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: 10272013
• 负责人: Thomas Nutman
• 金额: $ 110.69万
• 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/10272013
• 关键词: Adult; Affect; Allergens; Allergic; Allergic Disease; Amino Acid Sequence; Antigens; Ascaris; Automobile Driving; Baculoviruses; Binding; Binding Proteins; Biological Assay; Brugia malayi; CCL1 gene; CCL11 gene; CCL2 gene; CCL4 gene; CXCL10 gene; CXCL11 gene; CXCL9 gene; Clinical; Data; Dendritic Cells; Development; Disease; EIF4EBP1 gene; Electron Microscopy; Extrinsic asthma; FRAP1 gene; Family member; Filarial Elephantiases; Flow Cytometry; Functional disorder; Genetic; Goals; Helminths; Human; Immobilization; Immune; Immune response; Immunity; Immunologics; Individual; Infection; Inflammation; Inflammation Mediators; Interleukin-10; Interleukin-13; Interleukin-2; Interleukin-4; Interleukin-5; Interleukin-9; Larva; Link; Liquid Chromatography; Liver; Loiasis; Lung; Lymphatic; Malaria; Mediating; Methods; MicroRNAs; Microfilaria; Morphology; Mus; Mycobacterium tuberculosis; Nature; Non-Insulin-Dependent Diabetes Mellitus; Nucleic Acids; Onchocerciasis; Outcome; PD-1/PD-L1; PDL1 pathway; Parasites; Parasitic infection; Pathogenesis; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Phase; Phosphorylation; Play; Production; Protein-Serine-Threonine Kinases; Proteins; Proteomics; Pyroglyphidae; Reading; Regulation; Role; STAT3 gene; Signal Transduction; Sirolimus; Solid; Strongyloides; Strongyloidiasis; Structure; T-Lymphocyte; Tissues; Tuberculosis; Vesicle; Western Blotting; Work; cell growth; chemokine; co-infection; cytokine; dimer; eosinophil; exosome; exposed human population; genetic regulatory protein; helminth infection; immunoregulation; interleukin-13 receptor; interleukin-19; mTOR Inhibitor; mTOR inhibition; member; monocyte; mouse model; programs; response; tandem mass spectrometry; transcriptome sequencing; transcriptomics

The overriding goal of this project is to provide an understanding of how the immunologic responses to filarial and related parasites are controlled. The major aspects of this project involve the parasite-specific responses in lymphatic filariasis (LF), loiasis, onchocerciasis and most recently strongyloidiasis in terms of regulation, pathogenesis protective immunity, and the genetic underpinnings of these host responses. The major objectives are to identify the mechanisms by which the modulation/regulation of immune responsiveness works in filarial and related parasitic infections; 2) to identify factors involved in the pathogenesis of disease in filarial infections; 3) to identify the role of host and parasite factors underlying the differential responsiveness to parasite antigens and the subsequent clinical and immunologic outcome; and 4) to understand the immunologic correlates of immunity in human filarial infection. IL-10 has been shown to be the primary immunoregulatory cytokine driving the modulation of the host response in many filarial infections. To determine whether parasite-derived molecules could modulate the host IL-10/IL-10R pathways, we performed solid-phase immobilization of human IL-10Ra followed by binding assays with a Brugia malayi (Bm) adult antigen extracts. Bm proteins that bound to human IL-10Ra were eluted and analyzed by liquid chromatography-tandem mass spectrometry. Our proteomic analysis identified 5 Bm molecules that bound human IL-10Ra. Using a structural alignment program, we identified a 164 amino acid sequence from Bm5539 (the top hit) that shared high structural homology with the human IL-10 functional dimer. Using a baculovirus-expressed truncated form of Bm5539, we examined its ability to signal through the human IL-10R, reading out phosphorylation of STAT3 (pSTAT3) in human monocytes by flow cytometry, and showed that Bm5539 appears to be an IL-10 antagonist. Furthermore, as IL-10 belongs to a superfamily of cytokines and 2 of the IL-10 superfamily members IL-24 and IL-19 are increased in active (antigen positive) W. bancrofti infections and to understand the role played by microfilaria (mf) in their induction, peripheral blood mononuclear cells from patients with filarial infections were stimulated with or without filarial antigen they were shown to produce significant levels of IL-10, IL-13, IL-5. IL-4, IL-9 IL-2, and IL-27. When comparing mf positive (mf+) and mf negative (mf-) patients, there were no significant differences in most of the cytokines; in marked contrast, mf+ individuals had significantly increased filarial antigen-driven IL-24 and IL-19 (compared to mf- subjects. These data provide an important link between IL-10 and its related family members IL-19 and IL-24 in the modulation of the immune response in human filarial infections that appears to be driven by microfilariae. Although IL-10 plays a major role in the altering the response to both both filarial antigen and bystander antigens/diseases (e.g. allergic diseases, Mtb, malaria), we developed methods to understand more fully this issue. Using a murine model of house dust mite (HDM)-induced allergic asthma inflammation followed by Ascaris infection, we demonstrated that pre-existing allergen sensitization markedly limits Ascaris larval development and reduces up to 70% the parasite burden in the lung. This reduction in parasite burden is driven by an eosinophil-dependent pulmonary type-2-immune response. To explore further the role played by IL-13/IL-13R signaling in mediating this eosinophil-dependent phenomenon, we show that in HDM-sensitized-IL-13Ra1 deficient mice, there was a significant reduction in tissue eosinophil number (a reduction similar to that found in HDM-sensitized eosinophil-deficient mice) that, as a consequence. failed to limit parasite development or numbers. RNA-seq analyses of Ascaris larvae isolated from the lungs of allergen pre-sensitized mice compared to larvae from non-allergic mice showed a transcriptomic-signature of the L3-liver stage. Our data suggest that HDM-induced allergic sensitization drives a response that mimics a primary Ascaris infection, such that lung-specific IL-13Ra1 signaling driven by allergen sensitization mediates eosinophil-dependent helminth larval killing in the tissue occurs. In related human studies, in which we examined how helminth infections are known to modulate T cell and cytokine responses in latent Mycobacterium tuberculosis infection (LTBI) and because chemokines play a vital role in the protective immune responses in LTBI, we demonstrated that in LTBI with concomitant Strongyloides infection has a myriad number of affects includingL 1) causing a marked decrease in the production of the CCL1, CCL2, CCL4, CCL11, CXCL9, CXCL10, and CXCL11 chemokines; 2) altering the nature of monocyte activation; 3) modulating the pro-inflammatory mediators of Type 2 diabetes; 4) altering the PD1/PDL1 pathway in the context of co-infection. We have previously shown that the microfilarial (mf) stage of Brugia malayi can inhibit the mammalian target of rapamycin (mTOR; a conserved serine/threonine kinase critical for immune regulation and cellular growth) in human dendritic cells (DC) and we have proposed that this mTOR inhibition is associated with the DC dysfunction seen in filarial infections. Because exosome-like vesicles (ELV) contain many proteins and nucleic acids such as microRNAs (miRNAs) that affect a variety of cellular pathways, we hypothesized that ELV secreted from mf are enriched in miRNAs that target and downregulate the mTOR pathway in human DC. Thus, ELV, purified from mf of Brugia malayi and confirmed by morphology and size through electron microscopy and Nanosight analysis, were shown to be enriched for miR100, miR71, miR34, miR7, and let-7. After confirming their presence in ELVs using qPCR for these miRNA targets, target predictions suggested that mir100 and let-7 targeted mTOR and its downstream regulatory protein 4-EBP1 respectively. We then exposed human DC and monocytes to these ELVs. After confirming the internalization of mf-derived ELV, we were able to demonstrate through western blotting and flow cytometry that mf ELV downregulate the phosphorylation of 4E-BP1 and mTOR to the same degree that rapamycin (a known mTOR inhibitor) does. Our data collectively suggest that mf ELV are enriched in miRNAs that regulate mTOR.

本项目的首要目标是提供对丝虫和相关寄生虫的免疫反应是如何控制的理解。该项目的主要方面涉及淋巴丝虫病（LF）、loloasis、盘尾丝虫病和最近的圆线虫病的寄生虫特异性反应，涉及这些宿主反应的调节、发病机制、保护性免疫和遗传基础。主要目标是确定在丝虫病和相关寄生虫感染中调节/调节免疫反应的机制；2)确定丝虫病感染中涉及疾病发病机制的因素；3)确定宿主和寄生虫因素对寄生虫抗原的差异反应以及随后的临床和免疫结果的作用；4)了解人丝虫病感染免疫的免疫学相关因素。