Morphine and gp120 modulation of Fc gamma mediated macrophage phagocytosis

描述（由申请方提供）：已知阿片类药物使用和滥用可抑制多种免疫应答，因此，假定其在HIV-1感染进展中起辅助因子作用。根据CDC的最新统计数据，自艾滋病开始流行以来，注射毒品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性脓毒症的高频率已被证明部分是由于先天免疫受损，特别是巨噬细胞功能（趋化性、细菌杀伤、吞噬作用和超氧化物产生）。我们的目标是更好地了解阿片类药物在HIV-1包膜蛋白gp 120存在下对先天免疫的影响。 本提案的具体目标将集中在专门解决吗啡和gp 120诱导抑制巨噬细胞内化能力和消除细菌感染的机制的实验上。巨噬细胞系以及在体内和体外处理的原代腹腔巨噬细胞将用于显示慢性吗啡和gp 120对先天免疫的这种成分的影响。目的一，我们打算在体内和体外研究慢性吗啡和gp 120对Fc-γ受体（FcgR）介导的IgG调理细菌颗粒的吞噬作用的影响。我们将研究吗啡和gp 120在调节cAMP、FcgR表达和激活以及肌动蛋白聚合中的作用。此外，在目标二中，我们计划研究慢性吗啡和gp 120对吞噬-溶酶体融合的影响以及细菌杀伤机制。重点关注活性氧中间体（ROI）、活性氮中间体（RNI）的释放和巨噬细胞的整体杀菌能力。此外，我们将研究gp 120的作用，以及它是否以及如何调节吗啡诱导的细菌杀伤抑制。通过对吗啡和gp 120存在下的吞噬作用和杀菌活性进行详细分析，我们希望了解阿片类药物在HIV感染期间对巨噬细胞细菌清除的一般影响。公共卫生相关性：艾滋病毒通过靶向负责细菌清除的免疫细胞来削弱人体对抗疾病的能力。在免疫系统正常的人中很少见到的感染对艾滋病毒感染者来说是致命的。在滥用药物如阿片类药物的情况下，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充分探讨。研究阿片类药物和HIV诱导的免疫抑制机制，对于设计更好的治疗方案和提高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.