EX VIVO EFFECT OF METHADONE USE AND WITHDRAWAL ON IMMUNE FUCTION AND HIV-1

美沙酮使用和戒断对免疫功能和 HIV-1 的体外影响

基本信息

批准号：
8573406
负责人：
JOSE W RODRIGUEZ
金额：
$ 12.24万
依托单位：
UNIVERSIDAD CENTRAL DEL CARIBE
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至 2017-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8573406
关键词：
Acquired Immunodeficiency Syndrome Acute Affect Animal Model Antibodies Biological Assay Biomedical Research CCL2 gene CD4 Positive T Lymphocytes CXCL10 gene CXCR4 Receptors Cell physiology Cells Cellular Immunity Chronic Clinical Color Data Disease Progression Drug abuse Enzyme-Linked Immunosorbent Assay Epidemiologic Studies Epidemiology Flow Cytometry Future Gene Expression HIV Core Protein p24 HIV-1 Human Immune Immune system In Vitro Individual Infection Instruction Interferons Interleukin-10 Interleukin-17 Interleukin-2 Interleukin-4 Interleukin-6 Intervention Laboratories Lead Longitudinal Studies MAPK14 gene MAPK8 gene Macrophage Inflammatory Protein-1 Mediating Methadone Molecular Opiate Addiction Opiates Opioid Receptor Patients Pharmaceutical Preparations Pilot Projects Population RANTES RNA Reverse Transcriptase Polymerase Chain Reaction Risk Factors Sum System TNF gene Testing Viral Virus Western Blotting Withdrawal base chemokine cofactor cytokine design drug abuser effective therapy extracellular healthy volunteer immune function in vivo mu opioid receptors protein expression receptor expression research study

项目摘要

Ex Vivo Effect of Methadone Use and Withdrawal on Immune Function and HIV-1 Replication of CD4+ T-Cells from Chronic Methadone Patients Drug abuse remains one of the major risk factors for the spread of HIV-1 infection with opiates being a principal drug of concern. Opiates have been shown to be determining factors in the rate of progression of AIDS. The sum of various in vitro, animal-model, and epidemiological studies Indicates that opiates have conditionally variable effects on HIV-1 disease progression. Immunovirological and pharmacological variability are two major factors that seem to dictate whether HIV-1 infection will progress more rapidly or more slowly in HIV-1 infected opiate-exposed individuals. According to previous studies, the immunovirological effect of an opiate drug such as methadone Is not well characterized in an ex vivo human system. The hypothesis to be tested in this proposal is that ex vivo methadone use and withdrawal will enhance HIV-1 infectivity in CD4+ T-lymphocytes by altering cytokine/chemokine profile and upregulating HIV-1 co-receptor and mu-opioid receptor expression. The specific aims will analyze various immunovirological determinants of HIV-1 infectivity to evaluate the effects of methadone use and withdrawal. CD4+ T-lymphocytes will be obtained from HIV-1 negative healthy volunteers and chronic (more than one year) methadone patients. Cells will be acutely infected with HIV-1 SF2 (X4/R5 dual virus strain) and then assayed to determine how ex vivo methadone exposure (10-7 and 10-6 M) and abrupt withdrawal: (1) affects HIV-1 infectivity (using HIV-1 p24 ELISA); (2) disrupt cytokine/chemokine profile (using cytometric bead array and ELISA); and (3) modulates the gene and protein expression of extracellular and intracellular molecules (using flow cytometry, quantitative RT-PCR, and Western blot). Statistical analysis will be used to determine significant differences between the different experimental conditions. This study will reveal new cellular and molecular mechanisms involved in methadone-mediated HIV-1 replication enhancement. This data can be used to design future in vivo longitudinal studies on the immune and viral modulating effects of methadone in HIV-1 infected methadone patients

美沙酮使用和戒断对来自慢性美沙酮患者的CD4+ T细胞的免疫功能和HIV-1复制的体内效应仍然是药物滥用的主要危险因素之一是HIV-1感染与阿片类药物的传播是主要关注的药物。鸦片蛋白酶已被证明是确定艾滋病进展速率的因素。各种体外，动物模型和流行病学研究的总和表明，阿片类药物对HIV-1疾病进展的有条件影响。免疫病毒学和药理变异性是两个主要因素，似乎决定了HIV-1感染在受HIV-1感染的鸦片暴露的个体中的进展更快还是更慢。根据先前的研究，在体内人类系统中，阿片类药物（如美沙酮）的免疫病毒学作用没有很好地表征。该提案中要检验的假设是，通过改变细胞因子/趋化因子谱并上调HIV-1共肽和MU-阿片受体表达，可以通过改变细胞因子/趋化因子谱和上调HIV-1的HIV-1共同因子谱，从而提高CD4+ T淋巴细胞中的HIV-1感染性。具体目的将分析HIV-1感染性的各种免疫病毒学决定因素，以评估美沙酮使用和戒断的影响。 CD4+ T淋巴细胞将从HIV-1负健康志愿者和慢性（超过一年）美沙酮患者中获得。细胞将被HIV-1 SF2（X4/R5双病毒菌株）急性感染，然后测定以确定过体美沙酮暴露（10-7和10-6 m）和突然戒断：（1）影响HIV-1感染性（使用HIV-1 P24 ELISA）；（2）破坏细胞因子/趋化因子谱（使用细胞仪阵列和ELISA）；（3）调节细胞外分子和细胞内分子的基因和蛋白质表达（使用流式细胞仪，定量RT-PCR和Western blot）。统计分析将用于确定不同实验条件之间的显着差异。这项研究将揭示与美沙酮介导的HIV-1复制增强有关的新细胞和分子机制。这些数据可用于设计有关美沙酮在HIV-1感染美沙酮患者中的免疫和病毒调节作用的未来体内纵向研究