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Recognition of microbes by innate immune system A. Specificity innate has limited diversity compared with adaptive 1. Compontesn f innate immunity recognize structures shared by various classes of microbes (molecular patterns) and not present on host cells 2. Pathogen associated molecular patterns (PAMP) microbial molecules that are targets of innate a. Phagocytes express receptors for bacterial LPS (endotoxin) present in cell wall of many bacterial species but not produced by mammalian cells b. Terminal mannose of glycoproteins, - typical in bacteria c. Ds RN found in many viruses d. Unmethylated CpG oligonucleotides, common to bacterial DNA B. Receptors: cellular receptors of innate recognize structures that are essential for survival/infectivity 1. Microbe cannot evade innate by mutating or not expressing targets of recognition a. Adaptive immunity- microbes evade by mutating antigens recognized by lymphocytes 2. Receptors recognize molecules relapsed from stressed or necrotic cells, and results in elimination of these cells a. Damage associated molecular patterns (DAMPs) b. Non infectious inflammatory response C. Receptors are encoded in the germline and lead to limited diversity 1. Lymphocytes receptors encoded by recombination of receptor genes which leads to greater diversity 2. Innate receptors are nonclonally distributed identical receptors on all cells of the same lineage a. Adaptive has clonal distribution where clones of lymphocytes with distinct specificities express different receptors D. Discrimination of self and nonself innate does not react against the host 1. Inherent specificity of innate for microbial structures 2. Mammalian cells have regulatory molecules that prevent innate immune reactions E. Memory does not demonstrate in innate 1. Responds in same way to repeat encounters with a microbe 2. Adaptive response responds more efficiently to after repeated infection with same microbe F. Two principal types of reactions of innate 1. Inflammation recruitment and activation of leukocytes 2. Anti viral defense mediated mainly by NK cells and cytokines (interferons) Cellular receptors for microbes cell types including phagocytes, lymphocytes, dendritic, epithelial and endothelial express receptors for innate expressed at different cellular compartments (cell surface, ER endosomes, cytoplasm) A. Toll like receptors (TLR) 1. Homologous to Drosophila protein called Toll 2. Different TLRs respond to different products of microbes 13 different in mammals a. Each TLR either by itself or in combination with other TLRSs

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3. Activate similar signaling mechanisms, resulting in responses critical for innate response B. Specified and functions of TLRS 1. TLR 1, 2,6 a. PAMPs bacterial peptidoglycan, lipoprotein b. PAMPs pathogen associated molecular patterns 2. TLR 4 a. PAMP gram negative bacterial LPS; fungal mannas; viral envelope proteins 3. TLR -5 a. PAMP bacterial flagella 4. Binding of ligand to TLR activates TF called NF-kB which produces cytokines Components of innate immunity A. Epithelial barriers include skin (physical contact), gastrointestinal tract (ingestion) and respiratory tract (breathing) 1. Continuous epithelia provide physical and chemical barriers against infection 2. Prevent microbe entry by 3 mechanisms a. Physical barrier b. Killing of microbes by locally produced antibiotics (peptide) i. Ex: defensins and crptocidisn (intestine) c. Killing of microbes and infected cells by intraepithelial lymphocytes B. Phagocytes: neutrophils and monocytes/macrophages 1. Two types of circulating phagocytes: neutrophils and monocytes that re recruiting to the site of infection, where hey recognize and ingest microbes for intracellular killing a. Neutrophils (polymorphonuclear leukocytes) most abundant (400-10000 per u>=L) i. Production of neutrophils from bone marrow increases rapidly in response to infections ii. Cytokines secreted by cells in response to infections and act on BM stem cells to stimulate proliferation and maturation of neutrophils iii. First on scene, but die after few hours b. Monocytes ingest microbes in blood and tissue i. Bone marrow precursors lead to mononuclear phagocytes, which exits during circulation in the blood ii. In the tissues monocytes become macrophages iii. Macrophage are either activated by microbes or may differentiate into specialized forms depending on the tissue 2. Sequence of events of leukocyte migration a. Migrate to sites of infection by binding to endothelial adhesion molecules and in response to chemoattracts that r produced upon encounter with microbes b. Resident macrophages and dendritic cells ingest microbes and produce cytokines

c. Cytokines active endothelial cells of blood vessels to produce selectins, ligand for integrins and chemokines d. Surface cab of circulating neutrophils and monocytes bind weakly to selectin which slows the down blood flow pushes them resulting in the rolling of cells on endothelial surface e. When rolling integrins expressed on surface of leukocytes bind to integrin ligand on epithelial cells i. Endothelial cells produce chemokines which bind go glycoproteins on luminal surface of endothelium, which increases affinity of leukocyte integrins of ligands ii. Firm binding of integrins to ligands stops rolling leukocytes on endothelium f. Chemokines stimulate motility of leukocytes which being migrate between endothelial cells to site of infection g. Accumulation of leukocytes at site of infection along with vascular dilation and permeability is called inflammation h. Leukocyte adhesion deficiencies: inherited deficiencies in integrins and selectin ligands lead to defective leukocyte recruitment and increased susceptibility to infection 3. 4 steps a. Rolling, mediated by selectins b. Activation by chemoattractant stimulus c. Arrest and adhesion mediated by integrins binding to Ig family d. Transendothelial migration 4. Neutrophils and macrophages uses several types of receptors to recognize microbes and to initiate responses that destroy microbes 5. Leukocytes phagocytosed microbes and destroy ingested microbes in intracellular vesicle phagocytosis a. Microbe binds to phagocyte receptor and membrane zips up around the microbe, internalizing microbe -> phagosome b. Phagosome fuse with lysosome to produce phagolysosome c. Killing of microbes by lysosomal enzymes in phagolysosomes i. ROI reactive oxygen intermediates such as superoxide anion and free radicals ii. No nitric oxide d. After phagocytosis, he remaining debris is eliminated from the cell by exocytosis C. Dendritic cells respond to microbe b y producing cytokines that recruit leukocytes and initiate adaptive immune response D. Natural killer cells class of lymphocytes that recognize infected and stressed cells and respond by killing these cells and by secreting macrophage activating cytokine IFN-g 1. Activation of NK macrophages that have encountered microbes active NK cells by producing cytokines (IL12) 2. NK cells response a. Discharge proteins contained in their cytoplasmic granules toward infected cells i. Some proteins create holes in PM of infected cells and other enter the infected cell and induce apoptosis

b. Synthesize IFN -gamma which activates macrophages to become more effective in killing microbes 3. Interaction of class 1 Major histocompatibility complex (MHC) molecules with inhibitory receptor of NK prevents killing of healthy host cells by NK cells a. Healthy cells express self class 1 MHC b. NK cells respond to the absence of MCH class 1 cells E. Other calluses of lymphocytes F. Complement system composed of several proteins that circulate in blood stream 1. Activation of system initiates cascade that aids in destruction of invading organism a. Helps induce inflammatory response 2. Many proteins are proteolytic enzymes and complement activation involves sequential activation of these enzymes 3. Pathways of complement system a. Alternative pathway triggered when complement proteins activated on microbial surface and cant be controlled i. Activated by microbes in absence of antibodies ii. Parte of innate immunity b. Classical pathway triggered after antibodies bind to microbe s c. Lectin pathway 0 activated when PP, mannose binding lectin, binds to terminal mannose residues on the surface glycoproteins of microbes i. Lectin activates protein of classical pathway ii. Still part of innate immunology because it initiated by microbial product and in the absence of an antibody 4. Serves 3 functions in host defense a. C3b coats microbes and promotes binding of microbes to phagocytes i. Microbes that are opsonized with complement proteins are rapidly ingested and destroy b. Proteolytic fragments of complement protein, C5a and C4a are chemoattractant for phagocytes and promote leukocyte recruitment at the site of complement activation c. Complement activation culminates in formation of a polymeric protein complex that inserts in microbial cell membrane which disturbs permeability barrier and causing osmotic lysis G. Cytokines of innate immunity dendritic cells, macrophages and other cells secrete cytokines that mediate many of the cellular rxns of innate 1. Soluble proteins that mediate immune and inflammatory reactions and responsible for communications between leukocytes and other cells 2. Most are interleukins molecules produced by leukocytes and act on leukocytes 3. Mechanism of cytokine action a. Bind to specific high affinity receptors on target cells b. Can be Autocrine or paracrine

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c. Cytokine when bound to tis recepotr activate specific signaling pathways in cytoplasm resulting in activation of TF in nucleus that alter gene expression 4. Functions of innate immunity to a. Stimulate inflammation (TNF, IL1, chemokines) b. Activate NK cells (Il-12) c. Activate macrophages (IFN-g) d. Prevent viral infections (type I IFNs) H. Other plasma proteins of innate immunity involved in defense against infections 1. Plasma mannose binding lectin (MLB) binds to bacterial carbohydrates to target them for phagocytosis s a. Activate complement cascade by the lectin pathway 2. C reactive protein (CRP) a. Targets microbes for phagocytosis by macrophages b. Circulating levels of many of these proteins increase rapid after infection c. Protection by other plasma proteins is called acute phase response Evasion of innate immunity by microbes A. Evasion of innate immunity by microbes 1. Pneumococcus resistance to phagocytosis a. Capsular polysaccharide inhibits phagocytosis 2. Staphylococci resistance to reactive oxygen intermediates in phagocytes a. Mechanism: production of catalase which breaks down RO intermediates 3. Neisseria meningitides resistance to complement activation (alternative pathway) a. Mechanism sialic acid expression inhibits C3 and C5 convertases 4. Streptococcus resistance to complement activation (alt. pathway) 5. Pseudomonas - Resistance to antimicrobial peptide antibiotics a. Synthesis of modified LPS resistant action of peptide antibiotics Role of innate immunity in stimulation adaptive responses A. Mechanism by which innate immunity stimulate adaptive immune responses 1. Innate immune response has a warning function to warn adaptive immune system and provide second signals for he activation of B and T lymphocytes 2. Two signals required for activation of lymphocytes a. Ensure lymphocytes respond to pathogenic infectious microbes and not to harmless materials 3. Generate molecules to function as second signal together with antigen to activate T and B lymphocytes a. Signal 1: antigen, component of a microbe b. Signal 2: co stimulatory molecule produced or induced by microbes 4. Nature of second signal stimulate adaptive immunity and guides nature of adaptive immune response

a. Intracellular and phagocytosed microbes need to eliminated by cell-mediated immunity (T lymphocytes) i. Microbes stimulate dendritic ells and MP to produce two types of second signals ii. Dendritic and MP express surface molecules called costimulators which bind to receptors on nave T cells and function with antigen to activate T cells iii. Dendritic cells and MP secrete cytokine IL 12 Stimulates diff. of nave T cells into effector cells of cell mediated adaptive immunity] b. Blood borne microbes need to combated by antibodies which need activation of B lymphocytes, the humoral immune response i. Blood borne microbes activate plasma complement system (alternative pathway) which, along with antigen recognition, stimulates B cell differentiation into antibody secreting cells ii. Complement product serves as second signal for humoral immune response 5. Different type of microbes induce different innate immune response, which then stimulate the types of adaptive immunity that are best t to combat different infectious pathogens