Complement & Complement Fixation Hugh B. Fackrell 3/17/2018 1
Content Outline l l Complement Components Complement Activation Biological Consequences Complement Fixation 3/17/2018 2
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Complement system 3/17/2018 6
Phases in the Complement Cascade Recognition l Activation Amplification l Membrane Attack l 3/17/2018 7
Complement Cascade 3/17/2018 8
Complement Activation Classical pathway l Alternative pathway l Membrane attack complex l 3/17/2018 9
Complement Components The Classical Pathway 3/17/2018 10
Membrane Bound Antigens 3/17/2018 11
Soluble Antigen 3/17/2018 12
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C 4 3/17/2018 15
C 2 b 3/17/2018 16
C 3 3/17/2018 17
C 5 3/17/2018 18
C 5 a & C 5 b 3/17/2018 19
C 6 3/17/2018 20
C 7 3/17/2018 21
C 7 becomes hydrophobic 3/17/2018 22
C 8 3/17/2018 23
C 8 in membrane 3/17/2018 24
C 9 3/17/2018 25
C 9 pore 3/17/2018 26
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An Alternative Complement Pathway Part of Innate immunity Activated without Antibody 3/17/2018 28
Alternate Vs Classical Paths 3/17/2018 29
Regulation of Complement 3/17/2018 30
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Biological Consequences Cell lysis l Inflammation l Opsonization l Viral neutralization l Solubilization of immune complexes l 3/17/2018 32
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Complement Fixation Test Fig 18. 8 3/17/2018 36
Complement Fixation Assay Test Phase l Positive serum – antibodies l l l Negative serum – no antibodies Antigen Complement No Ag-Ab complex Complement NOT used up Antigen l Complement l AG-AB complex l formed l Complement used up No visible difference!!! 3/17/2018 37
Complement Fixation Assay Indicator phase l Positive Test phase l – No complement l Indicator system – Complement l – Sheep RBCs – Anti sheep RBCs l Result – NO LYSIS of RBCs – CLOUDY 3/17/2018 Negative test phase Indicator System – Sheep RBCs – Anti Sheep RBC’s l Result – LYSIS of RBCs – CLEAR 38
CF: Advantages Reagents easily available l Semi quantitative or quantitative l Wide application l Extremely sensitive l 3/17/2018 39
CF: Disadvantages Indirect Test l Need adequate controls l Reagents unstable l Quantitation time consuming l Cannot be automated l 3/17/2018 40
DONE!!! 3/17/2018 41
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