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

• 批准号: 8745274
• 负责人: Thomas Nutman
• 金额: $ 88.93万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 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/8745274
• 关键词: 70-kDa Ribosomal Protein S6 Kinases; Acute; Aerosols; Agonist; Antigen-Presenting Cells; Antigens; Bystander Effect; CCL11 gene; CCR1 gene; CD14 gene; CD3 Antigens; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Cell physiology; Cells; Chronic; Clinical; Cook Islands; Cryopreserved Cell; 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; IL2RB gene; IL7R gene; IRF1 gene; Immune; Immune response; Immunity; Immunologics; In Vitro; India; Individual; Infection; Infection prevention; Inflammatory; Interferon Type II; Interferons; Interleukin-10; Interleukin-12; Intestines; Legal patent; Life; Loiasis; Lung; Lymphatic; Macrophage Activation; Malaria; Mali; Mediating; Memory; Microfilaria; Molecular Profiling; Mus; Mycobacterium tuberculosis; Mycobacterium tuberculosis antigens; Myelogenous; Onchocerciasis; Outcome; Parasite Control; Parasites; Parasitic infection; Pathway interactions; Patients; Phenotype; Play; Population; Process; Production; Proliferating; Regulation; Regulatory T-Lymphocyte; Research; Role; Seasons; Signal Pathway; Signal Transduction; Site; Stem cells; System; T cell response; T memory cell; T-Cell Proliferation; T-Lymphocyte; TLR3 gene; TLR4 gene; Tuberculosis; Viral Tumor Antigens; Virulent; Virus Diseases; WISP3 gene; Work; anergy; base; chemotherapy; cytokine; effective therapy; exposed human population; filaria; immunoregulation; in vivo Model; interleukin-19; liquid chromatography mass spectrometry; mTOR protein; macrophage; member; monocyte; mycobacterial; prevent; protein expression; response; transcription factor; transmission process

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 γ) in response to parasite antigen. This parasite specific anergy is mediated, in large part, by IL 10 with TGF β 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 4. 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−γ production by effector T cells in a contact independent manner. 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 (review here) 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) 7, 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-γ (and other Th1-associated) responses in LF 9. 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/ In a study that has taken advantage of cells cryopreserved from well characterized individuals from the Cook Islands where W. bancrofti is endemic, we have utilized multiparameter flow to demonstrate not only a diminished number of central memory cells (CD4+CCR7+CD127+CD27+CD45RA-) in chronically infected patients but also an increase number of revertant memory cells (CD3+ CD45RA+CCR7-CD127+CD122+) suggesting that chronicity in helminth infection (like in chronic viral infection) may predispose to memory cell dysfunction. 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-γ regulated pathways (Chatterjee, Talaat, unpublished). Human studies ex vivo focusing more on latent tuberculosis in filarial-infected and -uninfected individuals have demonstrated that filarial infections: 1) modulate TLR expression and function in response to Mtb antigens 12; 2) markedly alter the Mtb-specific Th1/Th17 responses so important in maintaining latency through the concerted actions of CTLA-4 and PD-1 2; and 3) induce a population of Tregs that modulate Th17 responses in patients with latent Mtb infection 13. 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 In studies in Mali that have combined clinical and immunological aspects of malaria and filarial co-infection, our immunologically-focused studies have explored the bystander effects of pre-existing filarial infections on the response to malarial antigens longitudinally (before, during, and following the malarial transmission season). In two studies performed in different sites two years apart, we have shown quite conclusively that the presence of filarial infection modulates the innate (TLR-mediated) response to malarial antigens; more importantly, however, we have shown that filarial infection modulates malaria-specific Type 1 cytokine responses in an IL-10 dependent manner in a filaria/malaria co-infected population. We have identified the cell populations regulating the malaria-specific pro-inflammatory cytokine pathways in the co-filarial/malaria co-infected populations, and we have also demonstrated that filarial infections prevent almost completely the induction of malaria-specific polyfunctional T cells. Furthermore, we have demonstrated the pivotal role played by IRF-1 in the modulation of the malaria-specific Th1/Th17 responses.

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（IL 2和IFNγ）的增殖或产生与1型反应（IL 2和IFNγ）相关的细胞因子。在很大程度上，IL 10与TGFβ和CTLA-4扮演较小的调节作用介导了这种寄生虫特异性的反应。 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。 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 4. 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).最近，我们使用了NTREG的蛋白质表达分析（通过LC/MS/MS）在丝状感染的背景下，由NTREG抑制T细胞增殖和IFN-γ产生的NTREG分泌的另外2个分子（WISP3和PSG-1）以独立的方式抑制了IFN-γ。 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（此处综述）可以通过有效的抗饮食化学疗法进行有效治疗来逆转。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2个子集（CD14+CD16和CD14+CD16LO）对微毛利亚（MF）7具有明显的差异响应，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。 通过使用静默的重要转录因子的方法，我们能够证明DC中寄生虫引起的IRF-1的减少显着改变IL-12的产生，从而防止LF 9中的INF-γ（和其他Th1相关的）响应。更多地，使用MF暴露于MF的人类MF，MF的全球蛋白质表达靶向，MF的全球蛋白质表达靶向，MF在MF中，MF的靶标的靶向，MF均具有强大的MF，MF在MF中，MF的MF在MF中的强大启示。 （MTOR）和真核开始因子（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。使用基因表达谱分析，我们能够证明长期丝状感染（基于全球基因表达离体）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/ 在一项利用W. bancrofti是地方性库克岛的良好表征个体中的细胞中，我们利用多参数流的良好个体，不仅证明了中枢记忆细胞数量减少（CD4+CCR7+CCR7+CD127+CD27+CD27+CD45RA-），还增加了年代感染的患者，还增加了reververnational Memory Memborn Memory（CD+3+3+3） CD45RA+CCR7-CD127+CD122+）表明，蠕虫感染（如慢性病毒感染）可能会易于记忆细胞功能障碍。 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。来自这些小鼠肺的微阵列数据表明，干扰素伽马和IFN-γ调控途径的广义调节（Chatterjee，Talaat，未发表）。 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，证明了丝状感染：1）根据MTB抗原12调节TLR表达和功能。 2）明显改变了MTB特异性的TH1/TH17响应，对于通过CTLA-4和PD-1 2的一致作用保持潜伏期至关重要； 3）诱导潜在MTB感染患者调节Th17反应的Treg人群13。我们现在通过开发多参数计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，并正在探索慢性丝状抗原曝光与MTB的慢性抗原曝光与MTB替代品之间的关系 在将疟疾的临床和免疫学方面结合在一起的马里研究中，我们以免疫学为中心的研究探索了先前存在的丝状感染对疟疾抗原纵向反应的旁观者的影响（在疟疾传输季节之前和之后）。 在两年相隔两年的不同地点进行的两项研究中，我们已经很确切地表明，丝状感染的存在调节了先天性（TLR介导的）对疟疾抗原的反应。但是，更重要的是，我们已经表明，丝状感染以IL-10依赖性方式调节疟疾特异性1型细胞因子反应，以丝状/疟疾共感染的种群调节。 我们已经确定了调节共鼠/疟疾共感染群体中疟疾特异性促炎细胞因子途径的细胞群体，我们还证明，丝状感染几乎完全防止了疟疾特异性多功能T细胞的诱导。 此外，我们已经证明了IRF-1在调节疟疾特异性TH1/TH17反应中所起的关键作用。