Final Exam Flashcards
isotype switching
antigen binding stays same
effector cell changes
ALWAYS starts w/IgM
occurs by class switch recombination in heavy chain (Constant region)
constant regions loop out and switch regions recombine
During maturation (after activation), B cell cuts off IgM constant region and pastes on another
Pro-inflammatory cytokines
NFkB- DEpendent transcription
IL-1, TNFa, IL-6
IL-8, IL-11, IL-12, IL-15, IL-18
Anti-inflammatory cytokines
NFkB- INdependent transcription
IL-10, TGFb
Th1 cytokines
differentiated by macrophages (IFNg) and IL-12
secrete IFNg, IL-2, TNFb
Th2 cytokines
differentiated by IL-4
secrete IL-4, IL-5, IL-10
auto
self
iso
same/identical
allo
different
xeno
foreigner/alien
PGI
vasodilation
PGE
permeability
LTB4
neutrophils come B4 macrophages
Th1 isotype switch
OPSONIZATION: IgG
Th2 isotype switch
NEUTRALIZATION: IgE (allergies, parasites)
Th1 transcription factor
IFNg receptor
T-bet (IFNg)
You BET IFNg is gonna be on the test
Th2 transcription factor
GATA 3 (IL 4, 5, 13)
Th17 transcription factor
RORgT (IL 17, 22)
My husband Rory is 17, but he wants to be 22.
Innate imbalance towards inflammatory cytokines
local cytokines spill over into systemic-> systemic inflammation-> septic shock
Sepsis caused by cytokines in circulation (not necessarily the pathogen)
Severe sepsis-> cardiovascular collapse (intravasc coag) and multiple organ failure -> death
Systemic Inflammatory response syndrome (SIRS)
due to overwhelming inflammation by innate imm sys cytokines (TNFa, IL-1, IL-6, IL-12)
leads to early death
Compensatory Anti-inflammatory response syndrome (CARS)
strong inflammatory response (cytokine storm) can inc susceptibility to future infections persistent immunosuppression (IL-10, IL-4, TGFb) and recurrent infections lead to late deaths
Pro-inflammatory cytokine TIMELINE
0 hr: TNF
2 hr: IL-1
4 hr: IL-6
Lack of TNF allows pathogen to proliferate before other cytokines attack
TNF/IL-1 local effects
Vascular endothelium: leukocyte adhesion molecules, IL-1/chemokines, procoag (INFLAM)
Leukocytes: activation, cytokines (INFLAM)
Fibroblasts: proliferation, collagen synth (REPAIR)
TNF/IL-1 systemic effects
Fever Leukocytosis Acute phase proteins Sleep decreased appetite (INFLAM, "sickness syndrome")
IL-12/18/IFNg synergy
activate macrophages and NK cells
IL-12/18: macrophage binds to NK
IFNg: NK kills phagocytosed microbes in macrophage
Classically activated macrophage (M1)
TLR ligands/IFNg: Monocyte-> Macrophage ->
1. ROS, NO, lysosomal enzymes: phagocytosis/killing of microbes
2. IL-1, IL-12, IL-23, chemokines: inflam (inh by IL-10, TGFb)
Inh by IL-4, IL-13
Alternatively activated macrophage (M2)
IL-4, IL-13: Monocyte-> Macrophage -> 1. IL-10, TGFb: anti-inflam 2. Proline, polyamines, TGFb: wound repair Inh by TLR ligands, IFNg granulomas
Th1 phenotypes
Cytokines: IFNg Immune rxn: M1 activ, IgG prod Against: INTRAcellular microbes Disease: autoimmune diseases, tissue damage (from chronic infections) TF: T-bet
Th2 phenotypes
Cytokines: IL-4, 5, 13
Immune rxn: mast cell/eosinophil activ, IgE prod, M2 activ
Against: helminthic parasites (EXTRAcellular digestion)
Disease: allergic diseases
TF: GATA-3
Th17 phenotypes
Cytokines: IL-17, 22 Immune rxn: neutrophilic/monocytic inflam Against: EXTRAcell bacteria, fungi Disease: autoimmune/inflam diseases TF: RORgT
IgM
default B cell product
pentamer
complement activ
FIRST antibody that appears when an antigen is encountered for the first time (usually the one to activate B cells)
important for encapsulated bacteria
low affinity interaction enhanced by 5 adhesion sites
POTENT COMPLEMENT ACTIVATOR/OPSONIN
IgG
B cell + Th (CD40L, cytokines) + IFNg
monomer
Fc-dependent phagocytosis, complement activ, placental transfer of immunity
Neutralizes bacterial toxins and opsonizes bacteria
Neonatal immunity
IgE
B cell + Th (CD40L, cytokines) + IL-4
monomer
immunity against helminthic parasites
mast cell degranulation (immediate hypersensitivity)
IgA
B cell + Th (CD40L, cytokines) + mucosal cytokines (TGFb, BAFF)
dimer (or monomer)
proteolysis-resistant (GI tract)
mucosal immunity
Th1-mediated immunity
- APC + microbes -> Naive CD4+ T cell
- T cell prolif/diff -> Th1
- IFNg: M1 activation, B cell complement binding/opsonizing abs
- Abs + Fc receptor -> Opsonization and phagocytosis
Th2-mediated immunity
- APC + microbes/protein antigens -> naive CD4+ T cell
- Prolif/diff -> Th2 cells
- IL-4: B cells -> IgE
IL-5: eosinophils activation - IgE -> mast cell degranulation
CD8+ T cells
- CD8+ Tc cells recognize Ag + costim on APC -> CTL diff (w/o Th cells)
- CD4+ Th cells -> cytokines -> CTL diff
- CD4+ Th cells (CD40/CD40L) inc APC stimulation of CTL diff
CD8+ Tc cell killing mechanisms
Apoptosis caused by…
- Exocytosis: Granzyme and Perforin released on target cell after binding
- Fas-L-Fas (Fas-L on activated CTL, CD95 on target cell) binding (w/MHC-CD8/TCR binding)-> activates caspases (induces apoptosis)
gamma/delta T cells
DEFENSE against infection/sterile stress 1. cytokine/chemokine production 2. lyse infected/stressed cells 3. regulate stromal cell fxn w/growth factors 4. DC maturation 5. ab T cell priming w/Ag presentation 6. B cell help, IgE production FOR MY MEMORY: CK Lykes Strong, able, DC BoyEs
NKT cell antigen recognition
semi-invariant Va14-Ja18 TCR binds glycolipid on CD1d (on APC)
Cellular immunity (effector cells, pathogen location, Ag presentation, action)
Effector cells: CD8+ CTLs
Pathogen location: cytoplasm
Ag presentation: MHC I
Action: induced apoptosis
Effector cells: Type 1 CD4+ T lymphocytes
Pathogen location: MP phagolysosome
Ag presentation: MP MHC II
Action: activated pathogen killing
Humoral immunity (effector cells, pathogen location, Ag presentation, action)
Effector cells: Type 2 CD4+ T lymphocytes
Pathogen location: extracellular
Ag presentation: Pro-APC MHC II
Action: Abs prod by plasma cells
AA
Arachidonic Acid
COX
CycloOXygenase enzyme
rich in fibroblasts, smooth muscle, epithelial, endothelial, and hematopoietic cells
GSH
reduced glutathione
H1 and H2
Histamine receptors
IL
InterLeukin
IgE
Immunoglobulin E
LPS
LipoPolySaccharide
LT
LeukoTriene (LTA4, LTB4, LTC4, LTD4, LTE4)
made by leukocytes
have conjugated triene in structure
5-LO
5-LipOxygenase (enzyme)
Substrate: AA (and related PUFA)
Fxn: convert AA + O2 -> LTA4
rich in myeloid cells (macrophages, mast cells, basophils, neutrophils; inducible in monocytes)
Self-reliant chemotaxis: made by neutrophils to attract more neutrophils
NSAID
Non-Steroidal Anti-Inflammatory Drug
PG
ProstaGlandin (PGE2, PGF2, PGI2)
PGI2
ProstaGlandin I2 (prostacyclin)
PUFA
PolyUnsaturated Fatty Acid
TNFa
Tumor Necrosis Factor
TxA2
ThromboXane A2
PLA2
PhosphoLipase A2 (enzyme)
Substrate: cell membrane phospholipids
Fxn: liberates AA
FLAP
5-LO activating protein (enzyme)
Substrate: 5-LO accessory protein
Fxn: converts AA + O2 -> LTA4
LTA4 hydrolase
Enzyme that converts LTA4 -> LTB4
LTC4 synthase
Enzyme that converts LTA4 -> LTC4
Peptidases
Enzymes that convert LTC4 -> LTD4 -> LTE4
LTA4
Fxn: biosynthetic intermediate
involved w/asthma
LTB4
Receptor: BLT1,2
Fxn: augments neutrophil EC adhesion, POTENT neutrophil chemotaxis and degranulation, eosinophil chemotaxis
involved w/asthma
main LT made by neutrophils
LTC4 = LTD4
Receptor: CysLT2
Fxn: bronchoconstriction, mucus secretion
involved w/asthma
LTD4 > LTC4
Receptor: CysLT1
Fxn: bronchoconstriction, mucus secretion
involved w/asthma
LTE4
Receptor: CysLT
Fxn: less active metabolite
involved w/asthma
Histamine
Synthesized and stored pre-formed (latent) in granules of mast cells and basophils
Substances in granules (ex- heparin) form complexes w/histamine to keep inactive until released
Increases capillary permeability to leukocytes/plasma proteins to attack pathogens in tissue
Histamine synthesis/inactivation for storage
Dietary histidine -(decarboxylase) -> CO2 + Histamine
- Diamine oxidase: imidazole aldehyde (inactive)
- Histamine N-methyl transferase: N-methyl histamine (inactive)
Histamine v Prostaglandins/Thromboxane/Leukotrienes
Synth/Storage: Histamine made all the time, RELEASED on demand; P/T/L precursors stored, MADE on demand
Lifespan: both short, Histamine inactivated enzymatically, P/T/L inactivated enzymatically OR spontaneously
Physiology: both local (not systemic)
Pathology: both due to unwarranted SYSTEMIC exposure
H1 receptor
tissue-specific GPCR for histamine
Nasal/bronchial- mucus secretion
Bronchial/intestinal smooth muscle- constriction
Sensory nerves- pain
H2 receptor
tissue-specific GPCR for histamine
Stomach- gastric acid secretion
H1 and H2 receptors
tissue-specific GPCRs for histamine
Heart- HR
Vessels- peripheral resistance
Skin capillary blood vessels- dilation, permeability, pain sensitization
COX-1
helps maintain normal cell fxn
constitutive in platelets, ECs, fibroblasts, smooth muscle, monocytes, macrophages, mast cells, basophils, neutrophils (less)
COX-2
inducible in ECs, fibroblasts, smooth muscle, monocytes, macrophages, mast cells, basophils, neutrophils (less)
Leukotriene synthesis
5-LO arm of AA cascade, lipid/peptide hybrid
1. PLA2: AA released from membrane
2. 5-LO, Ca2+ + FLAP: O2 inserted into AA -> LTA4
3. 2 pathways
LTA4 hydrolase: adds H2O -> LTB4
LTC4 synthase: adds GSH -> LTC4
peptidases: LTC4 -> LTD4 and LTE4
Leukotriene breakdown
spontaneous hydrolysis into inactive products
helps prevent LT excess
Self-reliant chemotaxis (neutrophils)
neutrophils encounter threat -> inflammation favors eicosanoid synth in neutrophils -> 5-LO makes lipid mediators (LTs) -> LTB4 (augments EC neutrophil adhesion, potent chemotactic agent for neutrophils)
pyogenic infection
characterized by neutrophil-rich pus
Mediators of inflammation
Made by phagocytes and granulocytes after they engulf the threat and release ROS and proteases
Send autocrine and paracrine signals telling leukocytes to engage/neutralize threat
Tells epithelial and mesenchymal cells to adapt/migrate/perish
Limit damage to host due to infammation
Asthma elements
- airway inflmamation (Th2 lymphocytic response-> secrete IL-4/5/13, eotaxin (chemokine), TNFa, LTB4, tryptase (mast cell))
- airway hyper-responsiveness (coughing triggered more easily than normal person
Asthmatic airway
bronchiolar smooth muscle constriction
inflammation
mucus discharge
pulmonary edema
Hypersensitivity
- Sensitization: allergen exposure-> IgE abs
- IgE abs bind mast cell receptors
- Re-exposure: allergen binds IgE-> CROSS LINKS RECEPTORS
- Cross-linking triggers DEGRANULATION-> histamine release-> activate eicosanoid synth (LTC4/D4, PGD2)
- Mediators + histamine -> redness, local edema, pain, itching (symptoms of allergic immediate hypersensitivity)
Histamine hypersensitivity
Originates from H1 receptors on vessel wall (esp. venous side)
Causes vasodilation and inc permeability
Causes pain (mechanical pressure on peripheral nerves due to tissue swelling)
Mast cell/histamine location
Concentrated in areas of the body most vulnerable to antigen/pathogen exposure Nasal passage Trachea, bronchi, lungs Stomach, intestines Skin
allergy treatment
inhibit RECEPTORS specific to area w/symptoms
inhibiting synthesizing enzyme risks having a shortage of histamine in other important areas of the body
Circulating inflammatory cells
Neutrophils: 40-60% lymphocytes: 20-40% Monocytes: 2-8% Eosinophils: 1-4% Basophils: .5-1%
Gout
Failure to excrete uric acid at an appropriate rate
Treat inflammation AND metabolism (synth) of uric acid
Gout mechanism
- urate crystals in joint -> sterile inflammation
- MP/monocyte engulfs crystals -> releases IL-1b (inflam) and IL-8 (chemotactic)
- IL-1b -> EC adhesion molecules and MP/monocyte/conn tissue/EC COX-2
- IL-1b + IL-8 -> neutrophil recruitment
- MP/monocytes/neutrophils/conn tissue -(COX-2)-> local inc lipid inflam mediators (dil/perm) -> redness/swelling/heat/pain
IL-1 beta (gout)
Binds membrane receptor
Releases substrate (AA) and activates enzyme (COX-2)
Induces adhesion molecules in EC (inc neutrophil margination)
PGE2 and PGI2 (gout)
PGI2: vasodilation
PGE2: permeability
Leaky vessels-> plasma extravasion-> local edema
(redness, heat, swelling, pain)
IL-8 (gout)
Chemotactic factor initiating movement of neutrophils from blood -> leaky vessels -> tissue w/activated MP/monocyte
Neutrophil adaptability
Normal: reside in blood
Pathology: EXIT blood, enter tissue site, 2 fxns
1. granulocyte: discharge degradation mediators
2. phagocyte: engulf and digest
Inflammation (gout)
fails to eliminate uric acid crystals
ineffective proteases and lysosomal enzymes (phagocyte)
ineffective oxidative bursts (granulocyte)
May aggravate: uric acid ppts at acidic pH
Bacteria vs gout
Stimulus: LPS/bact/fungus v Uric acid crystals
Cytokines: TNFa/IL-1/IFNg v IL-1b/IL-8
Involvement: Systemic v Local
Inflammation (bacteria)
- Bact/LPS -> inflam
- MPs/monocytes -> inflam (TNFa/IL-1/IFNg) and chemotactic cytokines
- LPS/TNFa/IL-1 induce EC adhesion molecules and MP/mono/conn tiss/EC COX-2
- Neutrophil recruitment by cytokines/inflam med.s
Chlorox
mimics hypochlorous acid in our phagocytic cells (bacteriocidal oxidative burst)
Septis
Systemic inflammation due to circulating cytokines
Damage caused by COX-2, adhesion molecules, and myeloid activation
Systemic vasodilation and plasma extravasion -> BP drop
Can occur if microbe enters bloodstream (body mounts immune response towards entire bloodstream)
Can lead to septic shock (low BP), multi-organ failure, and death
Anaphylaxis
- Exposure (resp tract to allergen)
- Activation: mast cells (bound IgE receptor)
- Degranulation: mast cells-> histamine
- Respiratory excess: airway constr, impaired breathing
- Vasculature excess: hypotension (10 min: histamine inc, 30-60 min: normal)
- Urine: histamine and N-methylhistamine
BONUS: mast cells make LTC4, LTD4, PGD2 -> inc perm, dec vasc periph resist, hypotension, mucus, bronchoconstr
TREAT: epinephrine (vasoconstr/bronchodil) and agents to counter histamine/LTs/eicosanoids
Self-Nonself Model
Each lymphocyte expresses a single surface receptor specific for a foreign Ag
Receptor signaling initiates imm resp
Self-reactive lymphocytes deleted EARLY in life
Danger Signals
Stimulate DC maturation in lymph node
- microbe infection
- necrosis/stress products
- immunostimulators (heparan sulfate)
- inflam cytokines
- vessel rupture/chemotaxis
Dendritic cells vs macrophages
DCs have much higher expression of costimulatory molecules
Macrophages don’t have enough costim expression to activate naive T cells
ONLY DCs can activate naive T cells
M. tuberculosis T cells
Th1
prevents fusion of phagosome and lysosome, grows slowly, more common in crowded/hot/humid conditions b/c transmitted by droplets
Asthma T cells
Th2
Chemokine receptor and example
GPCR, IL-8
most potent inducer of TNFa/IL-1
LPS/endotoxin
keloid scar
TGFb -> myofibroblasts -> fibrosis -> keloid scar
fibroblasts vs myofibroblasts
myofibroblasts have more actin, make more conn tissue
Innate cytokines
TNF/IL-1, IL-6
IL-8, IL-10, IL-12
IFN (a/b/g)
IL-15, IL-18
Adaptive cytokines
IL-2- T cell prolif/diff (Tc/Th/Treg) IL-4- IgE switch IL-5- eosinophils, IgA IFNg- M1 IL-17- inflam TGFb- Treg, inh T cells
T-bet
transcription factor for Th1 (intracellular bacteria)
GATA-3
transcription factor for Th2 (helminths)
RORgT
transcription factor for Th17 (extracellular bacteria/fungi)
FOXP3
transcription factor for Treg cells
IL-2R
naive T cell: low affinity, beta-gamma-c
activated T cell: high affinity, alpha-beta-gamma-c
stimulates IL-2R expression on activated T cells (after a few hrs)
decreases w/dec in antigen
IL-12 knockout mice
can’t control m. tuberculosis infection because no Th1 (adaptive/cell-mediated immunity)
die late
IFNg knockout mice
die early because macrophages (M1) aren’t activated-> no phagocytosis/inflammation (innate immunity)
granulomas
created by M2 macrophages
Type I cytokine receptor
hemopoietin Jak STAT IL-2/3/4/5/6/7/9/11/12/13/15/21 IL-2Rbgc/IL-2Rabgc G/GM-CSF
cytokine synergy
synergistic EFFECTS from receptors, but each cytokine has different receptor (receptors don’t actually work together)
Type II cytokine receptor
Jak STAT
IFNa/b/g
IL-10
TNF receptor
TRAF I: TNFrp75 II: TNFrp55 TNFa TNFb/lymphotoxin LTs
IL-1 receptor
IRAK
IL-1/18
chemokine receptor
(IL-8)
GPCR
Positive T cell selection
Thymus: method of maturation of MHC-restricted T cells
positively select only T cells w/WEAK recognition of MHC I/II (one or the other)
Failure (no MHC recognition) -> apoptosis
Too much strength could lead to hypersensitive T cells that respond to self
Negative T cell selection
Thymus: method of maturation of MHC-restricted T cells
negatively select against T cells w/STRONG recognition of MHC I/II
Too much strength could lead to hypersensitive T cells that respond to self
T cell co-receptors
CD4 and CD8
T cell activation
- skin: langerhans cells uptake antigen
- LH cells w/Ag enter lymphatic system
- LH cells enter lymph node (afferent vessel) -> become DCs that have B7
- LN: B7 DCs activate naive T cells (parafollicular cortex)
afferent lymphatic vessel
where activated DCs enter LN to deliver Ag to T cells
parafollicular cortex
T cell zone of LN
where activated DC (w/B7) activate T cells
CD3 proteins
gamma, delta, epsilon, zeta
needed for T cell activation with TCR and CD4/8
cytoplasmic tail is long enough to signal
T cell activation proteins
Costimulation**** TCR + Ag CD28 + B7 (CD80/86) CD3 CD4 or 8 + MHC II or I CD40L + CD40
T cell stimulation process
1st interaction w/APC: produce ONLY IL-2, NOT committed to Th1/2
2nd interaction w/APC: produce OTHER CK’s, commit to Th1/2, effector cell HOMING, memory cells stay in LN
Th1-> IFNg, TNFb, IL-2
Th2-> IL-4, IL-5, IL-10
T cell homing
Occurs with RE-stimulation (2nd interaction w/APC)
T cells return to part of body from which APC’s (tissue macrophages) came
effector T cells move from LN -> tissue
memory T cells remain in LN
Th1/2 polarization
takes about 2 days
memory cells remaining in LNs help skip this delay in subsequent infections
Th1 differentiation results from…
presence of activated macrophages/DCs and IL-12
default unless no IL-12
Th2 differentiation results from…
presence of IL-4
ABSENCE of IL-12
CD40-CD40L
T cell activation (by APC)-> express CD40L
CD40L -> binds CD40 (on APC) and increases MHC/B7 (on APC), helps w/B cell activation
CD40 -> inc APC potency
Ab activation
need Ag binding
Exception: mast cells w/IgE (bind IgE’s Fc)
Cross-presentation
LN: DCs present intracell microbes on MHC I and extracell microbes on MHC I and II
CD8 T cells recognize MHC I, CD4 T cells recognize MHC II
CD4 T cells and DCs stimulate CD8 T cells
CD8 T cells proliferate -> leave LN
Granzyme
enters target cells through
- receptor-med endocytosis
- membrane holes created by perforin
gd T cells
gd TCR (not alpha/beta) + CD3 (no CD4/8) Location: intestine, uterus, tongue (epithelium) Function: 1st line of defense, regulation (make CK's), link inn/adap Adaptive: TCR gene rearrangement, memory Innate: PRRs do NOT need APCs (or MHC)
NKT cells
T and NK CDs
recognize lipids and glycolipids (w/CD1d)
helps against Tb
secrete IFNg and IL-4
deficient NKT cells
autoimmunity
cancer
asthma progression
somatic recombination
gene rearrangement in non-dividing/somatic cell (ex- immune cell) to create T/B cell diversity
lots of mutations due to lack of DNA repair mechanisms
combinatorial diversity
TCR a/b chains and BCR (Ig) H/L chains
beta/Heavy: VDJ recombination
alpha/Light: VJ recombination
Followed by transcription, mRNA splicing, translation
junctional diversity
increases BCR and TCR diversity
due to random NT removal/addition
exonuclease and terminal deoxyribonucleotidyl transferase (TdT)
almost unlimited
somatic hypermutation
Only in B cells, after Ag exposure/memory re-stim
Point mutations in heavy chain and variable region due to C->U (deamination) repair (U replaced with incorrect NT)
May lead to affinity maturation (inc ab affinity for its epitope)
Followed by selection of high-aff B cells by Ag-presenting follicular DCs (in lymphoid follicle germinal centers)
active immunity
acquired by T cells after disease
Double-negative T cell
precursor cells from bone marrow w/o TCRs or CD4/8
reside in subcapsular cortex region (thymus)
Double-positive T cell
begin TCR gene rearrangement
both coreceptors expressed
reside in deep cortex (thymus)
Single-positive T cell
inc TCR expression
lose either CD4 or 8 (whichever receptor binds self too strongly)
reside in medulla (thymus)
T helper mechanism
- Macrophage engulfs microbe, breaks down, presents to Th on MHC II
- CD4 Th w/proper specificity undergoes clonal expansion -> effector cells and memory cells
- T effector cells interact w/B cells (B cells w/proper specificity neutralize Ag and undergo clonal expansion-> plasma cells and memory cells)
- Plasma cells secrete Igs to block Ag
complementarity determining regions
regions where light chains are complementary to heavy chains
in variable region of light chains and heavy chains
Kappa light chain genes
L/V: 35 (but maybe 300) (2 exons: L and V)
no D region
J: 5 (b/w V and C)
Constant: 1
Rearrangement: V+J
Lambda light chain genes
L/V: 30 (2 exons: L and V)
no D region
J: 4 (b/w V and C)
Constant: 4
Rearrangement: V+J
complementarity determining regions
regions where light chains are complementary to heavy chains
in variable region of light chains and heavy chains
CDR3 is most variable and most IMPORTANT for Ag recognition
Combinatorial diversity
increases BCR and TCR diversity
due to somatic recombination
V(D)J recombinase
limited by available V/D/J gene segments
V(D)J recombinase
COLLECTION of enzymes in immature B/T cells
sloppy somatic recombination of VDJ gene segments in B/TCR
Recombinase-activating gene (RAG)-1 and RAG-2
Exonuclease
Ligase
RAG-1 and 2
bind recombination switch sequence spacer (12-23 bp b/w heptamer and nonamer)
TdT
randomly adds NTs to V/D/J gene segments at site of V(D)J gene recombination
contributes to hypervariability of CDR3
CDR3
most variable CDR in V region
most IMPORTANT for Ag recognition by B/T cells
human leukocyte antigens
MHC proteins
each molecule has 1 peptide-binding cleft
determine graft acceptance between individuals
highly polymorphic (only same if monozygotic twins)
express paternal/maternal equally (codominant)
3 million bps on Chr 6
MHC I
binds peptides (8-10 AAs) in the CYTOSOL recognized by CD8 CTLs presents peptide fragment epitope at antigen binding cleft Domains: a1, a2, a3, b2m a1/a2 (cleft) vary b/w people a3 invariant, binds CD8 beta2microglobin maintains conformation
MHC II
binds peptides (13-25 AAs) from w/in VESICLES
recognized by CD4 Thelpers
presents peptide fragment epitope at antigen binding cleft
Domains: a1, a2, b1, b2
a1/b1 (cleft) vary b/w people
b2 binds CD4
MHC I genes
HLA-A, B, C (D/E/F not important)
inherit one set from each parent -> any cell can have 6 diff MHC I molecules
MHC II genes
HLA-DP, DQ, DR (DM/DO not important)
alpha/beta chain = polymorphic -> any cell can have 10-20 diff MHC II molecules
autografts (autologous)
tissue grafted from one place to another in same person
ex- skin
isografts (syngeneic)
tissue transplants b/w genetically identical people
allogenic grafts
tissue grafts b/w genetically different members of SAME species
ex- kidney transplants
xenogenic grafts
tissue grafts between members of DIFF SPECIES
ex- pig heart valves for humans
Transplant Test
ABO blood typing compability
HLA typing
preformed Abs
Crossmatching
hyperacute graft rejection
caused by ABO blood type incompabilities
HLA typing
focuses on HLA-A, B, DR
more HLA matches = better graft survival
haplotype
set of alleles of linked genes one 1 parental chromosome
determine different antigens, but inherited as unit
minimized chance of crossing over
never identical unless monozygotic twins
new haplotype can occur within same individual if recombination takes place
haplotype matching
need same genes AND same sequence
B7-1/2
co-stimulatory molecule (CD80/86) expressed by activated APCs with MHC II
Ag binding activates APCs and increases B7 expression
binds CD28 on T cell to activate (w/MHC II)
Necrosis
passive, catabolic cell death in response to external toxic factors
induces caspase cascade
characterized by swelling and rupture of cell membrane (lysis), which may cause inflammation or harm other cells
Inflammation
physiological: eliminates initial cause removes necrotic cells initiates repair pathophysiological: injury bystander normal tissue normally self-limited, can become chronic
Humoral response to danger signals
activation of complement
activated immune cells: chemokines, leukotrienes/prostaglandins, ROI, NO
necrotic danger signals
HMGBI
Uric acid
Heat Shock Proteins
acute inflammation process
- Detect damage: vascular coagulation, PRRs recognize pathogens/cell injury
- Leukocyte Recruitment and Stimuli: C5a, PRR-> EC adhesion molecules and plasma exudation (neutrophils first)
- Resolution: Microorganisms/necrotic tissues eliminated-> apoptotic neutrophils phagocytized by MPs (scavenger receptors)
- Wound healing: angiogenesis, re-epitheliziation, collagen deposition, MP-(TGFb)-> fibroblasts
Uric acid
necrotic danger signal
activates NFkB
HSPs
necrotic danger signal
activates NFkB and release of pro-inflam CK’s (TNFa/IL-1b)
HMGB1
necrotic danger signal received by DCs High Motility Group Box 1 passively-released protein during necrosis activates NFkB Receptor: RAGE
Uric acid
necrotic danger signal received by DCs
activates NFkB
HSPs
necrotic danger signal received by DCs
activates NFkB and release of pro-inflam CK’s (TNFa/IL-1b)
RAGE
DC Receptor of Advanced Glycation End Products
receptor for high motility group box 1 (HMGB-1, necrotic danger signal)
chronic inflammation
cancer diabetes cardiovascular neurological disease autoimmunity arthritis pulmonary disease alzheimer's disease
artherosclerosis
- monocytes recruited by activated ECs -> MPs
- TLRs recognize microbes -> activate MPs (foam cells filled w/lipids)
- Pro-inflam CK’s, ROI, NO, etc.
- Inflammation and tissue damage
- MP’s accumulate lipids-> become foam cells
scavenger receptors
no feedback mechanism(/refractory period) (unlike normal receptors that have refractory periods)
statins
break down lipid foam cells (possible correlation w/Alezheimers)
Apoptosis molecular triggers
DNA damage CK starvation hypoxia temperature death receptors
Apoptosis molecular regulators
Maintain equilibrium b/w pro- and anti-apoptotic signals Death domain factors cytochrome c p53 Bcl-2 family Myc/oncogenes
Apoptosis molecular executioners
caspases
apoptosis-inducing DNA damage ex
DNA damage: keratinocyte exposed to UV
Cytokine: cells w/o CKs
Caspases
cysteine proteases
orchestrate morphologic changes
destroy key components of cellular infrastructure-> initiate apoptosis
Apoptosis molecular mechanisms
- Intrinsic (mitochon), Extrinsic (Fas, TNFr), CTLs (granzyme), Injury (toxins, free radicals)
- pro-apoptotic molecules (ex- cytochrome c)
- executioner caspases -> endonucleases and cytoskeleton breakdown
- phagocytic cell receptor ligands (cytoplasmic bud-> apoptotic body-> phagocytosis)
Fas
CD95
expressed by every cell
only activated lymphocytes express Fas ligand
Intrinsic Mitochondrial pathway
TRIGGERS: 1. Bcl-2 (BAK/BAX) 2. Ca 3. Free radicals REGULATORS: 1. Cytochrome c 2. Smac/Diablo 3. Apoptosis-inducing factor 4. Endonuclease G EXECUTIONERS: (not immediatly activated) - Caspase 9 - Caspase 3 - Apaf-1
Caspase activation
executioner phase of apoptosis
“point of no return”
Extrinsic Apoptotic pathway
- Fas ligand (TNF ligand family) binds Fas (TNFr family)
- FADD activation
- death effector domain activation
- procaspase-8, 10 -> Caspase 8, 10 -> cascade
- mitochondrial damage, membrane changes, proteolysis, nuc condensation/DNA fragmentation
- APOPTOSIS
Autoimmune Lymphoproliferative Syndrome
ALPS patients: heterozygous mutations in Fas gene
early life: chronic adenopathy/splenomegaly
chronic persistence/activation of both T cells -> B cell maturation -> Ab secretion
defective Fas-med apoptosis-> extended survival of lymphocytes -> possible malignant transformation
T cells normal in beginning, but don’t die-> LN swelling
B cell selection
- Negative selection against B cells w/HIGH affinity for self antigen
- Receptor editing: self Ag recognition reactivates Ig gene recomb-> new light chain expressed (not specific for self antigen)
Negative selection
first part of B cell selection
gets rid of B cells w/HIGH affinity for self antigen (apoptosis)
receptor editing
second part of B cell selection
when BCR recognizes self antigen, it reactivates Ig gene recomb and creates a new light chain NOT specific for self antigen
B cell activation (by Ag)
- Ag recognition by naive IgM+/D+ B cell
- B cell activation by helper T cells and other stimuli
- B lymphocyte activation/Clonal Expansion
- Differentiation to effector cells (ab-secreting plasma cells, ab-expressing B cells, high-affinity Ig-expressing B cell)
Effector fxns: Ab secretion, isotype switching, affinity maturation, memory B cell
IgD
membrane bound, high in new born babies
B cell can make ____ plasma cells which make ____ abs per day
4000
10^12
B cell Ag recognition (occurs where, initiates what)
occurs in LN
Ag does not require processing
activates B cells
Initiates…
1. B cell proliferation (enter cell cycle-> mitosis)
2. Inc expression of costim (MHC II, B7) and CK receptors (Thelper cells)
3. Low levels of IgM secretion
B cell gene mutations
V/D/J gene segments have a mutation every 1000 bases (normal DNA has 1 every 10^8)
called somatic hypermutation
results in affinity maturation
B cell proliferation
must constantly be RE-stimulated by binding to their cognate antigens
higher affinity BCR (diff ones due to somatic hypermutation) means they get Ag first-> stimulated first/more easily/more
Result: more B cells w/high affinity BCR for Ag
C3d
protease-modified version of C3b (from innate complement system)
C3b stays bound to microbe-> persists and modified to C3d bound to microbe
recognized and bound by CR2 (adaptive immunity) on B cell
CR2
adaptive immunity receptor
helps B cells recognize microbes early in infection when Ag is low
binds C3d (bound to microbe) and increases Ag sensitivity of BCR 100x (helps activate B cell)
B cell migration
B cells activated in the lymphoid follicle (B cell zone)
Migrate to central zone to interact w/T follicular helper cells in parafollicular cortex (T cell zone)
parafollicular cortex
T cell zone of LN
lymphoid follicle
B cell zone of LN
T follicular helper (Tfh) cells
interact with B cells in central zone of LN
B cells vs DCs (as APCs)
B cells: activated by Ag/BCR or rec-med endocytosis-> Ag processing-> presented on MHC II to T helper cell
DCs: rec-med endocytosis-> Ag processing-> presented on MHC I to CTLs
Hyper IgM Syndrome
caused by defect in ability to switch from production of IgM to IgG/A/E
defect is either in CD40L (X-linked) or CD40
CD40L defect
results in Hyper IgM syndrome
X-linked-> ONLY MALES AFFECTED
CD40 defect
results in Hyper IgM syndrome
affects males and females equally
B cell Activation (by T cells)
- cytokines + direct contact b/w B cell and Th cell
- activated Th cells have CD40L that binds CD40 on B cells -> costimulation
- BCR cross-linking-> B cell activation
- B cell prolif, initial ab production (IgM w/some IgD), germ center rxn
tiger licks baby
- mom licks Ags off cub
- Ags picked up by IgA in intestine
- Ag-specific IgA and Ags secreted in breast milk
- cub drinks milk
- cub develops OWN IgA abs against Ags
- establishment of Humoral Imm Sys
BREAST FEEDING ESTABLISHES IMM SYS
final step in B cell maturation
PLASMA CELL
or
MEMORY CELL
plasma cell lifespan
~5 days (make 2000 abs/sec)
plasma cell location
spleen
bone marrow
memory cell
depends on CD40-CD40L interaction
NO MEMORY WITHOUT B CELL ACTIVATION BY T HELPER CELLS
Thymus-dependent antigens
the only ones T cells help with proteins isotype switching affinity maturation MEMORY B CELLS
Thymus-independent antigens
POLYMERIC proteins, carbs, lipids, nucleic acids that persist for long periods of time (resistant to degradation)
IgM, some IgG (no switching)
no affinity maturation
only some Ags produce memory
can’t bind MHC II (no T helper cells)
cross-link many BCRs (polyclonal)-> activate B cells-> prolif/diff
TI-1 and TI-2 Ags
TI-1 Ags
Thymus-independent
polyclonal activators of B cells
NON-SPECIFIC activation of multiple B cells at a time
TI-2 Ags
Thymus-independent
not polyclonal
repeating epitopes for cross-linking BCRs
SPECIFIC activation of clones of ONE B cell
Abs have relatively low affinity for TI-2 Ags
B cell activation (by TI-1 Ags)
Signal 1: BCR binds Ag (ex- LPS)
Signal 2: TLR binds Ag
Multiple B cells initiated (polyclonal)
B cell activation (by TI-2 Ags)
Signal 1: BCR binds Ag
Signal 2: BCR CLUSTERING/cross-linking
Ags are strongest inducer of COMPLEMENT (b/c multi-epitopal)
Abs produced: mostly IgM (important for encapsulated bacteria)
Relatively low Ab affinity for TI-2 Ags
encapsulated bacteria
major mechanism of host defense is Ab-mediated Immunity (IgM**)
bacterial cell wall polysaccharides = TI Ags
Activate B cells right away without having to wait for T cell activation
Ab-med immunity deficiency (susceptibility to…)
congenital or acquired deficient Ab-mediated imm response
very susceptible to infections w/encapsulated bacteria
capsule=TI Ag that would usually induce complement but have to wait for T cell activation, giving bacteria a chance to flourish
latent infection
immune response controls but does not eliminate the infection
microbe is good at “hiding”
usually slow growing, causes cell lysis
ex- HIV, Tb
cause of tissue injury/disease
usually due to host immune response to microbe (rather than due to microbe itself)
SCID
no adaptive imm (B/T cells)
innate immunity suppresses initial infection, but eventually microbes cannot be contained
extracellular bacteria pathological mechanisms
- inflammation (tissue destruction)
2. toxin production (endo/exo)
Endotoxins
components of bacterial cell walls
only released upon CELL DEATH
ex- LPS (potent activator of MPs, DCs, and ECs)
Exotoxins
secreted by bacteria when they’re ALIVE
many are cytotoxic (ex- diphtheria/cholera/tetanus toxin)
some interfere w/normal cellular fxn (but don’t kill cell)
some stimulate CK production-> cause disease
diphtheria toxin
exotoxin from extracellular bacteria
Fxn: shuts down protein synthesis in infected cells
cholera toxin
exotoxin from extracellular bacteria
Fxn: interferes w/ ion and water transport
tetanus toxin
exotoxin from extracellular bacteria
Fxn: inhibits neuromuscular transmission
complement
PGN (G+) and LPS (G-) -> alternative Mannose -> lectin MAC -> bact lysis (ex- Neisseria) C3a + C5a -> leukocyte chemotaxis/activ C3b -> MAC or opsonization (for phagocytosis) Result: enhanced phagocytosis
acute phase proteins
mannose-binding lectin
C-reactive protein
Phagocytes
neutrophils and macrophages
Recognize extracell bact: MANNOSE and SCAVENGER receptors (which then promote phagocytosis)
Recognize opsonized bact: Fc (Abs) and COMPLEMENT receptors (which then promote phagocytosis AND activation)
microbial products activate them via TLRs
Activation-> phagocytes secrete CK’s-> leukocyte infiltration to infection site (inflammation)-> leukocytes ingest/destroy bacteria
ROI effects
- DNA damage
- Lipid peroxidation
- AA oxidation
- Enzyme inactivation by co-factor oxidation
Innate immune evasion by extracellular bacteria
inhibit complement activation (many bacteria) resist phagocytosis (pneumococcus, Neisseria meningitidis) scavenge ROI (catalase+ staphylococci)
extracellular bacteria that resist phagocytosis (ex)
pneumococcus, Neisseria meningitidis
extracellular bacteria that scavenge ROS (ex)
catalase+ staphylococci
peroxynitrite (ONOO-)
causes apoptosis (mitochondria: AIF, or cit C-> caspase activation) or necrosis (lipid peroxidation, protein oxidation, protein nitration, inactivation of enzymes)
innate immune response against extracellular bact
- complement (opsonization-> phagocytosis and MAC-> lysis)
- inflammation
- phagocytosis (neutrophils and macrophages)
adaptive immune response against extracellular bact
Antibodies…
- neutralize microbes/toxins
- opsonize microbes for phagocytosis
- help NK cells w/Ab-dep cytotoxicity
- complement (lysis, opsonization/phagocytosis, inflammation)
Ab neutralization
prevents bacteria/toxin from binding cells/infecting new cells
utilized by vaccines-> induce Th2 response-> IgE production
inc vaccines may correlate w/inc allergies in modern humans (body prefers to use IgG, not IgE
Ab-mediated phagocytosis
- IgG opsonizes microbe
- IgG binds phagocyte Fc receptor (FcgR)
- phagocyte activation
- phagocytosis
- killing
Fc receptors
FcgRI: IgG
FceRI: IgE
FcaRI: IgA
none for IgM (pentamer-> can’t bind)
phagocyte Fc receptors
FcgRI: IgG
FcaRI: IgA
bind Ab-Ag COMPLEXES ONLY
FceRI
binds FREE IGE (not in complex)
On mast cells, eosinophils, basophils
binding-> granule secretion
extremely high binding strength means blood levels of IgE are never very high b/c immediately taken up by mast cells
adaptive immune evasion by extracell bacteria
antigenic variation (Neisseria gonorrhoeae, E coli, salmonella typhimurium) inhibit complement activation (many bacteria) resist phagocytosis (pneumococcus, Neisseria meningitidis) scavenge ROI (catalase+ staphylococci)
antigenic variation
helps extracellular bacteria evade adaptive immunity
bacterium/virus alters surface proteins to evade host immune response
ex- Neisseria gonorrhoeae, E coli, salmonella typhimurium
complement inhibition
helps many extracellular bacteria evade innate and adaptive immunity
bacterial capsules, C3 convertase decay, blocked MAC formation (vitronectin)
vitronectin
host protein used by bacteria to block MAC formation and inhibit complement
phagocytosis resistance
helps extracellular bacteria evade innate and adaptive immunity
interfere w/complement activation and/or deposition at bacterial surface
inject anti-phagocytic effectors into cell
ex- pneumococcus, Neisseria meningitidis
scavenge ROI
helps extracellular bacteria evade innate and adaptive immunity
catalase-pos pathogens deactivate peroxide radicals-> survive unharmed WITHIN host
ex- catalase-pos staphylococcus
pyrogens
cause fever (INNATE RESPONSE) Endotoxin/LPS (G-) LTA (G+) viruses yeast molds environment (packing materials)
fever
innate response caused mostly by IL-1, but also IL-6, and least by TNFa
manifestation of CK signals to hypothalamus
bacterial pneumonia
fever => infection
localized crackles => bacteria (crackles everywhere=> virus)
otherwise healthy => ACUTE
Labs: lung x-ray, sputum sample, complement testing (C3 levels)
Do NOT wait for lab results, START W/BROAD ABX b/c infection could lead to septic shock
Immunological Tolerance
Specific unresponsiveness to an Ag (ex- self-tolerance)
Breakdown-> autoimmunity
Imperfect negative selection of self-reactive T lymphocytes-> low level of auto-reactivity (crucial for normal imm fxn)
Remission
Occurs because Ags are removed from imm sys but NOT from the BODY