Morphine and gp120 modulation of Fc gamma mediated macrophage phagocytosis

说明(由申请人提供)：阿片类药物的使用和滥用已知会抑制一些免疫反应，因此，被认为是艾滋病毒-1感染进展的辅助因素。根据疾控中心的最新统计，自艾滋病毒流行以来，注射吸毒已直接和间接占美国艾滋病病例的三分之一以上(36%)。在HIV阳性患者中观察到的细菌败血症的高频率已被证明部分是由于先天免疫功能受损，特别是巨噬细胞功能(趋化、杀菌、吞噬和超氧化物产生)。我们的目标是更好地了解在存在HIV-1包膜蛋白gp120的情况下阿片类药物对先天免疫的影响。 这项提案的具体目标将集中在专门研究吗啡和gp120诱导抑制巨噬细胞内化和消除细菌感染能力的机制的实验上。巨噬细胞系以及体内和体外处理的原代腹膜巨噬细胞将被用来显示慢性吗啡和gp120对这一天然免疫成分的影响。目的一，研究慢性吗啡和gp120在体内和体外对Fc-γ受体(FcgR)介导的免疫球蛋白调理细菌颗粒吞噬功能的影响。我们将研究吗啡和gp120在调节cAMP、FcgR表达和激活以及肌动蛋白聚合中的作用。此外，在第二个目标中，我们建议研究慢性吗啡和gp120对吞噬-溶酶体融合的影响和细菌杀灭的机制。重点研究巨噬细胞释放活性氧中间体(ROI)、活性氮中间体(RNIs)和整体杀菌能力。此外，我们还将研究gp120和IF的作用以及它是如何调节吗啡诱导的细菌杀灭的。通过对吗啡和gp120存在下的吞噬和杀菌活性的详细分析，我们希望深入了解阿片类药物在HIV感染过程中对巨噬细胞清除细菌的一般影响。与公共卫生相关：艾滋病毒通过攻击负责清除细菌的免疫细胞，削弱了人体抗击疾病的能力。在免疫系统正常的人中罕见的感染对艾滋病毒携带者来说是致命的。在存在阿片类药物等滥用药物的情况下，这种影响进一步加剧。艾滋病毒与滥用药物之间的相互作用机制尚未得到很好的探讨。研究阿片类药物和HIV诱导免疫抑制的机制，对于设计更好的治疗方法，提高患者的生活质量具有重要意义。

DESCRIPTION (provided by applicant): Opiate use and abuse has been known to suppress a number of immune responses and, therefore, have been postulated to serve as cofactor in the progression of HIV-1 infection. According to latest CDC statistics, since the HIV epidemic began, injection drug use has directly and indirectly accounted for more than one-third (36%) of AIDS cases in the United States. The high frequency of bacterial sepsis observed in HIV-positive patients has been shown to be in part due to impairment of innate immunity, specifically macrophage function (chemotaxis, bacterial killing, phagocytosis, and superoxide production). Our goal is to better understand effects of opioids on innate immunity in presence of HIV-1 envelope protein gp120. The specific aims of this proposal will focus on experiments that specifically address mechanisms of morphine and gp120 induced suppression of macrophage's ability to internalize, and eliminate bacterial infections. Macrophage cell lines as well as primary peritoneal macrophages treated in vivo and in vitro will e used to show the effects of chronic morphine and gp120 on this constituent of innate immunity. In aim one, we intend to investigate in vivo and in vitro effects of chronic morphine and gp120 on Fc-gamma receptor (FcgR) mediated phagocytosis of IgG opsonized bacterial particles. We will examine the role of morphine and gp120 in modulation of cAMP, FcgR expression and activation as well as actin polymerization. Additionally in aim two, we propose to study the effects of chronic morphine and gp120 on phago-lysosomal fusion and mechanisms of bacterial killing. Focusing on release of reactive oxygen intermediates (ROIs), reactive nitrogen intermediates (RNIs) and overall bactericidal ability of macrophages. In addition, we will study the effects of gp120 and if and how it modulates morphine induced inhibition of bacterial killing. By performing a detailed analysis of phagocytosis and bactericidal activity in presence of morphine and gp120 we hope to gain insight in general effect of opiates on bacterial clearance by macrophages during HIV infection. PUBLIC HEALTH RELEVANCE: HIV weakens the body's ability to fight disease by targeting immune cells in charge of bacterial clearance. Infections which are rarely seen in those with normal immune systems are deadly to those with HIV. In presence of drugs of abuse such as opioids, this effect is further exacerbated. Mechanisms of interaction between HIV and drugs of abuse have not been well explored. It is important to study the mechanisms of opioid and HIV induced immune suppression so we can design better therapies and improve quality of life.