Lecture 6: Antibodies and Antigens Flashcards
Antibody structure
2 heavy chains
2 light chains
Chains bound together by disulfide bond & non covalent interactions
Chains have constant and variable regions
V and C regions
V region contains Ab binding site
C region determines fate of Ab
Antibody cleavage
Antibodies can be cleaved to yield two Fab fragments (Fragment Ag binding site) and an Fc fragment (responsible for effector functions)
Cleavage by papain gives
Two Fab fragments and an Fc fragment
Cleavage by pepsin
Generates single bivalent Ag binding fragment F(ab’)2
Heavy chains and their Ig’s
u (mu) - IgM S (Delta) - IgD y (gamma) - IgG e (epsilon) - IgE a (alpha) - IgA
Light chains
k (kappa)
l (lambda)
Ab unit can only have one, not both
H - L chain bonds
Both chains have intrachain disulfide bridges every 90 AAs, creating polypeptide loops
Ag binding site in secreted IgG formed by
Juxtaposition of Variable light and Variable heavy domains
Difference b/w membrane bound IgM and secreted IgG
Membrane IgM has one extra CH4 domain
Conformational determinant
Only recognizes folded, non-denatured proteins
Linear determinant
If there is an accessible linear sequence, Ig can bind determinant in native form. If not, it must be denatured
Neoantigenic determinant
Proteolysis creates a site that can be bound
Affinity of Abs formed in the primary response vs Memory response
Lower affinity in primary, higher in memory response
When is affinity critical
If Ag is toxin or Virus and must be eliminated immediately at low titers
Ab valence
Maximum number of antigenic determinants with which it can interact
IgG valence
IgG has two Fab regions and can bind two molecules of Ag - valence is 2
Avidity and what it is dependent upon
Gives a measure of the overall strength of an Ab-Ag complex
Dependent on affinity of Ab for epitope and valence of both Ab and Ag
Immature and mature B cells contain what Ig
IgM on immature
IgM and IgD on mature
Changes in Ig after activation of B cells
Low rate of Ig secretion at first
Heavy chain isotype switching, affinity maturation
Then becomes Ab secreting cell w/high rate of Ig secretion and reduced membrane Ig
Affinity maturation occurs when and where
Only occurs after activation, which would only occur in secondary lymphoid organs
Ig with secretory component
Only IgA
IFN-y signal to B cell
Signals B cell to become plasma cell and secrete IgG for phagocytosis
TGF-B signal to B cell
Signals B cell to become plasma cell and secrete IgA for mucosal defense
IL-4 signal to B cell
Signals B cell to become plasma cell and secrete IgE
IL-2, 4, 5 signal to B cell
Signals B cell to become plasma cell and secrete IgM, causing complement activation
IgA found
Mucosal secretions
IgM found
Mostly in plasma
IgE found
Absorbed on surface of mast cells
IgG found
Penetrates deeper into the tissue
IgG structure/function
Provides bulk of immunity for blood-born pathogens
Two heavy y(gamma) chains and either two Kappa or Lambda light chains
What results in different subclasses of IgG
Slightly different sequences in their H-chains
IgG and babies
IgG is the only Ab to cross the placenta and provide passive humoral immunity to developing fetus
This will last for a couple weeks after birth
FcRn
Cell surface receptor for Fc region of IgG in placenta, allows passing into fetal circulation
IgD function
Low quantities in circulation
It is an Ag receptor on B lymphocytes
Naive B cells express
Both IgM and IgD, which are specific for the same Ag
If Ag binds, it is internalized and presented to helper T cells
IgM function
First Ab produced by Ag activated B cell
Plays critical role in defense before sufficient IgG has been synthesized
Found primarily in blood stream
High avidity, making IgM effective in the removal of the microbe
IgM structure
Five four-chain units held together by disulfide bonds
J-chain initiates polymerization of subunits during secretion from plasma cell
IgA function/synthesis
Major Ig found in external secretions
Most synthesized by plasma cells in mammary/saliva glands in respiratory, GI, GU tracts
Major defense mechanism against microbes at mucosal surfaces
Secreted IgA structure
In addition to L and H chains, also contains secretory component (SC) and and J-chain
Secretory component (SC) function
Part of the Ig receptor involved in the transepithelial transport of exocrine IgA and stabilizes IgA against degradation in GI tract
IgE function
Very low serum levels
Significant role in enhancing acute inflammation and protein from infection by worms and allergies
IgE interaction with mast cells
IgE binds to receptors on mast cell that recognize Fc portion of Ig
When Ag is introduced to a mast cell with IgE, it triggers activation and release of mediators