ABSTRACT
Interleukin-23 (IL-23) and IL-17 are cytokines currently being targeted in clinical trials. Although inhibition of both of these cytokines is effective for treating psoriasis, IL-12 and IL-23 p40 inhibition attenuates Crohn's disease, whereas IL-17A or IL-17 receptor A (IL-17RA) inhibition exacerbates Crohn's disease. This dichotomy between IL-23 and IL-17 was effectively modeled in the multidrug resistance-1a-ablated (Abcb1a(-/-)) mouse model of colitis. IL-23 inhibition attenuated disease by decreasing colonic inflammation while enhancing regulatory T (Treg) cell accumulation. Exacerbation of colitis by IL-17A or IL-17RA inhibition was associated with severe weakening of the intestinal epithelial barrier, culminating in increased colonic inflammation and accelerated mortality. These data show that IL-17A acts on intestinal epithelium to promote barrier function and provide insight into mechanisms underlying exacerbation of Crohn's disease when IL-17A or IL-17RA is inhibited.
Subject(s)
Colitis/immunology , Interleukin-17/physiology , Interleukin-23/physiology , Receptors, Interleukin-17/physiology , ATP Binding Cassette Transporter, Subfamily B/deficiency , Animals , Colitis/drug therapy , Colitis/etiology , Colitis/microbiology , Disease Models, Animal , Disease Progression , Epithelium/physiopathology , Female , Forkhead Transcription Factors/analysis , Gene Expression Regulation/immunology , Helicobacter Infections/complications , Helicobacter Infections/immunology , Immunization, Passive , Immunoglobulin G/therapeutic use , Interleukin-12 Subunit p40/antagonists & inhibitors , Interleukin-17/immunology , Interleukin-23/immunology , Interleukin-23 Subunit p19/antagonists & inhibitors , Interleukin-23 Subunit p19/immunology , Intestinal Mucosa/physiopathology , Mice , Mice, Knockout , Permeability , Receptors, Interleukin-17/antagonists & inhibitors , Receptors, Interleukin-17/immunology , T-Lymphocyte Subsets/immunology , T-Lymphocytes, Regulatory/immunology , TranscriptomeABSTRACT
Severe asthma is associated with the production of interleukin 17A (IL-17A). The exact role of IL-17A in severe asthma and the factors that drive its production are unknown. Here we demonstrate that IL-17A mediated severe airway hyperresponsiveness (AHR) in susceptible strains of mice by enhancing IL-13-driven responses. Mechanistically, we demonstrate that IL-17A and AHR were regulated by allergen-driven production of anaphylatoxins, as mouse strains deficient in complement factor 5 (C5) or the complement receptor C5aR mounted robust IL-17A responses, whereas mice deficient in C3aR had fewer IL-17-producing helper T cells (T(H)17 cells) and less AHR after allergen challenge. The opposing effects of C3a and C5a were mediated through their reciprocal regulation of IL-23 production. These data demonstrate a critical role for complement-mediated regulation of the IL-23-T(H)17 axis in severe asthma.
Subject(s)
Asthma/immunology , Complement C3a/immunology , Complement C5a/immunology , Interleukin-17/biosynthesis , Interleukin-23/immunology , Th2 Cells/immunology , Allergens/adverse effects , Anaphylatoxins/biosynthesis , Animals , Asthma/genetics , Complement Activation , Complement C3a/genetics , Complement C5a/genetics , Cytokines/biosynthesis , Genetic Predisposition to Disease , Interleukin-13/biosynthesis , Interleukin-17/genetics , Male , Mice , Mice, Inbred AKR , Mice, Inbred BALB C , Mice, Inbred C3H , Mice, Inbred C57BL , Mice, Inbred DBA , Mice, Knockout , Pyroglyphidae/immunology , Receptor, Anaphylatoxin C5a/genetics , Th2 Cells/metabolismABSTRACT
BACKGROUND: Allergic sensitization to fungi has been associated with asthma severity. As a result, it has been largely assumed that the contribution of fungi to allergic disease is mediated through their potent antigenicity. OBJECTIVE: We sought to determine the mechanism by which fungi affect asthma development and severity. METHODS: We integrated epidemiologic and experimental asthma models to explore the effect of fungal exposure on asthma development and severity. RESULTS: We report that fungal exposure enhances allergen-driven TH2 responses, promoting severe allergic asthma. This effect is independent of fungal sensitization and can be reconstituted with ß-glucan and abrogated by neutralization of IL-17A. Furthermore, this severe asthma is resistant to steroids and characterized by mixed TH2 and TH17 responses, including IL-13+IL-17+CD4+ double-producing effector T cells. Steroid resistance is dependent on fungus-induced TH17 responses because steroid sensitivity was restored in IL-17rc-/- mice. Similarly, in children with asthma, fungal exposure was associated with increased serum IL-17A levels and asthma severity. CONCLUSION: Our data demonstrate that fungi are potent immunomodulators and have powerful effects on asthma independent of their potential to act as antigens. Furthermore, our results provide a strong rationale for combination treatment strategies targeting IL-17A for this subgroup of fungus-exposed patients with difficult-to-treat asthma.
Subject(s)
Allergens/immunology , Asthma/immunology , Fungi/immunology , Th17 Cells/immunology , Th2 Cells/immunology , beta-Glucans/immunology , Air Pollutants/immunology , Animals , Anti-Inflammatory Agents/therapeutic use , Antigens, Dermatophagoides/immunology , Asthma/drug therapy , Asthma/epidemiology , Asthma/pathology , Child , Child, Preschool , Dexamethasone/therapeutic use , Drug Resistance/immunology , Environmental Exposure , Female , Glucocorticoids/therapeutic use , Humans , Infant , Interleukin-17/blood , Interleukin-17/immunology , Lectins, C-Type/genetics , Lung/drug effects , Lung/immunology , Lung/pathology , Male , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , Prevalence , Receptors, Interleukin/geneticsABSTRACT
IL-17 cytokines are expressed by a variety of cells and mediate host defence against extracellular pathogens. IL-17 is upregulated at sites of inflammation and can synergize with other cytokines, such as TNF-α, to amplify the inflammatory response. Activation of these signalling pathways has been hypothesized to contribute to the underlying pathogenesis of several inflammatory diseases, including psoriasis, RA, PsA and asthma. Thus the IL-17 signalling pathway is an attractive target for the development of therapeutic agents to modulate aberrant inflammatory responses. This review of the clinical development of therapeutic agents that target IL-17 signalling pathways in inflammatory diseases focuses on brodalumab, a human anti-IL-17 receptor A mAb. The cumulative findings of early clinical studies with anti-IL-17 agents, including brodalumab, secukinumab and ixekizumab, provide strong evidence for the role of IL-17 signalling in the pathophysiology of certain inflammatory diseases and support the potential use of these agents in treating these diseases.
Subject(s)
Antibodies, Monoclonal/therapeutic use , Arthritis, Psoriatic/drug therapy , Arthritis, Rheumatoid/drug therapy , Asthma/drug therapy , Crohn Disease/drug therapy , Receptors, Interleukin-17/antagonists & inhibitors , Antibodies, Monoclonal, Humanized/therapeutic use , Arthritis, Psoriatic/immunology , Arthritis, Rheumatoid/immunology , Asthma/immunology , Crohn Disease/immunology , Humans , Molecular Targeted Therapy , Psoriasis/drug therapy , Psoriasis/immunology , Receptors, Interleukin-17/immunologyABSTRACT
CD4(+) T helper 2 (T(H)2) cells secrete interleukin (IL)4, IL5 and IL13, and are required for immunity to gastrointestinal helminth infections. However, T(H)2 cells also promote chronic inflammation associated with asthma and allergic disorders. The non-haematopoietic-cell-derived cytokines thymic stromal lymphopoietin, IL33 and IL25 (also known as IL17E) have been implicated in inducing T(H)2 cell-dependent inflammation at mucosal sites, but how these cytokines influence innate immune responses remains poorly defined. Here we show that IL25, a member of the IL17 cytokine family, promotes the accumulation of a lineage-negative (Lin(-)) multipotent progenitor (MPP) cell population in the gut-associated lymphoid tissue that promotes T(H)2 cytokine responses. The IL25-elicited cell population, termed MPP(type2) cells, was defined by the expression of Sca-1 (also known as Ly6a) and intermediate expression of c-Kit (c-Kit(int)), and exhibited multipotent capacity, giving rise to cells of monocyte/macrophage and granulocyte lineages both in vitro and in vivo. Progeny of MPP(type2) cells were competent antigen presenting cells, and adoptive transfer of MPP(type2) cells could promote T(H)2 cytokine responses and confer protective immunity to helminth infection in normally susceptible Il25(-/-) mice. The ability of IL25 to induce the emergence of an MPP(type2) cell population identifies a link between the IL17 cytokine family and extramedullary haematopoiesis, and suggests a previously unrecognized innate immune pathway that promotes T(H)2 cytokine responses at mucosal sites.
Subject(s)
Cell Differentiation , Interleukins/immunology , Multipotent Stem Cells/cytology , Multipotent Stem Cells/immunology , Th2 Cells/immunology , Th2 Cells/metabolism , Animals , Antigens, Ly/metabolism , Cell Lineage , Granulocytes/cytology , Granulocytes/immunology , Granulocytes/metabolism , Immunity, Innate/immunology , Immunity, Mucosal/immunology , Interleukins/biosynthesis , Interleukins/metabolism , Lymphoid Tissue/cytology , Lymphoid Tissue/immunology , Macrophages/cytology , Macrophages/immunology , Macrophages/metabolism , Membrane Proteins/metabolism , Mice , Mice, Inbred Strains , Nippostrongylus/immunology , Proto-Oncogene Proteins c-kit/metabolism , Strongylida Infections/immunology , Th2 Cells/cytology , Trichuriasis/immunology , Trichuris/immunologyABSTRACT
Asthma is a heterogeneous disease characterized by airway inflammation and hyperreactivity. IL-17 receptor A (IL-17RA) is a shared receptor subunit required for activity of IL-17 family cytokines, including IL-17A and IL-25. IL-17A and IL-25 induce different proinflammatory responses, and concentrations are elevated in subjects with asthma. However, the individual contributions of IL-17A and IL-25 to disease pathogenesis are unclear. We explored proinflammatory activities of the IL-17 pathway in models of pulmonary inflammation and assessed its effects on contractility of human bronchial airway smooth muscle. In two mouse models, IL-17RA, IL-17RB, or IL-25 blockade reduced airway inflammation and airway hyperreactivity. Individually, IL-17A and IL-25 enhanced contractility of human bronchial smooth muscle induced by methacholine or carbachol. IL-17A had more pronounced effects on methacholine-induced contractility in bronchial rings from donors with asthma compared with donors without asthma. Blocking the IL-17 pathway via IL-17RA may be a useful therapy for some patients with asthma by reducing pulmonary inflammation and airway hyperreactivity.
Subject(s)
Asthma/metabolism , Receptors, Interleukin-17/physiology , Animals , Asthma/immunology , Bronchi/immunology , Bronchi/pathology , Cells, Cultured , Gene Expression , Humans , Interleukin-17/physiology , Interleukins/physiology , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , Muscle Contraction , Myocytes, Smooth Muscle/immunology , Myocytes, Smooth Muscle/metabolism , Signal TransductionABSTRACT
Based on their structural similarity to previously described compound AMG 009, indole-phenyl acetic acids were proposed to be potent dual inhibitors of chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2 or DP2) and prostanoid D receptor (DP or DP1). This series was equipotent to AMG 009 in binding assays against both receptors but exhibited decreased serum shift. We discovered early in the optimization of these indole-phenylacetic acid compounds that they demonstrated CYP3A4 time-dependent inhibition (TDI). Hypothesizing that the source of TDI was the indole core we modified the 1,2,3-substitution to eventually afford a highly potent modulator of CRTH2 and DP which did not exhibit TDI.
Subject(s)
Cytochrome P-450 CYP3A/metabolism , Enzyme Inhibitors/pharmacology , Indoles/pharmacology , Phenylacetates/pharmacology , Receptors, Immunologic/antagonists & inhibitors , Receptors, Prostaglandin/antagonists & inhibitors , Dose-Response Relationship, Drug , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/chemistry , Humans , Indoles/chemistry , Molecular Structure , Phenylacetates/chemistry , Structure-Activity Relationship , Time FactorsABSTRACT
There is considerable evidence supporting a role for mold exposure in the pathogenesis and expression of childhood asthma. Aspergillus versicolor and Cladosporium cladosporioides are common molds that have been implicated in asthma. In a model of mold-induced asthma, mice were repeatedly exposed to either A. versicolor or C. cladosporioides spores. The two molds induced distinct phenotypes, and this effect was observed in both BALB/c and C57BL/6 strains. C. cladosporioides induced robust airway hyperresponsiveness (AHR), eosinophilia, and a predominately Th2 response, whereas A. versicolor induced a strong Th17 response and neutrophilic inflammation, but very mild AHR. Neutralization of IL-17A resulted in strong AHR and eosinophilic inflammation following A. versicolor exposure. In Dectin-1-deficient mice, A. versicolor exposure resulted in markedly attenuated IL-17A and robust AHR compared with wild-type mice. In contrast, C. cladosporioides induced AHR and eosinophilic inflammation independent of IL-17A and Dectin-1. A. versicolor, but not C. cladosporioides, spores had increased exposure of ß-glucans on their surface and were able to bind Dectin-1. Thus, the host response to C. cladosporioides was IL-17A- and Dectin-1-independent, whereas Dectin-1- and IL-17A-dependent pathways were protective against the development of asthma after exposure to A. versicolor.
Subject(s)
Anti-Asthmatic Agents/administration & dosage , Aspergillus/immunology , Asthma/immunology , Asthma/pathology , Cladosporium/immunology , Interleukin-17/administration & dosage , Lectins, C-Type/administration & dosage , beta-Glucans/administration & dosage , Animals , Anti-Asthmatic Agents/metabolism , Aspergillus/metabolism , Asthma/prevention & control , Bronchial Hyperreactivity/immunology , Bronchial Hyperreactivity/pathology , Bronchial Hyperreactivity/prevention & control , Cladosporium/metabolism , Eosinophils/immunology , Eosinophils/pathology , Immunophenotyping , Inflammation Mediators/administration & dosage , Lectins, C-Type/deficiency , Lectins, C-Type/metabolism , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , Neutrophils/immunology , Neutrophils/pathology , Spores, Fungal/immunology , Spores, Fungal/metabolism , Surface Properties , beta-Glucans/metabolismABSTRACT
BACKGROUND: IL-17A has been implicated in severe forms of asthma. However, the factors that promote IL-17A production during the pathogenesis of severe asthma remain undefined. Diesel exhaust particles (DEPs) are a major component of traffic-related air pollution and are implicated in asthma pathogenesis and exacerbation. OBJECTIVE: We sought to determine the mechanism by which DEP exposure affects asthma severity using human and mouse studies. METHODS: BALB/c mice were challenged with DEPs with or without house dust mite (HDM) extract. Airway inflammation and function, bronchoalveolar lavage fluid cytokine levels, and flow cytometry of lung T cells were assessed. The effect of DEP exposure on the frequency of asthma symptoms and serum cytokine levels was determined in children with allergic asthma. RESULTS: In mice exposure to DEPs alone did not induce asthma. DEP and HDM coexposure markedly enhanced airway hyperresponsiveness compared with HDM exposure alone and generated a mixed T(H)2 and T(H)17 response, including IL-13(+)IL-17A(+) double-producing T cells. IL-17A neutralization prevented DEP-induced exacerbation of airway hyperresponsiveness. Among 235 high DEP-exposed children with allergic asthma, 32.2% had more frequent asthma symptoms over a 12-month period compared with only 14.2% in the low DEP-exposed group (P = .002). Additionally, high DEP-exposed children with allergic asthma had nearly 6 times higher serum IL-17A levels compared with low DEP-exposed children. CONCLUSIONS: Expansion of T(H)17 cells contributes to DEP-mediated exacerbation of allergic asthma. Neutralization of IL-17A might be a useful potential therapeutic strategy to counteract the asthma-promoting effects of traffic-related air pollution, especially in highly exposed patients with severe allergic asthma.
Subject(s)
Asthma/etiology , Interleukin-17/biosynthesis , Vehicle Emissions , Adolescent , Allergens/immunology , Animals , Child , Child, Preschool , Cytokines/biosynthesis , Cytokines/blood , Disease Models, Animal , Female , Humans , Hyaluronan Receptors/metabolism , Inflammation/etiology , Inhalation Exposure/adverse effects , Interleukin-17/blood , L-Selectin/metabolism , Lung/immunology , Lung/metabolism , Male , Mice , Pyroglyphidae/immunology , Respiratory Hypersensitivity/etiology , Th17 Cells/immunology , Th17 Cells/metabolism , Th2 Cells/immunology , Th2 Cells/metabolism , Vehicle Emissions/toxicityABSTRACT
BACKGROUND: The D-prostanoid receptor and the chemoattractant receptor homologous molecule expressed on T(H)2 cells (CRTH2) are implicated in asthma pathogenesis. AMG 853 is a potent, selective, orally bioavailable, small-molecule dual antagonist of human D-prostanoid and CRTH2. OBJECTIVE: We sought to determine the efficacy and safety of AMG 853 compared with placebo in patients with inadequately controlled asthma. METHODS: Adults with moderate-to-severe asthma were randomized to placebo; 5, 25, or 100 mg of oral AMG 853 twice daily; or 200 mg of AMG 853 once daily for 12 weeks. All patients continued their inhaled corticosteroids. Long-acting ß-agonists were not allowed during the treatment period. Allowed concomitant medications included short-acting ß-agonists and a systemic corticosteroid burst for asthma exacerbation. The primary end point was change in total Asthma Control Questionnaire score from baseline to week 12. Secondary and exploratory end points included FEV(1), symptom scores, rescue short-acting ß-agonist use, and exacerbations. RESULTS: Among treated patients, no effect over placebo (n = 79) was observed in mean changes in Asthma Control Questionnaire scores at 12 weeks (placebo, -0.492; range for AMG 853 groups [n = 317], -0.444 to -0.555). No significant differences between the active and placebo groups were observed for secondary end points. The most commonly reported adverse events were asthma, upper respiratory tract infection, and headache; 9 patients experienced serious adverse events, all of which were deemed unrelated to study treatment by the investigator. CONCLUSION: AMG 853 as an add-on to inhaled corticosteroid therapy demonstrated no associated risks but was not effective at improving asthma symptoms or lung function in patients with inadequately controlled moderate-to-severe asthma.
Subject(s)
Anti-Asthmatic Agents/therapeutic use , Asthma/drug therapy , Phenylacetates/therapeutic use , Receptors, Immunologic/antagonists & inhibitors , Receptors, Prostaglandin/antagonists & inhibitors , Sulfonamides/therapeutic use , Adrenal Cortex Hormones/pharmacology , Adult , Anti-Asthmatic Agents/adverse effects , Asthma/physiopathology , Dose-Response Relationship, Drug , Double-Blind Method , Drug Therapy, Combination , Female , Humans , Male , Middle Aged , Phenylacetates/adverse effects , Respiratory Function Tests , Sulfonamides/adverse effectsABSTRACT
In the present studies local neutralization of allergen-induced stem cell factor (SCF) leads to decreased production of Th2 cytokines, a reduction in inflammation, allergen-specific serum IgE/IgG1, and attenuation of severe asthma-like responses. The local blockade of pulmonary SCF also resulted in a significant reduction of IL-17E (IL-25). Sorted cell populations from the lung indicated that IL-25 was produced from c-kit(+) cells, whereas Th2 cytokine production was primarily from c-kit(-) cell populations. SCF stimulated c-kit(+) eosinophils produced IL-25, whereas bone marrow-derived mast cells did not. Using 4get mice that contain a IL-4-IRES-eGFP that when transcribed coexpress GFP and IL-4, our studies identified cells that comprised a CD11b(+), GR1(+), Ly6C(+/-), c-kit(-), CD4(-), CD11c(-), MHC class II(low) cell population as a source of IL-4 in the lung after chronic allergen challenge. In the bone marrow a similar cell was identified with approximately a third of the IL-4(+) cells also expressing c-kit(+). The pulmonary and bone marrow IL-4(+) cell populations were significantly reduced upon local pulmonary anti-SCF treatment. Subsequently, when IL-25R was examined during the chronic allergen responses the expression was found on the IL-4(+) myeloid cell population that expressed CD11b(+)GR1(+). Interestingly, the IL-25R(+) cells in the bone marrow were also all CD11b(+)GR1(+), similar to the lung cells, but they were also all c-kit(+), potentially suggesting a maturation of the bone marrow cell once it enters the lung and/or is stimulated by SCF. Overall, these studies suggest a complex relationship between SCF, bone marrow-derived IL-25-responsive myeloid cells, Th2 cytokines, and chronic allergic disease.
Subject(s)
Cytokines/biosynthesis , Interleukins/biosynthesis , Lung/immunology , Myeloid Cells/immunology , Receptors, Interleukin-17/biosynthesis , Respiratory Hypersensitivity/immunology , Stem Cell Factor/physiology , Th2 Cells/immunology , Animals , Bone Marrow Cells/immunology , Bone Marrow Cells/metabolism , Bone Marrow Cells/pathology , CD11b Antigen/biosynthesis , Cell Differentiation/genetics , Cell Differentiation/immunology , Chronic Disease , Cytokines/antagonists & inhibitors , Inflammation Mediators/metabolism , Inflammation Mediators/physiology , Interleukin-17/biosynthesis , Interleukin-4/biosynthesis , Lung/metabolism , Lung/pathology , Mice , Mice, Inbred BALB C , Mice, Transgenic , Myeloid Cells/metabolism , Myeloid Cells/pathology , Receptors, Chemokine/biosynthesis , Respiratory Hypersensitivity/metabolism , Respiratory Hypersensitivity/pathology , Stem Cell Factor/antagonists & inhibitors , Th2 Cells/metabolism , Th2 Cells/pathologyABSTRACT
IL-25 (IL-17E) is a unique IL-17 family ligand that promotes Th2-skewed inflammatory responses. Intranasal administration of IL-25 into naive mice induces pulmonary inflammation similar to that seen in patients with allergic asthma, including increases in bronchoalveolar lavage fluid eosinophils, bronchoalveolar lavage fluid IL-5 and IL-13 concentrations, goblet cell hyperplasia, and increased airway hyperresponsiveness. IL-25 has been reported to bind and signal through IL-17RB (IL-17BR, IL-17Rh1). It has been demonstrated recently that IL-17A signals through a heteromeric receptor composed of IL-17RA and IL-17RC. We sought to determine whether other IL-17 family ligands also utilize heteromeric receptor complexes. The required receptor subunits for IL-25 biological activities were investigated in vitro and in vivo using a combination of knockout (KO) mice and antagonistic Abs. Unlike wild-type mice, cultured splenocytes from either IL-17RB KO or IL-17RA KO mice did not produce IL-5 or IL-13 in response to IL-25 stimulation, and both IL-17RB KO and IL-17RA KO mice did not respond to intranasal administration of IL-25. Furthermore, treatment with antagonistic mAbs to either IL-17RB or IL-17RA completely blocked IL-25-induced pulmonary inflammation and airway hyperresponsiveness in naive BALB/c mice, similar to the effects of an antagonistic Ab to IL-25. Finally, a blocking Ab to human IL-17RA prevented IL-25 activity in a primary human cell-based assay. These data demonstrate for the first time that IL-25-mediated activities require both IL-17RB and IL-17RA and provide another example of an IL-17 family ligand that utilizes a heteromeric receptor complex.
Subject(s)
Interleukin-17/physiology , Interleukins/physiology , Receptors, Interleukin-17/physiology , Receptors, Interleukin/physiology , Animals , Cells, Cultured , Humans , Interleukin-17/metabolism , Interleukins/deficiency , Interleukins/genetics , Interleukins/metabolism , Ligands , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , Rats , Rats, Inbred Lew , Receptors, Interleukin/deficiency , Receptors, Interleukin/genetics , Receptors, Interleukin-17/deficiency , Receptors, Interleukin-17/geneticsABSTRACT
The epithelial-derived cytokine thymic stromal lymphopoietin (TSLP) is sufficient to induce asthma or atopic dermatitis-like phenotypes when selectively overexpressed in transgenic mice, or when driven by topical application of vitamin D3 or low-calcemic analogues. Although T and B cells have been reported to be dispensable for the TSLP-induced inflammation in these models, little is known about the downstream pathways or additional cell types involved in the inflammatory response driven by TSLP. To characterize the downstream effects of TSLP in vivo, we examined the effects of exogenous administration of TSLP protein to wild-type and genetically deficient mice. TSLP induced a systemic Th2 inflammatory response characterized by increased circulating IgE and IgG1 as well as increased draining lymph node size and cellularity, Th2 cytokine production in draining lymph node cultures, inflammatory cell infiltrates, epithelial hyperplasia, subcuticular fibrosis, and up-regulated Th2 cytokine and chemokine messages in the skin. Responses to TSLP in various genetically deficient mice demonstrated T cells and eosinophils were required, whereas mast cells and TNF-alpha were dispensable. TSLP-induced responses were significantly, but not completely reduced in IL-4- and IL-13-deficient mice. These results shed light on the pathways and cell types involved in TSLP-induced inflammation.
Subject(s)
Cytokines/administration & dosage , Eosinophils/immunology , Eosinophils/pathology , Inflammation Mediators/administration & dosage , Th2 Cells/immunology , Th2 Cells/pathology , Thymus Gland/immunology , Thymus Gland/pathology , Animals , Cells, Cultured , Cytokines/biosynthesis , Cytokines/genetics , Cytokines/physiology , Female , Immunoglobulin Class Switching , Immunoglobulin E/biosynthesis , Inflammation Mediators/physiology , Injections, Intradermal , Male , Mast Cells/immunology , Mast Cells/pathology , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , Mice, Nude , Mice, Transgenic , Signal Transduction/genetics , Signal Transduction/immunology , Skin/immunology , Skin/pathology , Stromal Cells/immunology , Stromal Cells/pathology , Thymic Stromal LymphopoietinSubject(s)
Asthma/drug therapy , Drug Discovery/methods , Receptors, Immunologic/antagonists & inhibitors , Receptors, Prostaglandin/antagonists & inhibitors , Acetates/chemistry , Acetates/pharmacology , Acetates/therapeutic use , Animals , Asthma/immunology , Carbazoles/chemistry , Carbazoles/pharmacology , Carbazoles/therapeutic use , Clinical Trials as Topic , Humans , Indoleacetic Acids/chemistry , Indoleacetic Acids/pharmacology , Indoleacetic Acids/therapeutic use , Receptors, Immunologic/immunology , Receptors, Prostaglandin/immunology , Sulfonamides/chemistry , Sulfonamides/pharmacology , Sulfonamides/therapeutic useABSTRACT
The IL-23/IL-17 pathway is implicated in autoimmune diseases, particularly psoriasis, where biologics targeting IL-23 and IL-17 have shown significant clinical efficacy. Retinoid-related orphan nuclear receptor gamma t (RORγt) is required for Th17 differentiation and IL-17 production in adaptive and innate immune cells. We identified JNJ-54271074, a potent and highly-selective RORγt inverse agonist, which dose-dependently inhibited RORγt-driven transcription, decreased co-activator binding and promoted interaction with co-repressor protein. This compound selectively blocked Th17 differentiation, significantly reduced IL-17A production from memory T cells, and decreased IL-17A- and IL-22-producing human and murine γδ and NKT cells. In a murine collagen-induced arthritis model, JNJ-54271074 dose-dependently suppressed joint inflammation. Furthermore, JNJ-54271074 suppressed IL-17A production in human PBMC from rheumatoid arthritis patients. RORγt-deficient mice showed decreased IL-23-induced psoriasis-like skin inflammation and cytokine gene expression, consistent with dose-dependent inhibition in wild-type mice through oral dosing of JNJ-54271074. In a translational model of human psoriatic epidermal cells and skin-homing T cells, JNJ-54271074 selectively inhibited streptococcus extract-induced IL-17A and IL-17F. JNJ-54271074 is thus a potent, selective RORγt modulator with therapeutic potential in IL-23/IL-17 mediated autoimmune diseases.
Subject(s)
Arthritis, Rheumatoid/drug therapy , Nuclear Receptor Subfamily 1, Group F, Member 3/genetics , Peptides, Cyclic/administration & dosage , Psoriasis/drug therapy , Th17 Cells/drug effects , Administration, Oral , Animals , Arthritis, Rheumatoid/genetics , Arthritis, Rheumatoid/metabolism , Cell Differentiation/drug effects , Cells, Cultured , Disease Models, Animal , Dose-Response Relationship, Drug , Gene Expression Regulation/drug effects , Interleukin-17/metabolism , Interleukins/metabolism , Mice , Nuclear Receptor Subfamily 1, Group F, Member 3/metabolism , Peptides, Cyclic/pharmacology , Psoriasis/genetics , Psoriasis/metabolism , Th17 Cells/cytology , Th17 Cells/metabolism , Transcription, Genetic , Interleukin-22ABSTRACT
Thymic stromal lymphopoietin (TSLP), interleukin-25 (IL-25), and IL-33 are important initiators of type 2-associated mucosal inflammation and immunity. However, their role in the maintenance of progressive type 2 inflammation and fibrosis is much less clear. Using chronic models of helminth infection and allergic lung inflammation, we show that collective disruption of TSLP, IL-25, and IL-33 signaling suppresses chronic and progressive type 2 cytokine-driven inflammation and fibrosis. In a schistosome lung granuloma model or during chronic Schistosoma mansoni infection in the liver, individual ablation of TSLP, IL-25, or IL-33/ST2 had no impact on the development of IL-4/IL-13-dependent inflammation or fibrosis. However, significant reductions in granuloma-associated eosinophils, hepatic fibrosis, and IL-13-producing type 2 innate lymphoid cells (ILC2s) were observed when signaling of all three mediators was simultaneously disrupted. Combined blockade through monoclonal antibody (mAb) treatment also reduced IL-5 and IL-13 expression during primary and secondary granuloma formation in the lungs. In a model of chronic house dust mite-induced allergic lung inflammation, combined mAb treatment did not decrease established inflammation or fibrosis. TSLP/IL-33 double-knockout mice treated with anti-IL-25 mAb during priming, however, displayed decreased inflammation, mucus production, and lung remodeling in the chronic phase. Together, these studies reveal partially redundant roles for TSLP, IL-25, and IL-33 in the maintenance of type 2 pathology and suggest that in some settings, early combined targeting of these mediators is necessary to ameliorate progressive type 2-driven disease.
Subject(s)
Cytokines/metabolism , Fibrosis/immunology , Inflammation/immunology , Inflammation/therapy , Interleukin-17/metabolism , Interleukin-33/metabolism , Lung Neoplasms/immunology , Animals , Antibodies, Monoclonal/therapeutic use , Cytokines/antagonists & inhibitors , Cytokines/genetics , Female , Fibrosis/drug therapy , Fibrosis/therapy , Granuloma/drug therapy , Granuloma/immunology , Granuloma/parasitology , Granuloma/therapy , Inflammation/drug therapy , Interleukin-13/antagonists & inhibitors , Interleukin-13/genetics , Interleukin-13/metabolism , Interleukin-17/antagonists & inhibitors , Interleukin-17/genetics , Interleukin-33/antagonists & inhibitors , Interleukin-33/genetics , Interleukin-4/antagonists & inhibitors , Interleukin-4/genetics , Interleukin-4/metabolism , Lung Neoplasms/drug therapy , Lung Neoplasms/parasitology , Lung Neoplasms/therapy , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , Schistosoma mansoni/immunology , Schistosoma mansoni/pathogenicity , Thymic Stromal LymphopoietinABSTRACT
Interleukin-25 (IL-25) is a cytokine associated with allergy and asthma that functions to promote type 2 immune responses at mucosal epithelial surfaces and serves to protect against helminth parasitic infections in the intestinal tract. This study identifies the IL-25 receptor, IL-17RB, as a key mediator of both innate and adaptive pulmonary type 2 immune responses. Allergen exposure upregulated IL-25 and induced type 2 cytokine production in a previously undescribed granulocytic population, termed type 2 myeloid (T2M) cells. Il17rb(-/-) mice showed reduced lung pathology after chronic allergen exposure and decreased type 2 cytokine production in T2M cells and CD4(+) T lymphocytes. Airway instillation of IL-25 induced IL-4 and IL-13 production in T2M cells, demonstrating their importance in eliciting T cell-independent inflammation. The adoptive transfer of T2M cells reconstituted IL-25-mediated responses in Il17rb(-/-) mice. High-dose dexamethasone treatment did not reduce the IL-25-induced T2M pulmonary response. Finally, a similar IL-4- and IL-13-producing granulocytic population was identified in peripheral blood of human subjects with asthma. These data establish IL-25 and its receptor IL-17RB as targets for innate and adaptive immune responses in chronic allergic airway disease and identify T2M cells as a new steroid-resistant cell population.
Subject(s)
Asthma/immunology , Cytokines/biosynthesis , Interleukins/physiology , Myeloid Cells/immunology , Receptors, Interleukin-17/physiology , Adoptive Transfer , Animals , Asthma/pathology , Dexamethasone/pharmacology , Female , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Myeloid Cells/cytologyABSTRACT
Butyrophilin-like 2 (BTNL2) is a butyrophilin family member with homology to the B7 costimulatory molecules, polymorphisms of which have been recently associated through genetic analyses to sporadic inclusion body myositis and sarcoidosis. We have characterized the full structure, expression, and function of BTNL2. Structural analysis of BTNL2 shows a molecule with an extracellular region containing two sets of two Ig domains, a transmembrane region, and a previously unreported cytoplasmic tail. Unlike most other butyrophilin members, BTNL2 lacks the prototypical B30.2 ring domain. TaqMan and Northern blot analysis indicate BTNL2 is predominantly expressed in digestive tract tissues, in particular small intestine and Peyer's patches. Immunohistochemistry with BTNL2-specific Abs further localizes BTNL2 to epithelial and dendritic cells within these tissues. Despite its homology to the B7 family, BTNL2 does not bind any of the known B7 family receptors such as CD28, CTLA-4, PD-1, ICOS, or B and T lymphocyte attenuator. Because of its localization in the gut and potential role in the immune system, BTNL2 expression was analyzed in a mouse model of inflammatory bowel disease. BTNL2 is overexpressed during both the asymptomatic and symptomatic phase of the Mdr1a knockout model of spontaneous colitis. In functional assays, soluble BTNL2-Fc protein inhibits the proliferation of murine CD4(+) T cells from the spleen, mesenteric lymph node, and Peyer's patch. In addition, BTNL2-Fc reduces proliferation and cytokine production from T cells activated by anti-CD3 and B7-related protein 1. These data suggest a role for BTNL2 as a negative costimulatory molecule with implications for inflammatory disease.