Lecture 5: Complement, etc. 1. Complement a.

A family of numerous activation and regulatory proteins used as defense against invading antigens etc. i. The constituent proteins are pro-enzymes requiring activation via proteolytic hydrolysis. b. Made up of three pathways that take place in a sequential cascade i. Classical complement ii. Alternate complement iii. Terminal pathway (Development of Membrane Attack Complex [MAC]) 1. Both classical and alternate converge into this terminal pathway 2. Functions of complement a. Facilitates the interaction of phagocytes to induce opsonin-mediated phagocytosis b. Induce osmotic lysis of microbe (via insertion of MAC into membrane) c. Enhances vascular permeability by inducing degranulation of mast cells / basophils. d. Induces chemotaxis (movement) of neutrophils. e. Facilitates immune complex elimination. i. Can do this by binding C3b to 1. Complexes in circulation eliminated by neutrophils. 2. Transport complexes to liver/spleen via RBCs. a. RBCs have CR1 so they can bind C3b. C3b serves as an opsonin in this situation. 3. Classical complement a. Initiated using IgG or IgM i. Remember IgM is the most potent activator ii. The Fab portion of the antibody is bound to the molecule and the Fc region is free to bind complement proteins. b. C1 binds to the free Fc portion. i. C1 is made up of C1q, C1r, and C1s. c. C1 cleaves C4 to C4a and C4b. i. C4b binds to C1. ii. C4a is a free anaphylatoxin. d. C1 cleaves C2 to C2a and C2b. i. C2a binds to C1. ii. C2b is a weak kinin that increases vascular permeability. e. C4b + C2a constitutes C3 convertase. i. C3 convertase cleaves C3 to C3a and C3b. 1. C3a is a free anaphylatoxin. 2. C3b binds to C3 convertase forming a. C5 convertase. f. C4b + C2a+ C3b = C5 convertase. i. C5 convertase cleaves C5 to C5a and C5b.

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1. C5a is a free anaphylatoxin. 2. C5b joins with C6, C7, C8, and C9 to form MAC. What are anaphylatoxins? These are molecules that bind to receptors on mast cells and basophils and induce their degranulation. This results in the release of histamine and other inflammatory mediators. a. C4a is only produced in the classical pathway. b. C3a and C5a are both anaphylatoxins. Alternate Complement a. C3 Tickover i. Small amounts of spontaneously generated complement fragments, C3b, occur with or without an infection. 1. If a microbe is present it is used to activate alternate complement. 2. If a microbe is not present water or complement regulatory proteins inactivate it. b. Initiated by C3b binding to the surface of a microbe. i. Factor B rapidly deposits once this happens. 1. Factor D cleaves B to Ba and Bb. c. C3b + Bb = Alternate complement C3 convertase. i. This molecule is stabilized by properdin. d. C3 convertase cleaves a C3 to C3a and C3b. i. C3bBbC3b = Alternate complement C5 convertase. e. C5 convertase cleaves C5 and continues to the terminal pathway. Classical pathway regulators a. C1 INH i. Complexes with C1 preventing spontaneous activation of classical complement. ii. Also inactivates kallikrein, a protease that links complement with the coagulation pathway. b. Decay Accelerating Factor (DAF) i. Binds C4b and C3b blocking C3 convertase formation. 1. Blocks classical AND alternate complement. c. C4bp i. When C4b is free it prevents it from attaching. ii. Competitively binds C4b promoting dissociation. d. Factor I i. Cleaves C4b to an inactive form, C4bi. e. Anaphylatoxin inhibitor (AI) i. Binds C3a, C4a, and C5a inhibiting them from binding to receptors on mast cells and basophils preventing degranulation. 1. For classical and alternate complement Alternate pathway regulators a. Factor H i. When free it prevents C3b from binding to cell surface.

ii. When in alternate pathway C3 convertase if formed it competitively binds C3b inducing dissociation. b. DAF i. Same as above c. AI i. Same as above d. Factor I i. Cleaves C3b to the inactive form C3bi. 8. Terminal Pathway inhibitors a. These prevent formation of MAC i. Homologous Restriction factor (HRF), S-protein, and CD59. b. These prevent MAC insertion into membranes by binding solube [C5b, C6, and C7]. i. S-protein/vitronectin c. Binds C8 preventing polymerization to C9. i. CD59 / HRF20 9. Kallikrein and the intrinsic pathway [*This is a for interest section*] a. When tissue is damaged Factor XII is activated. i. Activated Factor XII is called Factor XIIa. b. In the presence of kinninogen XIIa cleaves pre-kallikrein to kallikrein. i. Kallikrein enhances the response by converting XII to XIIa providing more for the coagulation process. c. Kallikrein i. Can use C5 as a substrate producing C5a and C5b. ii. Cleaves kinninogen to kinninogen a and bradykinin. 1. Bradykinin is a potent vasodilator. Addendum: Classical Complement The Stanley Method