulates a unique cellular anti-apoptotic reprogramming specifically designed to meet the viability needs of differentiating infected monocytes, which will be tested in 3 separate aims. Aim 1 will delineate which viral glycoprotein and cellular receptor interactions occurring during viral entry are responsible for the rapid block of monocyte apoptosis. Neutralizing antibodies and pharmaceutical inhibitors will be used individually or in different combinations to dissect the rol of each viral ligand and cellular receptor interaction in promoting the survival of HCMV-infected monocytes. Aim 2 will determine how HCMV concurrently drives the long-term survival and the pro-inflammatory differentiation of infected monocytes. Mutant viruses and anti-miRs [bind and inhibit microRNAs (miRNAs)] will be used to neutralize the activity of viral and cellular miRNAs during infection of monocytes. Monocyte survival and differentiation will then be examined by Annexin V staining and Affymetrix microarray gene analysis, respectively. Aim 3 will identify the molecular mechanism of action and therapeutic potential of targeting cellular anti-apoptotic Bcl-2 family proteins induced during HCMV infection of monocytes. To define the mode of action of virally induced cellular anti-apoptotic proteins, co-immunoprecipitation experiments will be done along with mass spectrometry to globally identify new and unique protein-protein interactions specific to HCMV-infected monocytes. BH3 profiling, a new technique used for predicting the sensitivity of cells to small-molecule inhibitors against anti-apoptotic Bcl-2 family members, will be used to determine the antiviral potential of targeting cellular Bcl-2 family survival proteins i order to direct infected monocytes towards cell death. These studies will increase our understanding on the pathogenesis and dissemination of HCMV, evaluate new antiviral strategies that target infected cells rather than the virus directly, and provide a proof-of-princile for the use of BH3 profiling to predict the efficacy of Bcl- 2 family small-molecule antagonists at eliminating specific virus and cell infection combinations.