Professional Documents
Culture Documents
B Cells and B Cell Development: Dr. Mansour Elyazji
B Cells and B Cell Development: Dr. Mansour Elyazji
B Cells and B Cell Development: Dr. Mansour Elyazji
Bursa was later found to be the organ in which antibody producing cells
developed – antibody producing cells were thereafter called B cells
Mammals do not have a bursa of Fabricius
Origin of B cells and organ of B cell
maturation
Mature marked
Transfer marked foetal B cells
liver cells in periphery
Normal bone marrow
No Mature
B cells
B
Regulates construction of an antigen receptor
S
M
E
Scheme of B Cell Development in the Bone Marrow
Immature &
Progenitors Pre-B mature B
X
Central
E Sinus
n
d
o
o
s
t
X
e
u
m
Stromal cells
X
Macrophage
Bone marrow stromal cells nurture
developing B cells
Secreted
Cell-cell contact Factors - CYTOKINES
B
Stromal cell
Stromal cell
Stages of B cell development
Stem Cell Early pro-B cell Late pro-B cell Large pre-B cell
Peripheral
Kit
Early Receptor
VLA-4 Stem pro-B Tyrosine
(Integrin) kinase
Stem cell
VCAM-1
(Ig superfamily)
factor
Cell-bound
growth
Cell adhesion factor
molecules
Stromal cell
Cytokines and cell-cell contacts at each
stage of differentiation are different
Interleukin-7
receptor
Interleukin-7
Growth factor
Late
Early pro-B Pre-B
pro-B
Stromal cell
Stages of differentiation in the bone marrow are
defined by Ig gene rearrangement
B CELL STAGE Stem cell Early pro-B Late pro-B Large pre-B
IgH GENE
Germline DH to JH VH to DHJH VHDHJH
CONFIGURATION
Pre-B cell
receptor
expressed
Ligand for the pre-B cell receptor may be galectin 1, heparan sulphate, other
pre-BCR or something as yet unknown
Ligation of the pre-B cell receptor
ALLELIC EXCLUSION
Expression of a gene on one chromosome prevents expression of the
allele on the second chromosome
Allelic exclusion is needed for efficient clonal selection
Antibody
S. typhi S. typhi
Y Y
AND
Y
anti-S. aureus Ig
Y
Y
B B
Exclusion of anti-brain B cells
i.e. self tolerance BUT anti S.aureus B cells
will be excluded
B B leaving a “hole in the
OR
repertoire”
Deletion Anergy
Y Y
Y
S. aureus
B
Ligation of the pre-B cell receptor
Large Large
Pre-B
Large
Pre-B
Large Many large pre-B
Large Proliferation
Large
Pre-B
Large
Pre-B
Large
Pre-B
Large
Pre-B cells with identical
Large Large
pre-B Pre-B
Pre-B
Pre-B
Pre-B
pre-B receptors
Y
Small
Large Immature
pre-B
IgM
B cell
Proliferation
Intracellular VDJCH chain Light chain expressed
stops
Pre-receptor VL-JL rearranges IgM displayed on surface
not displayed
Heavy and light chain rearrangement is potentially wasteful
V D J C Germline
V J C Germline
V V J C VL-JL joining
Y
Y
B BY
Small pre-B cell Immature B cell
No antigen receptor at cell surface Cell surface Ig expressed
Unable to sense Ag environment Able to sense Ag environment
!!May be self-reactive!! Can now be checked for self-reactivity
Small
B
B
pre-B Immature
YY BB
Small
YY Y IgM
Y
Y
pre-B Immature IgD IgD
BB YB
IgD
B
Small pre-B cell Immature
assembles Ig B cell recognises
soluble self Ag
No cross-linking
Anergic B cell
Receptor editing
A rearrangement encoding a self specific receptor can be replaced
V V V V D J C
!!Receptor
Y
B recognises
self antigen!!
Arrest development
And reactivate B Apoptosis
or anergy
RAG-1 and RAG-2
V V V D J C
Small
YY IgM
IgD
YY IgM
B Immature
YY
pre-B
BB IgD
YY
IgD
YYB
IgM IgM
IgD
Mature B cell
Small pre-B cell Immature exported to the
assembles Ig B cell doesn’t periphery
recognise any
self Ag
How can B cells express
IgM and IgD simultaneously?
S3
C C
C3
C C3 C S1
C1
VD J C C C3 C1
V D J C IgM mRNA
RNA cleaved and
polyadenylated at pA2
VD J C1 C2 C3 C4 C1 C2 C3 AAA
pA1
V D J C IgD mRNA
Summary
Extracellular antigen
recognition domains
Ig Ig
The cytoplasmic domains of the Ig and
Ig contain Immunoreceptor Tyrosine
-based Activation Motifs (ITAMS) - 2
tyrosine residues separated by 9-12
amino acids - YXX[L/V]X6-9YXX[L/V]
Intracytoplasmic
signalling domains
Regulation of Src kinases
CD21
(C3d receptor)
CD45
CD19
Ig Ig
CD81
(TAPA-1)
C3d binds to
CD21, the
complement
Antigen receptor 2
recognition (CR2)
• mIg and CD21 are cross-linked by antigen that has activated complement
• CD21 is phosphorylated and receptor-associated kinases phosphorylate CD19
• Phosphorylated CD19 activates more Src family kinases
• Ligation of the co-receptor increases B cell receptor signalling 1000 -10,000 fold
Transmission of signals from the cell
surface to the nucleus
YY
YY
YY B
Y
B
YY
YY
Y
B
YY
Y Y YY
YY
Y YY
Mature peripheral B cell recognises Ig-secreting plasma cell
B cell non-self antigen
in periphery
Plasma cells
B
Low No Yes No No
Plasma cell No
Summary
B cell
area
Efferent
lymph
Recirculating B cells are trapped by foreign
antigens in lymphoid organs
B cells leave blood &
enter lymph node via
B cells high endothelial venules
proliferate
rapidly
Antigen enters
node in afferent
lymphatic Y
YY
Y
YYY
YY
Y
Y
YYY
Y Germinal centre
YY
Y releases B cells
GERMINAL CENTRE that differentiate
Transient structure of into plasma cells
Intense proliferation
B cells (stained brown) in the Germinal Centre
FDC surface
DC veils
Anti- B cell
Iccosomes bearing
different antigens
Y
Y Y B
Surface Ig captures antigen CD40
Cross-linking of antigen receptor activates B cell
Activated B cell expresses CD40
Fate of Antigens Internalised by B cells
1. Capture by antigen
specific Ig maximises
B uptake of a single antigen
B
3. Antigen enters exogenous antigen
processing pathway
BYYY Th
Th
1. T cell antigen receptor
2. Co-receptor (CD4)
Signal 1
antigen & antigen 3.CD40 Ligand
receptor
Signal 2
T cell help - Signal 2
Cytokines
IL-4
IL-5
B Th IL-6
IFN-
YYY TGF-
Cytokines
Signal 1
B cells are inherently prone to die by apoptosis
Signal 1 & 2 upregulate Bcl-XL in the B cell and
Bcl-XL prevents apoptosis
CDR2
CDR3
CDR2
CDR3
CDR1
CDR3
CDR1
CDR1
CDR1
CDR2
CDR3
Only this cell, that has a high affinity for antigen can express CD40.
Only this cell can receive signal 2
Only this cell is rescued from apoptosis i.e. clonally selected
Y
Y
YYY
Y Germinal centre
YY
Y releases B cells
GERMINAL CENTRE that differentiate
Transient structure of into plasma cells
Intense proliferation
1. Antigen loaded dendritic cells
migrate from subcapsular
sinus to paracortical area
B cells (90%) and T cells
of the lymph node
(10%) migrate to form
a primary follicle
Primary follicle
B B
formation
B T 3. T cells
B B proliferate T
B DC
2. T cells migrate
B B through HEV and
T
B T T are trapped by
B T T T antigen on DC
B B T
4. B cells migrate through
HEV - most pass through the B
paracortex and primary follicle. HEV
Some interact with T cells and
proliferate to form a primary focus
T cell motility in the lymph node
Germinal Centre Microanatomy
2. B cells (centrocytes) upregulate
surface Ig, stop dividing and 4. Selected cells leave lymph
receive costimulatory signals node as memory
from T cells and FDC cells or plasma cells
Follicular dendritic
cells select useful
B cells 1. B cells (centroblasts) downregulate
3. Apoptosis of surface Ig, proliferate, somatically
self-reactive & hypermutate their Ig genes.
unselected cells AFFINITY MATURATION
Two B cell lineages
B B PC
B2 B cells
Y
?
YYYY
Y
Y
YY IgG
CD5
?
B B Y
Y B1 B cells
Y
Y Y
Distinct B cell
Y
Y
Y
Y Y
precursor
IgM - no other isotypes
B-1 B Cells
Y
Y
Y
Y
Y
Specificity & requirement for T cell help suggests strikingly different types
of antigens are seen by B-1 and B-2 B cells
T Independent Antigens (TI-2)
TI-2 Antigen
Y
Immature
B-2 Cell
Y
YY Mature
Y
B-1
Y
Y
Y Y
Y
Y Y
Y
Y Y
Y
Y
Y Y
Y
Y Y
Immature B cells that bind to
Y
Y
Y
Y Y
multivalent self Ag undergo
apoptosis IgM
Non-bone marrow derived B-1 cells
B-2 cell repertoire is purged are directly stimulated by antigens
of cells recognising containing multivalent epitopes.
multivalent antigens during
No T cells are necessary
development in the bone
marrow
Induces the expression of natural
antibodies specific for TI-2 antigens
T Independent Antigens (TI-1)
CD14
Activation of B cell
B Cell
T Independent Antigens (TI-1 e.g. LPS)
LPS complexes with CD14, LPSBP & TLR4
B B B B B B
Y Y Y Y Y Y
Six different B cells will require 6 different antigens to activate them
At high dose TI-1 antigens (like LPS) will POLYCLONALLY ACTIVATE all of
the B cells irrespective fo their specificity.
TI-1 antigens are called MITOGENS
YY YY YY YY YY YY
YY YY YY YY YY YY
Y YY YY YY YY YY Y
T Dependent & Independent Antigens
T
Dependent TI-1 TI-2
Antigens Antigens Antigens
Induce responses in babies Yes Yes No
Induce responses in athymics No Yes Yes
Prime T cells Yes No No
Polyclonally activate B cells No Yes No
Require repeating epitopes No No Yes
Why are babies unresponsive to TI-2 antigens?
In adults: TI-2 Antigen
Y
Immature
B-2 Cell
Y
YY Mature
Y
B-1
Y
Y
Y Y
Y
Y Y
Y
Y Y
Y
Y
Y Y
Y
Y Y
Adult immature B cells that
Y
Y
Y
YY
bind to multivalent self Ag
undergo apoptosis IgM
Adult non-bone marrow derived B-1
cells are directly stimulated by
antigens containing multivalent
epitopes produce IgM WITHOUT T
cell help.
Why are babies unresponsive to TI-2 antigens?
In babies:
All B cells, B-1 & B-2, are immature
TI-2 Antigen
Immature
B-1 Cell
YY
Immature B cells that bind to
multivalent self Ag undergo
apoptosis
As with adult B cells, immature B cells that bind multivalent self Ag
undergo apoptosis
Hence babies do not respond to TI-2 antigens.
Babies are, therefore susceptible to pathogens with multivalent
antigens such as those on pneumococcus
T Dependent & Independent Antigens
T
Dependent TI-1 TI-2
Antigens Antigens Antigens
Induces response in babies Yes Yes No
Induces response in athymia No Yes Yes
Primes T cells Yes No No
Polyclonally activates B cells No Yes No
Requires repeating epitopes No No Yes
Examples
TD: Diptheria toxin, influenza heamagglutinin, Mycobacterium tuberculosis
TI-1: Bacterial lipopolysaccharides, Brucella abortis
TI-2: Pneumococcal polysaccharides, Salmonella polymerised flagellin
Immune effector mechanisms against
extracellular pathogens & toxins
NEUTRALISATION
Toxin
Y
Bacterium
`
Y
`
`
` `
Adhesion to
host cells blocked
Toxin release
blocked
Y
`
Y
`
Prevents Prevents
invasion toxicity
NEUTRALISING ANTIBODIES
Effector mechanisms against
extracellular pathogens
OPSONISATION
Bacteria in extracellular space
+
Ab
+
Ab &
COMPLEMENT