Role Of Monocytes In AIDS And As Targets For Antiviral Therapy

单核细胞在艾滋病中的作用及其作为抗病毒治疗的靶标

基本信息

批准号：
7733907
负责人：
Sharon M Wahl
金额：
$ 116.35万
依托单位：
NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
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
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7733907
关键词：
ANXA2 gene Accounting Acids Acquired Immunodeficiency Syndrome Aftercare Anti-Retroviral Agents Antigens Antiviral Agents Antiviral Response Antiviral Therapy Binding Bioavailable Blood CDKN1A gene CXCR4 Receptors Cancer Patient Cells Clinical Research Complementary DNA Consequences of HIV Cytidine Deaminase Data Depth Disease Progression Employee Strikes Environment Epithelium Esters Exhibits Exposure to Family Family member Gene Expression Genes Genetic Transcription Goals HIV HIV Infections HIV-1 Highly Active Antiretroviral Therapy Human Immediate-Early Genes Immune Immunologic Deficiency Syndromes In Vitro Infection Inflammation Inflammatory Interferons Intervention Lesion Life Cycle Stages Ligands Link Lymphocyte Lymphoid Cell Lymphoid Tissue Molecular Target Mononuclear Mycobacterium avium National Institute of Allergy and Infectious Disease Nature Opportunistic Infections PPAR gamma Pathogenesis Pathway interactions Patients Pattern Peripheral Blood Mononuclear Cell Pharmaceutical Preparations Phase I Clinical Trials Phosphorylation Population Predisposition Protease Inhibitor Proteins Proteolysis Readiness Regulation Research Personnel Resources Role SLPI gene Signal Transduction Site Testing Tissues Tonsil Toxic effect Viral Viral Load result Virion Virus Week apolipoprotein B mRNA editing enzyme cDNA Arrays cellular targeting cytokine drug resistant virus in vitro Model in vivo interest kinase inhibitor laser capture microdissection macrophage member monocyte oncoprotein p21 oral HIV oral cavity epithelium permissiveness polypeptide prevent resistance factors transmission process

项目摘要

Innate defense against HIV Mucosal associated lymphoid tissues are major targets of HIV during early infection and disease progression, and can also provide a viral safe haven during highly active antiretroviral therapy. Among these tissues, the tonsils remain enigmatic regarding their status as primary and/or secondary sites of retroviral infection, particularly since oral HIV transmission appears rare. To dissect the mechanisms underlying accessibility and susceptibility to HIV in this compartment, we dissected the epithelium with laser capture microdissection (LCM) and performed microarray gene expression analyses. Our studies demonstrated that the tonsil epithelium is a unique site and exhibits heightened potential for HIV transmission compared to other oral epithelia, likely due in part, to increased HIV co-receptor CXCR4 and reduced antiviral SLPI. To identify selective factors that favor the tonsil lymphoid cells as a viral reservoir, we compared isolated tonsil cell susceptibility to that of blood lymphocytes. In these studies, we identified several unique aspects of the tonsil micromilieu that may support its permissive nature, including elevated levels of Th2 cytokines and limited Th1 cytokines compared with PBMC. Despite elevated levels of antiviral IFN, the tonsil was not able to mount an effective antiviral response after exposure to HIV in vitro, nor apparently in vivo, which may reflect constitutive expression of negative regulators of IFN signaling, such as SOCS. SOCS appear to contribute to the blockade of IFN signaling cascades in the tonsil, evident by the reduced phosphorylation of Stat1, which may account for the local disengagement of IFN signaling and reduced antiviral activity. In a cyclic pattern, SOCS3 may also promote Th2 polarization. Such a profile of immune regulation is consistent with the necessity to control immune activation, yet maintain a state of readiness in an environment constantly bombarded with antigens. In ongoing studies, the tonsil multinucleated and mononuclear cells that produce striking amounts of HIV are being isolated and studied, as is the potential involvement of Treg as regulators and targets for the virus. Further understanding of these populations and the immunoregulatory pathways that influence their susceptibility to HIV pathogenesis are important for protecting vulnerable mucosal compartments from serving as reservoirs for the virus. In earlier studies, we demonstrated that a synthetic triterpenoid and peroxisome proliferator-activated receptor gamma (PPARgamma) ligand, 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid (CDDO), known to influence p21 kinase inhibitor expression (linked to viral life cycle), suppressed viral replication in macrophages and PBMC. Recently, a new methyl ester derivative of CDDO (CDDO-Me), which is uniquely orally bioavailable, has been synthesized and in preliminary phase I trials in cancer patients, it has been shown to not have significant toxicities, consistent with the possibility of testing this agent in HIV patients. Of considerable interest is the evidence that this methyl ester derivative targets the tissues, rather than the blood, and therefore we have focused on defining its ability to inhibit HIV in tonsil tissue derived CD4+ HIV target cells. As anti-retroviral therapy is often characterized by high toxicity and frequently results in the emergence of drug resistant virus strains, the identification of new anti-viral agents targeting host cell molecules that can be used independently or in conjunction with current anti-viral drugs will provide additional resources in the treatment of HIV infection. In additional clinical studies, we have been collaborating with investigators at NIAID in the treatment of AIDS patients with IFN in an attempt to define the antiviral mechanisms by which IFN suppresses HIV burden. In concordance with our in vitro data that IFN potently upregulates members of the cytidine deaminase family of innate intracellular proteins, we show that this also occurs in the PBMC of treated patients. Apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like 3G (APOBEC3G), a cytidine deaminase with lethal activity against HIV is packaged into progeny virions and leads to HIV cDNA degradation. As a counterattack, HIV virion infectivity factor (Vif) targets APOBEC3G for proteasomal proteolysis, thus excluding its incorporation into budding virions. We found that IFN treatment increased APOBEC3G and other family members, while reducing patient viral load. The maximal magnitude of viral decline occurred during the first week after treatment, especially in patients with lower viral load, comparable to that of early protease inhibitors. Thus, further consideration of a role for IFN as an inductive therapy in the context of low viral loads may be warranted and additional clinical studies are in progress. Moreover, through the use of microarrays, we have identified additional molecular targets of IFN that may contribute to its antiviral effects and may be considered as additional intervention approaches.

对艾滋病毒的先天防御粘膜相关的淋巴组织是早期感染和疾病进展过程中HIV的主要靶标，并且在高度活跃的抗逆转录病毒疗法期间也可以提供病毒避风港。在这些组织中，扁桃体在其作为逆转录病毒感染的原发性和/或次要部位的地位仍然神秘，特别是因为口腔HIV传播似乎很少。为了剖析该隔室中可及性和对HIV的易感性的机制，我们用激光捕获显微解剖（LCM）解剖上皮，并进行了微阵列基因表达分析。我们的研究表明，扁桃体上皮是一个独特的部位，与其他口腔上皮相比（可能部分归因于HIV共受体CXCR4）和抗病毒SLPI的其他口腔上皮相比，HIV传播的潜力更高。为了确定有利于扁桃体淋巴样细胞的选择性因素作为病毒储存剂，我们将分离的扁桃体细胞易感性与血液淋巴细胞的敏感性进行了比较。在这些研究中，我们确定了扁桃体微米的几个独特方面，这些方面可能支持其允许性质，包括与PBMC相比，Th2细胞因子水平升高和Th1细胞因子有限。尽管抗病毒IFN水平升高，但扁桃体在体外暴露于HIV后也无法在体内或显然在体内安装有效的抗病毒反应，这可能反映了IFN信号传导的负调节剂（例如SOCS）的构成表达。 SOC似乎有助于扁桃体中IFN信号级联的阻塞，这可以证明STAT1的磷酸化降低，这可能解释了IFN信号传导的局部脱离和抗病毒活性的降低。在环状模式下，SOCS3也可能促进Th2极化。这种免疫调节的特征与控制免疫激活的必要性是一致的，但在不断被抗原轰炸的环境中保持准备状态。在正在进行的研究中，正在分离和研究扁桃体多核细胞和单核细胞，TREG作为病毒的调节剂和靶标的潜力也被分离和研究。对这些人群的进一步了解以及影响其易感性艾滋病毒发病机理的免疫调节途径对于保护脆弱的粘膜室免于作为病毒的储层很重要。在较早的研究中，我们证明了合成三萜和过氧化物酶体增殖物激活的受体γ（ppargamma）配体，2-钙芳-3,12-12-12-Dioxooxooleana-1,9--dien-28-28- oic酸（CDDO），已知会影响P21 Kinase抑制作用（抑制P21 KINase抑制剂），并在病毒酶抑制剂上抑制了病毒式抑制作用，并将其流动周期循环），并在内PBMC。最近，一种新的CDDO（CDDO-ME）的甲基酯衍生物是独特的口头生物利用物，已合成，并且在癌症患者的初步I期试验中，它已被证明没有明显的毒性，与在HIV患者中测试该药物的可能性一致。引人注目的有证据表明，这种甲基酯衍生物靶向组织，而不是血液，因此我们专注于定义其在扁桃体组织中抑制HIV的能力，这是扁桃体组织中的CD4+ HIV靶细胞。由于通常以高毒性为特征，并且经常导致抗药性病毒菌株的出现，因此鉴定靶向靶向宿主细胞分子的新的抗病毒剂，可以独立或与当前的抗病毒药物一起使用，将为艾滋病毒感染提供其他资源。在其他临床研究中，我们一直与NIAID的研究人员合作，以治疗IFN的IFN患者，以定义IFN抑制HIV负担的抗病毒机制。与我们的体外数据一致，这些数据不强大地上调了先天性细胞内蛋白的胞苷脱氨酶家族的成员，我们表明这也发生在治疗患者的PBMC中。载脂蛋白B mRNA编辑酶催化性多肽样3G（APOBEC3G），一种具有致命活性HIV活性的胞苷脱氨酶被包装到后代病毒体中，并导致HIV CDNA降解。作为反攻击，HIV病毒体感染因子（VIF）靶向蛋白酶体蛋白水解，因此将其掺入萌芽的病毒体中。我们发现IFN治疗增加了Apobec3g和其他家庭成员，同时减少了患者的病毒载量。病毒降低的最大幅度发生在治疗后的第一周，尤其是病毒载量较低的患者，与早期蛋白酶抑制剂相当。因此，在低病毒负荷的背景下，进一步考虑了IFN作为归纳疗法的作用，并且可能有其他临床研究正在进行中。此外，通过使用微阵列，我们已经确定了可能有助于其抗病毒作用的IFN的其他分子靶标，可以被视为其他干预方法。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Plasminogen activator inhibitor-2 (PAI-2) in eosinophilic leukocytes.

嗜酸性白细胞中的纤溶酶原激活剂抑制剂 2 (PAI-2)。

DOI：
10.1189/jlb.0304182
发表时间：
2004
期刊：
Journal of leukocyte biology
影响因子：
5.5
作者：
Swartz,JonathanM;Bystrom,Jonas;Dyer,KimberlyD;Nitto,Takeaki;Wynn,ThomasA;Rosenberg,HeleneF
通讯作者：
Rosenberg,HeleneF

Differential mucosal susceptibility in HIV-1 transmission and infection.

HIV-1 传播和感染的不同粘膜易感性。

DOI：
10.1177/154407370601900111
发表时间：
2006
期刊：
Advances in dental research
影响因子：
0
作者：
Moutsopoulos,NM;Greenwell-Wild,T;Wahl,SM
通讯作者：
Wahl,SM

Secretory leukocyte protease inhibitor binds to annexin II, a cofactor for macrophage HIV-1 infection.

DOI：
10.1084/jem.20041115
发表时间：
2004-11-15
期刊：
JOURNAL OF EXPERIMENTAL MEDICINE
影响因子：
15.3
作者：
Ma, G;Greenwell-Wild, T;Lei, KJ;Jin, WW;Swisher, J;Hardegen, N;Wild, CT;Wahl, SM
通讯作者：
Wahl, SM

Protein engineering of interferon alphas.

干扰素α的蛋白质工程。