Immunoregulation /Immune Recognition In Filarial/Nonfilarial Parasitic Infection

丝虫/非丝虫寄生虫感染中的免疫调节/免疫识别

基本信息

批准号：
9161431
负责人：
Thomas Nutman
金额：
$ 186.9万
依托单位：
NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
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/9161431
关键词：
70-kDa Ribosomal Protein S6 Kinases Acute Aerosols Agonist Antigen-Presenting Cells Antigens CCL11 gene CCR1 gene CD14 gene CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes CD8B1 gene Cell physiology Cells Characteristics Chronic Clinical Cyclophilins Data Dendritic Cells Development Disease Ecuador Epidemiology Eukaryotic Initiation Factor-2 FCGR3B gene Filarial Elephantiases Flow Cytometry Functional disorder Gene Expression Gene Expression Profile Gene Expression Profiling Genetic Goals Growth HIV Helminths Homologous Gene Human IRF1 gene Immune Immune response Immunity Immunologics In Vitro India Individual Infection Interferon Type II Interleukin-10 Interleukin-12 Intestines Legal patent Life Loiasis Lung Lymphatic Macrophage Activation Malaria Mali Mediating Memory Microfilaria Molecular Profiling Mus Mycobacterium tuberculosis Myelogenous Nematoda Onchocerciasis Outcome Parasite Control Parasites Parasitic infection Pathway interactions Phenotype Play Population Process Production Proliferating Regulation Regulatory T-Lymphocyte Research Role Signal Pathway Signal Transduction Stem cells System T cell response T memory cell T-Cell Proliferation T-Lymphocyte TLR3 gene TLR4 gene Transforming Growth Factor beta Tuberculosis Viral Tumor Antigens Virulent WISP3 gene Work anergy base chemotherapy co-infection cytokine effective therapy exposed human population gastrointestinal infection immunoregulation in vivo Model interleukin-19 liquid chromatography mass spectrometry mTOR protein macrophage member monocyte mycobacterial prevent programs protein expression response transcription factor

项目摘要

The hallmark of the immune response seen in individuals with patent lymphatic filariasis is a profound inability to proliferate or produce cytokines associated with a Type 1 response (IL 2 and IFN-gamma;) in response to parasite antigen. This parasite specific anergy is mediated, in large part, by IL 10 with TGF-beta; and CTLA-4 playing smaller regulatory roles. Other members of the IL-10 superfamily (IL-19 and IL-24) have been now shown to be upregulated in patent LF, a process driven by IL-10 itself. The mechanisms underlying the modulation of parasite-specific responses in LF continues to be a major research question. Using multiparameter flow cytometry, this antigen-specific modulation was shown to be associated with an expansion of IL-10 producing adaptive Treg and altered development of parasite-specific central and effector memory T cell populations. We have demonstrated that these Tregs influence both effector T cells and antigen presenting cells (APCs), but alter APC function indirectly through the effector T cells (Metenou et al, submitted). Most recently, we have identified, using protein expression profiling (by LC/MS/MS) of nTregs in the context of filarial infection, 2 additional molecules (WISP3 and PSG-1) that are secreted by nTregs that suppress T cell proliferation and IFN-gamma production by effector T cells in a contact independent manner. We have also shown that these nTregs in filarial infection are highly heterogeneous and activated but are programmed to be short lived. This may underlie their increase in number that is characteristic of infection with filarial parasites. Not only has patent filarial infection been shown to modulate T cell responses, but it has also been shown to result in profound monocyte dysfunction that can be reversed by effective treatment with anti-filarial chemotherapy. Subsequently we have shown that LF induces an immunoregulatory population of monocytes that appear to be human parallels of alternatively activated macrophages. In addition, LF is associated with an expansion of CCR1 low circulating myeloid DCs 6 suggesting the myeloid populations may be of great import in the modulation of filarial-specific T cell function. Expansion of a non-classical (CD16hi) monocyte subset has very recently been identified in LF. Moreover, we have shown that in human monocytes populations the major 2 subsets (CD14+CD16- and CD14+CD16lo) have a clearly differential response to microfilariae (Mf) , the latter preferentially producing the regulatory cytokine IL-10. Using human monocyte derived dendritic cells (DC), we have explored the mechanisms by which parasite products alter the function of human APCs. Having previously shown that mf-derived soluble products interfere directly with TLR3 and TLR4 signaling pathways, we were next able to demonstrate that parasite products profoundly modulated the DCs ability to produce IL-12, CCL10 and CCL11 8 in response to antigen or TLR agonists. By using approaches to silence important transcription factors, we were able to demonstrate the parasite-induced diminution of IRF-1 in DCs dramatically alters IL-12 production thereby preventing INF-gamma (and other Th1-associated) responses in LF. Moreover, using global protein expression profiling comparisons between Mf-exposed or unexposed human mDCs, mf significantly downregulated the mammalian target of rapamycin (mTOR) and the eukaryotic initiation factor (eIF) 2, eIF4 and p70S6K. Interestingly, live mf produce and secrete homologues of human FKBP1 (cyclophilin) a negative regulator of mTOR signaling. The effect of chronicity of infection per se on the T cell response to filarial infection has been studied by an exhaustive comparison of gene expression between cells purified from those with lifelong filarial infection and those expatriates with relatively acute infection. Using gene expression profiling, we were able to show that longstanding filarial infection (based on global gene expression ex vivo) showed a markedly down-regulated pattern of gene expression most notably in CD8+ cells but also in CD4+cells. Because chronic filarial (and other helminth) infections may alter immune reactivity to other (non-parasite) antigens and because these alterations may have profound implications for the clinical outcome of these non-filarial infections, collaborative studies in India, Mali, and Ecuador have shown that the presence of active filarial infection and/or chronic intestinal helminth infection very clearly blunts the Type 1 (and Th17) response to non-filarial antigens in the context of co-infection. Over the past four years, we have focused on the influence of pre-existing helminth infections on Mycobacterium tuberculosis (Mtb), malaria, and HIV. As a first step in examining the interaction between Mtb and filarial infection and based on the epidemiology of filarial/Mtb coinfection, an in vitro system of co infection was established and used to demonstrate that pre-exposure of human DCs and macrophages to live filarial parasites induces immunoregulatory phenotypes that alters mycobacterial entry and replication (Chatterjee, unpublished). Concurrently, we developed an in vivo model in which we rendered mice microfilaremic and then infected them (by aerosol) with virulent Mtb. Our data very clearly suggests that microfilaremia induces alternative activation of macrophages in the lung but fails to alter the growth or clearance of Mtb; microarray data from the lungs of these mice suggest a generalized modulation of interferon-gamma and IFN-gamma; regulated pathways (Chatterjee, Talaat, unpublished). Human studies ex vivo focusing more on latent tuberculosis in filarial-infected and -uninfected individuals have demonstrated that filarial infection (and gastrointestinal nematodes as well) modulates mycobacterial-speidvid responses. We have now extended these studies by developing multiparameter flow cytometric approaches to identify Mtb-specific stem cell memory (SCM) CD4+ and CD8+ T cells and are exploring the relationship between chronic filarial antigen exposure and the alteration in Mtb-specific memory

在患有专利淋巴丝虫病个体中看到的免疫反应的标志是对寄生虫抗原的反应，无法增殖或产生与1型反应（IL 2和IFN-GAMMA;）相关的细胞因子。这种寄生虫特异性的不反应在很大程度上由TGF-beta介导IL 10。和CTLA-4扮演较小的监管角色。 IL-10超家族（IL-19和IL-24）的其他成员现已显示在专利LF中被上调，这是由IL-10本身驱动的过程。 LF中寄生虫特异性反应调节的基础机制仍然是一个主要的研究问题。使用多参数流式细胞术，该抗原特异性调制显示与产生自适应Treg的IL-10的扩展以及寄生虫特异性中心和效应的记忆T细胞种群的发展有关。我们已经证明了这些Treg会影响效应T细胞和抗原呈递细胞（APC），但通过效应T细胞间接改变APC功能（Metenou等，提交）。最近，我们已经在丝状感染的背景下使用NTREG的蛋白质表达分析（通过LC/MS/MS）确定，由NTREG分泌的另外2个分子（WISP3和PSG-1）抑制T细胞增殖和IFN-GAMMA-GAMMA的nTREG分泌，并以接触方式独立于接触方式。我们还表明，这些丝状感染中的Ntregs高度异质和激活，但被编程为短暂的。这可能是其数量增加的基础，这是丝状寄生虫感染的特征。不仅证明了专利的丝状感染可以调节T细胞反应，而且还显示出严重的单核细胞功能障碍，可以通过有效治疗抗抗性化学疗法来逆转。随后，我们表明LF诱导了单核细胞的免疫调节群，这些人群似乎是人类的巨噬细胞的人类相似之处。此外，LF与CCR1低循环髓样DCS 6的扩展有关，表明髓样群在调制丝状特异性T细胞功能时可能非常重要。最近在LF中发现了非古典（CD16HI）单核细胞子集的扩展。此外，我们已经表明，在人类单核细胞种群中，主要的2个子集（CD14+CD16和CD14+CD16LO）对微毛利亚（MF）具有明显的差异反应，后者优先生产调节性细胞因子IL-10。使用人类单核细胞衍生的树突状细胞（DC），我们探索了寄生产物改变人APC功能的机制。先前表明MF衍生的可溶性产物直接干扰TLR3和TLR4信号通路，因此我们接下来能够证明寄生虫产物对DCS产生IL-12，CCL10和CCL11 8的能力深刻调节，以响应抗原或TLR激动剂。通过使用沉默重要转录因子的方法，我们能够证明DCS中寄生虫引起的IRF-1降低显着改变IL-12的产生，从而防止LF中INF-GAMMA（和其他TH1相关）响应。此外，使用MF暴露或未暴露的人MDC之间的全球蛋白表达分析比较，MF显着下调了雷帕霉素的哺乳动物靶标（MTOR）（MTOR）和真核生物起始因子（EIF）2，EIF4和P70S6K。有趣的是，人类FKBP1（环磷脂）的活MF产物和分泌同源物是MTOR信号传导的负调节剂。通过详尽地比较了从终生丝状感染的细胞和相对急性感染的外籍者之间纯化的细胞之间的基因表达，研究了感染本身对T细胞对丝状感染的反应的影响。使用基因表达分析，我们能够证明长期存在的丝状感染（基于全局基因表达）显示出明显下调基因表达的模式，最著名的是在CD8+细胞中，而在CD4+细胞中也是如此。因为慢性丝状（和其他蠕虫感染）可能会改变对其他（非寄生虫）抗原的免疫反应性，并且因为这些变化可能对这些非狂热感染的临床结果产生深远的影响，印度，马里和eCuador的协作研究表明，有活跃的丝状感染和/或/或/或/均具有炎症感染的影响，并具有1次均为/或/或/或/或均匀的直肠感染。在共同感染的背景下非抗原抗原。在过去的四年中，我们专注于先前存在的蠕虫感染对结核分枝杆菌（MTB），疟疾和HIV的影响。 As a first step in examining the interaction between Mtb and filarial infection and based on the epidemiology of filarial/Mtb coinfection, an in vitro system of co infection was established and used to demonstrate that pre-exposure of human DCs and macrophages to live filarial parasites induces immunoregulatory phenotypes that alters mycobacterial entry and replication (Chatterjee,未出版）。同时，我们开发了一个体内模型，在该模型中，我们使小鼠微毛皮流血，然后用毒气MTB感染（气溶胶）。我们的数据非常清楚地表明，微丝虫病会诱导肺中巨噬细胞的替代激活，但未能改变MTB的生长或清除。这些小鼠肺的微阵列数据表明，干扰素伽马和IFN-gamma的广义调节。规范的途径（Chatterjee，Talaat，未出版）。人类研究在丝状感染和未感染的个体中更多地关注潜在的结核病，这表明丝状感染（以及胃肠道线虫也）调节了分枝杆菌 - 细菌 - 细菌 - 细胞杆菌反应。现在，我们通过开发多参数流式细胞仪方法来扩展这些研究，以鉴定MTB特异性干细胞记忆（SCM）CD4+和CD8+ T细胞，并正在探索慢性丝质抗原暴露与MTB特异性内存改变之间的关系