Allergic diseases and treatment. Feng Qian ( 钱峰 )
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1 Allergic diseases and treatment Feng Qian ( 钱峰 ) fengqian@sjtu.edu.cn
2 Hypersensitivity Antigen Host Adaptive responses protective to the host harmful to the host The deleterious consequences of the adaptive responses to the host cells
3 Characteristics of hypersensitivity Excessive immune response in a sensitized individual leading to tissue damage. Adaptive immunity is important against microbial infections, but it is also capable of causing tissue injury and disease. (autoimmune diseases)
4 Characteristics of hypersensitivity Occurs when immune responses are directed against self-ag and also from uncontrolled or excessive responses to against foreign Ag, such as microbes and allergens Common cause is failure of self-tolerance, which ensures that individuals do not respond to their own antigens Leads to tissue injury that occurs in autoimmune diseases due to the same effector mechanisms used to protect against microbes
5 Types of hypersensitivity Type I: IgE antibodies bind to Fc receptors on mast cells. IgE induces mast cell degranulation and release inflammatory mediators Type II: Ab (IgG/IgM) mediated immune response against self antigen or foreign antigen (ie Ag on transfused RBC) Type III: Ab(IgG/IgM)+Ag=Immune complexes are deposited in tissue Type IV: T-cell mediated response where Ag sensitized T-cells release lymphokines
6 Types of hypersensitivity
7 Types of hypersensitivity
8 Type I hypersensitivity IgE antibodies bind to Fc receptors on mast cells. IgE induces mast cell degranulation and release inflammatory mediators Occur and resolve quickly Mediated by serum IgE Systemic and regional tissue dysfuntion Genetic predisposition
9 Type I hypersensitivity 1) Priming stage:last more than half a year 2) Activating stage: Crosslinkage Enzyme reaction Degranulation 3) Effect stage: Immediate/early phase response: Mediated by histamine Start within seconds Last several hours Late-phase response : Mediated by new-synthesized lipid mediators Take up 8-12hours to develop Last several days
10 Components and cells in type I hypersensitivity Allergen : pollen dust mite insects etc selectively activate CD4+Th2 cells and B cells Allergin(IgE)and its production IgE: mainly produced by mucosal B cells special affinity to the same cell IL-4 is essential to switch B cells to IgE production High affinity receptor of the IgE on mast cell and basophil Eosinophil
11 IgE-mediated responses Fc R Fc RI (high affinity ) Fc RII (low affinity ) (CD23) Lysis scd23(ige-bf) Fc RIIa Fc RIIb on circulating basophil, connective tissue mast cells on B cell on B, T, Mφ, DC, basophil Switch B cell to IgE production
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14 Mast cell activation by IgE crosslinking 14
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16 The biological mediators 1. Histamine: Dilate blood vessel Increase vascular permeability 2. Leukotrienes: Bronchial smooth muscles contract Asthma 3. Prostaglandin: High concentration of PGE low concentration of PGE 4. Platelet activating factor (PAF) : Agglutinate and activate platelets to release Inhibit the secretion of histamine promote the release of histamine histamine 5. Eosinophil chemotactic factor(ecf-a): 6. Bradykinin : Vasodilator function
17 Mechanism of type I hypersensitivity Allergen Primary Individual Generation IgE Secondary Adhesion IgE binds to the Fc RI on mast cell and basophil Allergen binds to the IgE on primed target cell Crosslikage of Fc RI Degranulate and release the biological mediators Preformed granule mediators New generated mediators Histamine Bradykinin Leukotrienes PAF Prostaglandin D2 Dilate capillaries,increase permeability, increase mucus secretion, contract smooth muscle Systemic anaphylaxis Skin Respiratory tract Degist tract
18 Common diseases of type I hypersensitivity 1. Systemic anaphylaxis: a very dangerous syndrome 1) Anaphylactic drug allergy (penicillin) 2) Anaphylactic serum allergy 2. Respiratory allergic diseases : 1) Allergic asthma:acute response, chronic response 2) Allergic rhinitis 3. Gastrointestinal allergic diseases : The lack of SIgA protein hydrolase 4. Skin allergy: Undigested protein Allergen
19 Asthma Asthma is a common clinical condition, characterized by variable airflow obstruction, occurring both spontaneously and in response to nonspecific triggers such as exercise, smoke, and fumes. Chronic lung condition Reversible reactive airway obstruction Tightening of the muscles surrounding the bronchial passages in the lungs Can be developed at any age
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21 Epidemiology of asthma million people worldwide are affected by asthma thousand people die per year from asthma by 2011
22 Asthma symptoms Coughing especially at night, Wheezing Dyspnea (shortness of breath) Chest tightness Chronic inflammatory disorder of the airways Airway obstruction and airway hyperresponsiveness
23 Common asthma triggers Extrinsic Dust mites Mold Pollen Certain foods Animal dander Intrinsic Exercise Infections Cold/humid air Stress/anger Medications Hormones Air pollution Fragrances Chemicals Cannot be cured wikipedia
24 Immune responses in asthma Eosinophil Macrophage CD4+ T cell IL-5 IL-4? Asthma Allergen IL-13 Macrophage Dendritic cell B cell Ig E Mast cell
25 Ovalbumin-induced asthma model i.p. (OVA+Alum) i.n. (OVA or PBS) Time (day) Analysis Egg OVA Structure Allergy Asthma Immunol Res. 2010, 2:
26 DRA-induced asthma model Dust mite Ragweed Aspergillus Acute model DRA-induced mouse acute asthma model i.p. (DRA+Alum) i.n. (DRA or PBS) Analysis Time (day) DRA-induced mouse chronic asthma model Chronic model 1 i.n. (DRA) Time (week) Analysis 12
27 DRA-induced asthmatic responses BAL total cell counts (X10 5 ) ** * 0 DRA ( g) Siglec F PBS 1.0 Eos MФ CD-11c DRA 84.8 Eos MФ H&E staining Computerized analysis
28 Transcription Factor PU.1 PU.1 is a key differentiation regulator (3000 genes) in hematopoietic cells. Higher level in macrophages, intermediate in B cells, little in T cells. PU.1 modulates B cells, dendritic cells, eosinophils, and macrophages. PU.1 promotes neutrophilic lung inflammation.
29 PU.1 +/- mice displayed attenuated DRAinduced asthmatic inflammation DRA-induced mouse acute asthma model i.p. (DRA+Alum) i.n. (DRA or PBS) Analysis Time (day) DRA PBS WT PU/ER(T) +/- Histology WT PU/ER(T) +/- PBS DRA Inflammatory cell infiltration
30 PU.1 +/- mice displayed attenuated DRAinduced asthmatic inflammation WT WT PU/ER(T) +/- PU/ER(T) +/- 1.0 Eos Mac 73.8 Eos Mac DRA PBS Eos Mac Eos Mac 25.5 DRA Siglec F PBS CD-11c Cytospin Flow cytometry
31 PU.1 +/- mice displayed attenuated DRAinduced asthmatic inflammation +/- 5 Th2 responses PU.1 promotes allergic asthmatic inflammation
32 Macrophage polarization Macrophage M1 Ym-1, Fizz-1, arginase-1, IL-10, MR,SR, GR M2 Classical activation IL-1β, IL-12, IL-23, TNFα, inos Alternative activation
33 Roles of M2 macrophage in diseases Alternatively activated macrophage host defense against parasite wound healing diabetes by contributing to insulin resistance tumorogenesis allergic inflammation
34 Structure of respiratory system Trachea Epithelium O 2 CO 2 Endothelium
35 Macrophage Fas-Induced Apoptosis (MAFIA) Transgenic mice Csf1r promoter GFP IRESmembrane-anchored Fas death domain FKBPs Vehicle MAFIA Fas-ligand Fas death domain FKBPs Fasligand AP20187 % of Max Macrophage or Dendritic cell Apoptosis GFP
36 Depletion of macrophages alleviates DRA-induced asthmatic inflammation i.p. (DRA) i.p. Fas-ligand i.n. (DRA) Assay Days BAL total cells (X10 4 ) DRA Fas-ligand ND ND
37 Depletion of macrophages alleviates DRA-induced asthmatic inflammation PBS DRA Vehicle Fas-ligand Periodic-acid-Schiff (PAS) stain Macrophage is required for asthma.
38 M2 peritoneal macrophages i.p. IL-4( 2 ug) Days -3-1 Collection of PM PBS IL-4 Arginase 1/GAPDH (fold) CCL22 mrna (fold) PBS IL-4 In vivo injection of IL-4 triggers M2 macrophage polarization.
39 Adoptive transfer of M2 macrophage accentuates asthmatic inflammation Recipient Dono Transfer of M2 macrophages i.p. ( low dose of r DRA) i.n. (DRA) Assay Day s % Eos Adoptive Transfer (i.t. injection) Mac IL-4-treated Mac 4% 87% 1% MФ Eos MФ ** * 6 5 Siglec F 17% CD-11c 3% 9% 2% 0 0 PBS-Mac IL-4-Mac
40 Adoptive transfer of M2 macrophage promotes asthmatic inflammation Mac WT IL-4-treated Mac WT M2 macrophages enhances asthma.
41 Adotive transfer of WT macrophages rescues asthmatic inflammation Adoptive transfer of macrophages i.p. IL 4( 2 g) Donor i.t. injection of PMs Days 3 i.p. (DRA Alum) i.n. (DRA) Assay Recipient Days PU/ER(T) +/- PU/ER(T) +/- WT PU/ER(T) +/- BAL total cell counts (X10 5 ) 9 * DRA Eosinophils (x10 5 ) *** DRA
42 Adotive transfer of WT macrophages rescues asthmatic inflammation Donor PU/ER(T) +/ Mac Recipient PU/ER(T) +/ Mice Eos Mac WT Mac PU/ER(T) +/ Mice Eos Mac DRA Siglec F CD-11c DRA Eosinophil acumlation Periodic-acid-Schiff (PAS) stain PU.1 promotes asthmatic inflammation via macrophages
43 DRA-induced M2 phenotypes are attenuated in the lung tissue of PU.1 +/- mice WT PU/ER(T) +/- WT PU/ER(T) +/- WT PU/ER(T) +/ PBS DRA 0 PBS DRA 0 PBS DRA RT-PCR PU.1 dificient mice shown attenuated M2 macrophage polarization
44 IL-4-mediated signal pathways Type I IL-4R IL-4 c IL-4R Type II IL-4R IL-4/13 IL-13R 1 IL-4R Jak3 Jak1 Tyk2 Shp-1 Y Y Y Y IRS Shc Stat6 A Y Y Y Jak1 Y Stat6 PU.1 P P P Stat6 PU.1Stat6 IRF4 PPAR Arg1, Fizz, Ym-1, CCL17/22
45 DRA-induced M2 phenotypes are attenuated in the lung tissue of PU.1 +/- mice IL-4 (5 ng/ml) Time (min) STAT6-Pi STAT6 PU.1 GAPDH STAT6-Pi/STAT6 15 ** 10 ** ** Time (min) PU.1/GAPDH ** * Time (min) IL-4 induces PU.1 expression that is associated with Stat6 activation.
46 PU.1 promotes M2 macrophage polarizaiton Ym-1/GAPDH (fold) WT PU/ER(T) +/- PBS ** IL-4 Fizz-1/GAPDH (fold) WT PU/ER(T) +/- ** PBS RT-PCR IL-4 Arginase-1/GAPDH (fold) WT PU/ER(T) +/- PBS IL-4
47 PU.1 promotes M2 macrophage polarizaiton Ym-1 GAPDH Fizz-1 GAPDH PBS +/+ +/- 5 +/+ +/- IL-4 (ng/ml) 10 +/+ +/- Ym-1/GAPDH WT PU/ER(T) +/- * * PBS 5 10 IL-4 (ng/ml) Fizz-1/GAPDH (fold) WT PU/ER(T) +/- *** ** PBS 5 10 IL-4 (ng/ml) Transcriptional factor PU.1 is required for Ym-1 and Fizz-1 expression.
48 PU.1 modulates M2 macrophage polarization IL-4 Extracellular IL-4Rα Jak3 Jak1 PU.1 P P P PU.1? Fizz-1 Ym-1 Arginase-1 Transcription
49 Genotypic characterization of PU/ER(T) mice Blood 2011
50 PU/ER(T) [PU.1/Estrogen recepor] mice Inactivation Activation ER ER PU.1 Tamoxinfen ER ER PU.1 ER ER PU.1 Fizz-1/Ym-1 ER ER PU.1 Transcription
51 Reconstitution of functional PU.1 leads to increased asthmatic inflammation BAL cell counts (X10 5 ) Placebo Tamoxifen *** * ** * PBS DRA Eosinophils (x10 5 ) Placebo Tamoxifen ND/ND PBS ** DRA Macrophage (x10 5 ) Placebo Tamoxinfen PBS DRA
52 Reconstitution of functional PU.1 leads to increased asthmatic inflammation Siglec F Eos (21%) CD11c Placebo Mac (5.2%) DRA Eos (69%) Tamoxifen Mac (1.9%) IgE (ng/ml) Placebo Tamoxifen ** PBS DRA To confirm that PU.1 promotes asthmatic inflammation.
53 PU.1 contributes to DRA-induced chronic asthmatic inflammation DRA-induced mouse chronic asthma model 1 i.n. (DRA) Time (week) Analysis 12 BAL cell counts (X10 5 ) WT PU/ER(T) +/- ** PBS DRA Eosinophils (x10 5 ) WT PU/ER(T) +/- *** ND/ND PBS DRA Macrophages (x10 5 ) WT PU/ER(T) +/- PBS * DRA Inflammatory cell infiltration
54 PU.1 contributes to DRA-induced chronic asthmatic inflammation Ym-1/GAPDH (fold) WT PU/ER(T) +/- * PBS DRA Fizz-1/GAPDH (fold) WT PU/ER(T) +/- PBS ** DRA Arginase-1/GAPDH (fold) WT PU/ER(T) +/- PBS * DRA M2 macrophage polarization (RT-PCR)
55 PU.1 contributes to DRA-induced chronic asthmatic inflammation Raw H&E Genie tissue classification WT PU/ER(T) +/-
56 PU.1 contributes to DRA-induced chronic asthmatic inflammation WT PU/ER(T) +/- PBS DRA Periodic-acid-Schiff (PAS) stain
57 Role of PU.1 in asthmatic inflammation Th2 Cell IL-4 Asthma IL-4/13R IL-13 PU.1 Jak1/3 CCR4 CCL17 CCL22 IL-13 Fizz1 P Stat6 Alternative Activation of Macrophage P PU.1 IRF4 PPAR Fizz1 Eosinophil
58 PU.1 promotes asthma via modulating macrophage polarization IL-4 Macrophage PU.1 Jak1 Y Jak3 Y Stat6 Stat6 P Stat6 PU.1 Stat6 P Ym-1, Fizz- 1 JMCB, 2015
59 Summary Types I hypersensitivity Pathogenessis of asthma Macrophage is required for asthma Alternatively activated macrophages (M2) promote asthma Transcriptional factor PU.1 plays a critical role in macrophage polarization and asthmatic inflammation
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