Effect of alcohol and opioids on DC function in HCV and HIV infection

酒精和阿片类药物对 HCV 和 HIV 感染中 DC 功能的影响

• 批准号: 7555418
• 负责人: Nicole L Yonkers
• 金额: $ 4.1万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-25 至 2011-08-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7555418
• 关键词: Acute; Adam11 gene; Address; Affect; Alcohol consumption; Alcohols; Allogenic; Antigens; Autologous; Basic Science; Biological Assay; Blood specimen; CCL22 gene; CD4 Positive T Lymphocytes; Cells; Chronic; Clinical; Comorbidity; Defect; Disease; Disease Progression; Drug usage; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Frequencies; HIV; HIV Infections; Hepatitis C virus; High Prevalence; Immune; Impairment; In Vitro; Individual; Infection; Ligands; Liver diseases; Mediating; Methadone; Methodology; Mission; Morphine; National Institute of Drug Abuse; National Institute on Alcohol Abuse and Alcoholism; Natural Killer Cells; Numbers; Opioid; PDC gene; Pathogenesis; Pattern; Peripheral; Phenotype; Population; Reporting; Research; Sampling; Staging; Substance abuse problem; T-Cell Activation; T-Lymphocyte; alcohol effect; chlorambucil/dactinomycin/methotrexate protocol; chronic alcohol ingestion; cytokine; enzyme linked immunospot assay; immunoregulation; in vitro Assay; insight; interest; peripheral blood; phosducin; research study; response; therapy design; uptake

DESCRIPTION (provided by applicant): To address the specific mission interests of the NIAAA: "alcohol-related immune dysregulation in HIV with and without coinfection and other comorbidities" and NIDA basic science research: effects of drug use on immune correlates in HIV and co-infection, this proposal will evaluate alcohol and opioid affects on innate immune cell phenotype and function in the context of HCV, HIV, and HCV-HIV disease. SPECIFIC AIM 1: To determine the effect of chronic alcohol ingestion and opioid use on MDC and PDC numbers and function in HCV, HIV, and HCV/HIV infected hosts. METHODS: Non-expanded, immature MDC and PDC will be purified from HCV, HIV, and HCV-HIV infected hosts who are identified with either minimal to moderate, or heavy alcohol consumption, and identified as either receiving, or not receiving methadone. Healthy control samples without chronic alcohol ingestion or opioid use will be used as a reference. Cells will be analyzed for levels of costimulatory molecule expression, antigen uptake ability, and maturation in response to TLR ligand (flow cytometry and ELISA). In parallel, freshly purified and TLR stimulated MDC and PDC will be analyzed for ability to activate allogeneic healthy control naive CD4 T cells or autologous NK cells (cytokine ELISPOT). Clinical parameters will be used to evaluate associations between cellular defects and HCV or HIV disease activity/stage. SPECIFIC AIM 2: To determine the direct effect of in vitro alcohol and opioid exposure on healthy control, HCV, HIV and HCV/HIV MDC and PDC function. METHODS: Aim 2 methodology will be identical to that used in Aim 1 with the following exceptions (1)MDC and PDC will be isolated only from HCV, HIV, HCV/HIV infected and healthy control subjects who are not ingesting heavy amounts of alcohol or opioids, and (2)MDC and PDC will be pre-treated in vitro with alcohol and/or morphine, and washed prior to assay start. RATIONALE: There is a high prevalence of alcohol and opioid use within the HCV, HIV and HCV-HIV populations. Heavy alcohol consumption has been associated with enhanced liver disease in HCV infected individuals and increased disease progression in HIV infected individuals. Moreover, opioid use has been associated with increased HIV disease progression and opioid exposure with suppression of IFN-a mediated anti-HCV activity. Understanding the innate cellular immune modulations caused by both substance abuse in chronically infected individuals and/or acute alcohol or opioid exposure may provide insight into mechanisms of disease pathogenesis and facilitate proper treatment designs.

描述（由申请人提供）：为了解决 NIAAA 的特定使命利益：“有或没有合并感染和其他合并症的 HIV 中与酒精相关的免疫失调”和 NIDA 基础科学研究：药物使用对 HIV 和 co 免疫相关物的影响-感染，该提案将评估酒精和阿片类药物对 HCV、HIV 和 HCV-HIV 疾病背景下先天免疫细胞表型和功能的影响。具体目标 1：确定长期饮酒和阿片类药物使用对 HCV、HIV 和 HCV/HIV 感染宿主的 MDC 和 PDC 数量和功能的影响。方法：从 HCV、HIV 和 HCV-HIV 感染宿主中纯化未扩增的、未成熟的 MDC 和 PDC，这些宿主被确定为轻度至中度或大量饮酒，并被确定为接受或不接受美沙酮。未长期饮酒或使用阿片类药物的健康对照样本将用作参考。将分析细胞的共刺激分子表达水平、抗原摄取能力以及对 TLR 配体的成熟反应（流式细胞术和 ELISA）。同时，将分析新鲜纯化和 TLR 刺激的 MDC 和 PDC 激活同种异体健康对照初始 CD4 T 细胞或自体 NK 细胞（细胞因子 ELISPOT）的能力。临床参数将用于评估细胞缺陷与 HCV 或 HIV 疾病活动/阶段之间的关联。具体目标 2：确定体外酒精和阿片类药物暴露对健康对照、HCV、HIV 和 HCV/HIV MDC 和 PDC 功能的直接影响。方法：目标 2 的方法与目标 1 中使用的方法相同，但有以下例外： (1)MDC 和 PDC 仅从 HCV、HIV、HCV/HIV 感染者和健康对照受试者中分离，这些受试者没有摄入大量酒精或阿片类药物、(2)MDC 和 PDC 将在体外用酒精和/或吗啡进行预处理，并在测定开始前进行清洗。理由：HCV、HIV 和 HCV-HIV 人群中酒精和阿片类药物的使用率很高。大量饮酒与 HCV 感染者的肝病加重以及 HIV 感染者的疾病进展加剧有关。此外，阿片类药物的使用与 HIV 疾病进展的增加有关，而阿片类药物的暴露会抑制 IFN-α 介导的抗 HCV 活性。了解慢性感染者滥用药物和/或急性酒精或阿片类药物暴露引起的先天细胞免疫调节可能有助于深入了解疾病发病机制，并有助于正确的治疗设计。