1、固有免疫系统固有免疫系统固有屏障结构固有屏障结构皮肤与粘膜皮肤与粘膜血脑屏障血脑屏障胎盘屏障胎盘屏障固有免疫分子固有免疫分子补体补体溶菌酶溶菌酶防御素防御素固有免疫细胞固有免疫细胞PMNDCsMf fvIntact skin/Mucus membranevMucosal immune system (MIS)vSecretion of anti-microbial factorsvNormal flora vBlood-brain and placenta barrierslSkin and mucosal barriervPhysical: skin & mucosa, epidermis
2、-dermis conjunction, mucosal secretion, lavaging of fluids and ciliary actionvChemical: acidic secretion, lysozymevMicroorganism: commensal bacteriaBlood-thymus-barrierBlood-gas-barrierBlood-urine-barrierBlood-testis-barrierBlood-epididymis-barrierCerebral barrier: BBB, etc.Blood-fetus-barrierMucosa
3、-bicarbonate barriervType of phagocytes: vSmall phagocytes (neutrophils) vlarge phagocytes (macrophage) vDendritic Cells vChemotaxis Contact Phagocytosis Intracellular killingvPhagocytic effects enhanced by antibody or complement l吞噬和杀菌过程：吞噬和杀菌过程：趋化趋化黏附黏附 吞入吞入 杀灭与消化杀灭与消化(CD14)1.Complment: alternativ
4、e & MBL pathway2.Lysozyme: tissue and body fluid, secre-tion, lysosome in phagocytes3.Antimicrobial peptides: defensin4.Cytokines: interferon, IL-1, IL-6, IL-12, TNF-, chemotaxin5.Acute phase protein: CRP, MBP, SAP, 1-acid glycoprotein, FN, etc.1.Killing G+ bacteria, splits the exposed peptidoglycan
5、 wall2.Category: 3.Bacterial lysozyme4.Act on -1,4 glycosidic bond5.Act on the peptide tail and amide6.Fungal lysozyme: Saccharomycetic, Aspergillar1.Interferon: virally infected cells can synthesize type IFN. Reduce mRNA translation, degrade mRNA, inhibit viral replication2.Interleukin: IL-1, IL-6,
6、 IL-123.Chemotactic factors4.TNF: TNF-Father of Innate Immunity：Prof. Janeway CConcept:PRR : Pathogen recognizing receptorsPAMPs: Pathogen associated molecular patterns（一）识别什么？（一）识别什么？PAMPs1、PAMP: Pathogen Associated Molecular PatternsLip polysaccharide (LPS), dsRNA, unmethylated CpG, mannose, gluca
7、n, Lipoteichoicacid (LTA), Pepidoglycan (PGN)2、PRR: receptors on professional APC and M which enable them to recognize PAMP. mannose receptorToll-like receptorscavenger receptorphosphatidyl serine receptor29uPAMP are conserved, shared by a large group of infectious agents, and clearly distinguishabl
8、e from self patterns.uTypical PAMP: lipopolysaccharide(LPS), mannose, glucan, dsRNA, lipoteichoicacid(LTA), peptidoglycan(PGN) unmethylated CpG DNA etc黏附素（黏附素（adhesin)adhesin)fimbrial adhesins (E. coli )non-fimbral adhesin （Bartonella）黏附素黏附素（二）用什么去识别？（二）用什么去识别？PRR3636Expressed mainly on DC and Mf wh
9、ich enable them to recognize PAMP. Three types of PRR:secreted PRR: MBL,C-reactive proteinendocytic PRR: scavenger receptor, mannose receptorsignaling PRR: Toll-like receptor, NOD-like receptor, RIG-like receptor3737Drosophila embryogenesis. (1980s)Antifungal response in Drosophila adults. (Hoffmann
10、 et al., Cell, 1996)A human homologue of Toll (Janeway et al., Nature, 1997)3838 Jules A. Hoffmann Bruce BeutlerThey received one-half of , for their discoveries concerning 43uRecognize cytoplasmic PAMPs and/or endogenous danger signals, inducing immune responses.uLigands: MDP and meso-DAPuThree dom
11、ains:C-terminal LRRs sense ligands centrally located NOD domainN-terminal effector region comprising a protein-protein interaction domain such as the CARD, Pyrin or BIR domain.uSeveral subfamilies (on the basis of their effector domains)CARD subfamily: NODs ,IPAF, CIITA,PYD subfamily: NALPsBIR subfa
12、mily: NAIPs5253uThey may then form oligomers that activate inflammatory caspase1 causing cleavage and activation of important inflammatory cytokines（IL-1）uNLRs may also activate the NF-kB signaling pathway to induce production of inflammatory molecules.5555u inflammasome, a multiprotein complex that
13、 contains, in addition to NLR, the adaptor molecule ASC and the protease caspase-1.u inflammasome is responsible for the proteolytic processing of the immature form of the cytokines belonging to the IL-1 family (IL-1b, IL-18, and IL-33), 5657Fabio Martinon，Annu. Rev. Immunol. 2009. 27:229uDetect int
14、racellular dsRNA of viral origin:RIG-I (retinoic acid induced gene)MDA-5 (melanoma differentiation associated gene)LGP2 (laboratory of genetics and physiology)uComprises N-terminal caspase activation and recruitment domains (CARDs), a DECH helicase, and a C-terminal domain (CTD).60Red box, CARD1; bl
15、ue box, CARD2; green box, Helicase domain; purple box, DEAD/DEAH domain6163The Journal of Clinical Investigation, 118(2), 200869视频视频 (Immune Response)O2-dependent: ROI, RNI (reactive nitrogen intermediate), SOD (superoxidase dismutase)O2independent: MPO (myeloperoxidase, ) Enzymes (lysozyme) Defensi
16、ns 热热 红红 肿肿 痛痛 功能丧失功能丧失 炎症炎症CEDFBAHuang X. and Hazlett LD. (2003) IOVS 44:3409-3416 3d5d030006000900012000 00 BALB/cB601020304050601 day p.i.NormalATLR9 mRNA levels TLR9 IL-1b b MIP-20102030405060GpC controlCpG treatmentBRalative mRNA levelsg g0102030405060CRelative mRNA levels0102030405060CpG treat
17、mentGpC controlD01 02 Real-time PCR of fold changes (normalized to b-actin) in TLR9, IL-1b and MIP-2 mRNA expression in XS52 LC treated with TLR9-siRNA and CpG-ODN. Treatment TLR9 IL-1b b MIP-2 CpG-ODN (control) 7.8 225.6 237.5 Control-siRNA+CpG 5.1 148.2 157.1 TLR9-siRNA+CpG 1.4 53.5 41.2 03 ABCD04
18、C g gControl-siRNATLR9-siRNADTLR9-siRNAControl-siRNAEAB05 ABC0602000400060008000A0400080001200016000B050100150200250C0246810DLog10CFU/cornea0708Huang X et al, IOVS, 2005; 46:4209-4216 （第一和通讯作者：黄曦）（第一和通讯作者：黄曦）Huang X et al, IOVS, 2006; 47:4910-4916 （第一和通讯作者：黄曦）（第一和通讯作者：黄曦）09101112IOVS, 2007; 48:4626-
19、4633 （第一作者：黄曦）（第一作者：黄曦）J Immunol. 2006; 177:548-556（通讯作者：黄曦）（通讯作者：黄曦）13141516N12h1 d3 d5 d020406080100p0.001p0.001p0.01p=0.23p=0.41Days postinfectionSIGIRR mRNA levelsBALB/cC57BL/617ADays postinfection012345 IgG controlAnti-SIGIRR Ab1d3d5d7dClinical scoreBCDE183d5d024681012 IgG ControlAnti-SIGIRR Ab
20、p0.001p=0.007Days postinfectionLog10 CFU/cornea191d3d5d0200040006000800010000 IgG controlAnti-SIGIRR Abp0.001p0.01Ap=0.26Days postinfectionIL-1b b protein (pg/ml)1d3d5d030006000900012000150001800021000 IgG controlAnti-SIGIRR Abp0.001p0.001p=0.03BDays postinfection MIP-2 protein (pg/ml)201d3d5d020406
21、080100120 IgG controlAnti-SIGIRR AbAP0.001P0.01P=0.12Days postinfectionRelative IL-1R1 mRNA levels1d3d5d0255075100 IgG controlAnti-SIGIRR AbBP=0.016P=0.01P=0.67Days postinfectionRelative TLR4 mRNA levels21 1d3d5d020406080100120 Bp0.05p0.01p=0.58Days postinfectionRelative IFN-g g mRNA levels1d3d5d025
22、5075100 Ap0.01p0.01p0.01Days postinfectionRelative IL-18 mRNA levels221d3d5d0255075100 IgG controlAnti-SIGIRR AbAp=0.94p=0.86p=0.58Days postinfectionRelative IL-4 mRNA levels1d3d5d0255075100 IgG controlAnti-SIGIRR AbBp=0.42p=0.89p=0.93Days postinfectionRelative IL-10 mRNA levels23B SIGIRR mRNA expre
23、ssionNormal6 h18 h060120180240300 LPS HK-PAASIGIRR mRNA levelsEmpty vectorRAW264.7 cellRAW264.7 cellSIGIRR-vectorSIGIRR-vectorEmpty vectorC SIGIRR protein levels Empty vectorEmpty vectorPositive controlSIGIRR-vectorSIGIRR-vectorSIGIRR24IL-1R1TLR-4TLR304080120160200Empty vectorSIGIRR vectorAp0.01p0.0
24、01p=0.83Relative mRNA levelsIL-12IL-18IFN- g g050100150200250Empty vectorSIGIRR vectorBp0.01p0.001p0.01Relative mRNA levels25AIL-5IL-10020406080100Empty-vectorSIGIRR-vectorp=0.64p=0.78Relative mRNA levelsIL-1b bMIP-2100101102103Empty vectorSIGIRR vectorBp0.01p0.01Relative mRNA levelsCProtein levels (pg/ml)IL-1b bMIP-2100101102103104105106Empty vectorSIGIRR vectorp0.01p0.012627282903060901201501803031323334353637IL-1b bMIP-2IL-6TNF-a a38394041
