Ch4: B Cells Flashcards
Define immunoglobulin
proteins which structurally resemble antibodies.
Define antibodies
proteins made by lymphocytes and plasma cells, which specifically react with molecules termed antigens (Ags).
All antibodies are what type of protein?
Immuoglobulins
Where did the name antigen come from?
ANTIbody GENerator
Define antigen
any compound that elicits an immune response
What is an immunogen
Antigen that causes immune response
What is immunogenicity dependent on? (3)
foreignness, size, and complexity
What two types of molecules induce strong immune response?
Proteins and polysaccharides
Recognition of antigen directed at what?
antigenic determinant or epitope
What is an epitope?
The chemical structures of a molecule that is directly recognized
Antibodies are composed of what?
heavy and light chains that both contain variable and constant regions
Antibodies are made up of how many chains?
2 identical heavy chains
2 identical light chains
Antigens are bound to what part of antibody?
What holds the antigen in place?
fragment antigenbinding (Fab) region Disulfide bonds
What part of the antibody tells the host how to handle the antigen:antibody complex?
The fragment crystallizable (Fc) region
The interactions of antibodies and antigen are mediated by what types of forces?
1) electrostatic forces,
2) hydrogen bonds,
3) Van der Waals forces,
4) hydrophobic forces
Since the interactions are less stable between antigens and antibodies (non-covalent) what does this allow the antibody to do?
bind multiple sites on the pathogen
How many sites can each antibody bind?
IgM: 10 IgA: 4 IgG: 2 IgE: 2 IgD: 2
Linear epiotopes can be predicted by what?
Amino acid sequence
Discontinuous epitomes can be predicted by what?
Amino acid sequence AND 3-D folding knowledge
IgA has how many subtypes?
2
What is the heavy chain of IgA?
Alpha (1 or 2)
Secreted form of IgA?
Monomer or Dimer
Function of IgA?
Mucosal immunity
H chain of IgD?
Delta
Secreted form of IgD?
Isn’t one
Function of IgD?
Native B cell antigen receptor
H chain of IgE?
Episilon
Secreted form of IgE?
Monomer
Function of IgE? (2)
- Mast cell activation
2. Defense against helminthic parasites
H chain type of IgG?
Gamma (1,2,3,4)
Secreted form of IgG?
Monomer
Function of IgG? 5
- Opsonization
- Complement activation
- Antibody-dependent cell-mediated cytotoxicity
- Neonatal immunity
- Feedback inhibition of B cells
IgM H chain type?
Mu
Secreted form of IgM?
Pentamer
Function of IgM? (2)
- Native B cell antigen receptor
2. Complement activation
B cells are produced where?
By what cell?
Bone marrow
Hematopoietic stem cell
The germline DNA contains multiple copies of what types of genes that will be rearranged? 3
variable (V), diversity (D), and joining (J) genes
Why does gene rearrangement occur for B cells?
provide significant variability to the antigen-binding portion of immunoglobulins
The heavy chain uses what germline genes?
VDJ
The light chain uses what germline genes?
VJ
Successful rearrangement of the VDJ (heavy chain) and VJ (light chain) allows these genes to do what?
associate with constant genes, and the rearrangement is complete
Diversity in T and B cells means what?
The number of antigen specificities present within a single individual at any given time
B cell germline rearrangements occur when?
Throughout life
Do we have immunoglobulins for antigens we have not encountered?
Yes
How many Variable gene segments exist for light chains?
Kappa: 31-36
Lambda: 29-33
How many variable gene segments for heavy chain?
38-46
How many diversity segments in light chains?
0
How many diversity segments in heavy chains?
23
How many joining segments in light chains?
Kappa: 5
Lambda: 4-5
How many joining segments in heavy chains?
6
How many constant segments in light chains?
Kappa: 1
Lambda: 4-5
Which gene rearrangements occur first, light or heavy?
Heavy (D to J) then (V to DJ)
Gene rearrangements for light chain involve what segments?
V to J fusion
Each V, D, or J segments is flanked by what?
Recombination Signal Sequences (RSS)
Two types of Recombination Signal Sequences
- Nonamer and Heptamer separated by a 12 base pair segments
2. Nonamer and heptamer separated by a 23 bp segments
The 12/23 rule ensures what?
Gene segments are joined in the correct orientation
During rearrangement of gene segments, what two genes bind to the 12 bp and 23 bp spacers?
Function of this?
RAG–1 and RAG-2
Bring two heptamer sequences in close proximity through formation of a hairpin that contains the DNA segments to be removed
DNA is excised where in germline rearrangment?
End of both heptamer sequences
What joins chromosomal DNA together following recombination?
Terminal deoxynucleotidal transferase (TdT)
Describe the steps of Germline rearrangement 8
- Generation of junctional diversity
- RAG complex cleaves heptamer RSS’s from the D and J gene segments to yield hairpins
- RAG complex opens hairpins by nicking one strand of DNA which generates palindromic P-nucleotides
- N-nucleotide additions are performed by TdT
- TdT adds N nucleotides
- Pairing of strands occurs
- Unpaired nucleotides are removed by exonuclease
- Gaps are filled by DNA synthesis and ligation to form coding joint
What are P nucleotides?
pallindromic nucleotides that are identical when read from either end, but they are not complementary to each other and cannot join together.
What are N nucleotides?
Nontemplated/not encoded in germline DNA
How does TdT contribute to diversity of antibodies?
Formation of coding joint involves TdT adding N-nucleotides
How might two B cells with identical V, D, and J genes differ?
Different N-nucleotides added
After B cells rearrange the heavy chain, they express what?
For what reason?
Express a Pre-B cell receptor
To test the ability to express the heavy chain at the cell surface
What is needed to make the Pre-B cell receptor?
- Heavy Chain
- Surrogate light chain
- Two disulfide linked signaling molecules (Igalpha and IgBeta)
B cells that possess an appropriate heavy chain in the Pre-B cell receptor undergo what?
cell division to produce around 100 clones expressing the same heavy chain
Do clones of B cells have the same light chain?
No. each clone begins making light chains independently
How many chances does a B cell have to create a proper light chain?
Why?
4
both κ light chain and λ light chain gene rearrangements are possible
If a successful light chain rearrangement does not occur, what happens to the B cell?
Dies by apoptosis
If a successful light chain rearrangement does occur, what happens?
the B cell receptor is expressed at the cell surface along with two disulfide-linked signaling molecules (Igα and Igβ)
As an early pro-B cell, what can the B cell do for rearrangements?
D-J rearrangements on both chromosomes
As a late pro-B cell, what can the B cell do for rearrangements?
And if this fails?
And if this fails?
- V-DJ rearrangement on first chromosome
- V-DJ rearrangement on second chromosome
- Apoptosis
If successful rearrangement occurs of V-DJ, what can the B cell do next for light chain rearrangement? If this fails? If this fails? If this fails? If this fails?
- Rearrange Kappa gene on first chromosome
- Rearrange Kappa gene on second chromosome
- Rearrange Lambda gene on first chromosome
- Rearrange lambda gene on second chromosome
- Apoptosis
If successful kappa light chain rearrangement occurs in Pre-B cell, what is the result?
Cell expresses IgM of mu/Kappa
If successful lambda light chain rearrangement occurs in Pre-B cell, what is the result?
Cell expresses IgM of mu/Lambda
What are the two checkpoints in determining whether a B cell will make functional IgM?
- First: Can it make a pre-B cell receptor
2. Second: Can it make a mature B cell receptor
What is allelic exclusion?
process of encoding immunoglobulin genes from a single chromosome per cell, instead of both
What is affinity?
Strength of binding at a single site
What is avidity?
Strength associated with binding multiple
sites, regardless of affinity
Having multiple B-cell receptors that recognize the same antigen increases what?
increases the avidity and potential for activation of the B cell because multiple signals tell B cell to become activated
After the V, D, and J genes rearrange, the B cell can proceed to rearragnements with what genes?
Constant genes
How many constant genes exist on heavy chain of B cells?
What do they determine
9
define the antibody isotype that is being produced
by the cell.
The default constant gene for all B cells is what? (2)
Cμ (IgM) and Cδ (IgD)
B cell express what antibodies at cell surface during development?
IgM and IgD
What two types of IgM do B cells express?
surface IgM and secreted IgM
Downstream constant genes (not the default ones) exist for what reason?
Future isotype switching
How many different V gene segments exist for B cell heavy chains?
100
How many different V gene segments exist for B cells’ kappa chains?
35
How many different D gene segments exist for B heavy chains?
27
How many different D gene segments exist for B kappa chains?
0
How many different J gene segments exist for B kappa chains?
5
How many different J gene segments exist for B heavy chains?
6
Which has the better possible repertoire of antigen diversity, TcR’s or Immunoglobulin?
TcR’s 10^16 vs 10^11
The primary transcript for antibody is processed by what 3 things?
cleavage, polyadenylation, and splicing
Processing of primary antibody transcript creates what? 2
- μ transcript that does not encode the δ,
2. δ transcript that does not encode μ.