B1 Review

Saturday, December 13, 2014 at 9:45 AM

Innate vs Adaptive Immunity

Innate
1st response - always active
PAMPs recognized by PRRs
Inbuilt/Innate (present from birth)
Immediate/Rapid response
Non-sepcific
Limited Diversity
NO Memory
cells involved:
Phagocytes (Neutrophils, Monocytes, Macrophages)
Granulocytes (Mast cells, Eosinophils, Basophils)
NK cells
Adaptive Immunity
2nd response - needs to be induced
done by CD4+ Th cells
Antigens are recognized by BCRs and TCRs and antibodies
Induced/Activated
Delayed Response
Specific
Diverse
Tolerance of self
Memory
cells involved:
Lymphocytes
T cells and B cells ONLY
NK cells are lymphocytes, but they are part of INNATE immunity
Immune Response
Humoral
1) APCs present Ag to CD4+ Th cells
2) Th cells help B cells to differentiate into Plasma cells
3) Plasma cells produce antibodies
4) Abs bind to Ag directly, or a product of the pathogen
Abs provide defence against EXTRACELLULAR pathogens
Cell-Mediated
CD8+ CTLs kill our own cells that are infected with INTRACELLULAR pathogen
they do NOT kill the pathogen directly, but rather the cell hosting the pathogen
CD8+ cells only become CTLs with help from CD4+ Th1 cells
NK cells already have cytotoxic granules, so they dont need to be activated
they participiate in ADCC (antibody dependent cell cytotoxicity)
Lymph Nodes
B cell zone = Primary Follicle, or Cortex
T cell zone = Paracortex
Germinal Center = houses ACTIVATED T and B cells
only form after 1st T-B cell help and SHM
Spleen
B cell zone = Primary Follicle, or Cortex
T cell zone = Periarteriolar Lymphoid Sheath (PALS)
Mucosal Associated Lymphoid Tissues (MALT)
Peyers Patches = lymphoid tissue in the gut, covered by mucosal epithelium
M cells allow transcytosis of molecules, so dendritic cells can sample the environment
Cells of the immune system
Myeloid Lineage = includes the cells of the INNATE Immune system, with exception of NK cells (which are Lymphoid)
Eosinophils, Basophils, Neutrophils, Monocytes (which become Dendritic cells or Macrophages), Mast cells
Lymphoid Lineage = cells of the ADAPTIVE Immune System
T cells, B cells, NK cells
NK cells are actually Innate
Primary (Central) Lymphoid Organs/Tissue
Bone Marrow and Thymus
Secondary (Peripheral) Lymphoid Organs/Tissue
Spleen, Lymph Nodes, MALT
Leukocytes (WBCs)
Granulocytes
Neutrophils = most common WBC, kill via phagocytosis
dominates the immediate response
Mast cells are Tissue-Resident cells that deal with Parasites
Eosinophils and Basophils also deal with parasites
Agranulocytes
Monocytes (which become DCs or Macrophages) and Lymphocytes
Bacterial Structure
Cocci = ball-shaped
Staphylococci
Bacilli = rod-shaped
E. Coli
Spirochetes
Borrelia (Lyme Disease)
Pleomorphic = no identifiable shape
Vibrio Cholera
Gram Staining
Gram Positive (Gram+)
cell wall has THICK layer of Peptidoglycan
Purple/Blue staining color
Staph Aureus, Streptococcus, Clostridium, Listeria
Gram Negative (Gram-)
cell wall has THIN layer of Peptidoglycan, encased in 2 layers of cell membrane
outer cell membrane contains Lipopolysaccharide (Endotoxin)
Pink staining color
E. Coli, H. Influenzae, N. Meningitides, N. Gonorrhea
Acid-Fast Stain
stains bacteria that have a high LIPID and WAX content in cell wall, and Mycolic Acid
also stains bacteria with NO cell wall
Mycobacterium, Nocardia, and Mycoplasma
External Structures
Flagella = motility
Axial Filaments or Endo-flagella
flagella contained between the 2 plasma membranes (Gram-)
ONLY found in Spirochetes
Pilus and Fimbria
Sex Pili (F Pili) = adhesion and transport of bacterial proteins or DNA Conjugation
Capsules-Glycocalyx slime layer
protection against killing by immune cells, and Ab penetration
resistance to phagocytosis
Biofilms
layer of protection over an entire population of bacteria
Internal Structures
Genetic Material
plasmids, circular chromosomes
Chemoattactants
initial AA in all prokaryotic proteins is Formylated Methionine
Endospores
survival forms, highle resistant and protective
Secretion Systems
Type I secretion = standard secretory protein that releases molecule into the ECF
Type III secretion = needle that extrudes outside the cell, allowing injection of material directly into target
both Types are used by Gram NEGATIVE
Gram+ thick cell wall prevents secretion
Bacterial Products
Exoenzymes
used to invade host
Hyaluronidase breaks tight junctions
Coagulase forms a clot around bateria for protection in blood stream
Exotoxins
1) A-B Toxin
B subunit binds to host cell receptor
A subunit mediates enzymatic activity responsible for toxicity
2) Membrane-Disrupting
Hydrolytic enzymes form pores in PM
3) Superantigens
bind to TCRs, regardless of specificity, and form bridge to MHC-II of APCs -> polyclonal activation
overwhelming release of Cytokines -> Cytokine Storm
Endotoxins
structural component of bacteria, NEVER SECRETED
Lipopolysaccharide (LPS) or Endotoxin
component of the outer membrane of Gram Negative bacteria
Lipid A component of LPS results in Endotoxin Shock if too much is released
Bacterial Genetics
Quorum Sensing = the ability to establish cell-cell communication
a critical mass or population density
too small pop -> signal molecules between Bacteria will just diffuse away before they can act
Genetic Diversity
1) Genetic Recombination
Homologous Recombination
gene REPLACEMENT via Transformation, Hfr Conjugation, Transduction
stable gene transformation
only homologous regions are incorporated, other regions are degraded
Site-Specific Recombination
requires only small region of homology, and restriction endonucleases (site-specific recombinases)
gene INSERTION via Transposition
ALL of the linear DNA is integrated - NO DNA is lost
2) Antigenic Variation
expression of alternate genes -> mutation or uptake of new info
Phase Variation = switch genes on/off (Phase on/off)
Cassette Switching (Site-Specific Inversion) = DNA is removed and reinserted in reverse direction to regulate expression
switching between 2 different Flagellin genes
Phase 1/2
3) Gene Transfer
1) Transposition
Transposons (jumping genes) insert via Site-Specific Recombination (non-homologous)
random movement, not capable of self replication, can be accompanied by duplication
2) Transformation
uptake of naked DNA by Gram+
cells must be COMPETENT (have Competence Factor)
must undergo Homologous Recombination after uptake, or else degraded
3) Conjugation
transfer of DNA via cell-cell contact, mediated by Sex Pili (F Pili), used by GramF Factor Plasmid
Tra Operon = Sex Pilus gene
oriT = where strand breakage occurs
Insertion Sequence = allows for Site-Specific Recombination
Donor cells = F+ (male)
Recipient cells = F- (female)
after Standard Conjugation, both cells are F+ (male)
Standard Conjugation
1) Donor cell encodes Sex Pilus
2) contact -> Conjugation tube links bacteria
3) F plasmid in donor is nicked at oriT
4) linear plasmid is transferred to recipient, then recircularized
5) both bacteria are now F+ Donors
NO bacterial chromosomal DNA is transferred, it is just the F plasmid
there is NO chromosomal alteration afterwards either
High Frequency Recombination (Hfr) Conjugation
1) insertion sequence on the F plasmid -> integration into Donor chromosomal DNA
once this happens -> bacteria is called High Frequency Recombinant (Hfr)
Hfr F+ Donor
2) strand breakage at oriT, which means bacterial chromosome becomes linearized too
3) b/c of its length, entire DNA strand (bacterial chromosome w/ integrated F plasmid) is not fully transferred
4) Recipient never recieves Tra Operon at the end of the strand
5) after transfer, some genes from Donor get incorporated into Recipient
6) however, b/c Recipient never received Tra Operon, it cannot make Sex Pilus, thus it remains F- Recipient
there IS chromosomal alteration of Recipients DNA
donor chromosomal genes that get integrated are those closest to the Integration Site on the F plasmid
4) Transduction
mediated by Bacteriophage (virus that infects bacteria specifically)
Generalized Transduction = Lytic Phage
transfer of bacterial genes from a disrupted cell = Lytic Cycle
Lytic Infection via Lytic Phage = Generalized Transduction
1) Bacteriophage injects viral DNA, then replicates in host
2) Bacterial chromosome gets digested
3) viral proteins assemble progeny
4) viral DNA is packaged into progeny
sometimes Bacterial DNA is packaged accidentally instead
5) lysis of cell -> release progeny
6) new phage with Bacterial DNA will inject a different Bacterium
7) Homologous Recombination is required for stable Transduction in this new Bacterium
Specialized Transduction = Temperate Phage
transfer of specific genes after integration of phage genetic material into bacterial chromosome = Lysogenic Cycle
Lysogenic Infection via Temperate Phage = Specialized Transduction
1) after infection, viral DNA is incorporated into host DNA, where it can lay dormant
2) when incorporated viral DNA is replicated to make phage progeny, adj bacterial DNA will go with it
only those genes adj to viral DNA will be Transduced
3) when that new phage injects a new host, it will inject bacterial DNA along with its phage DNA
4) this phage is defective, b/c missing some of viral genome
instead, it has a fragment of the original hosts DNA
5) new host integrates bacterial DNA via Homologous Recombination, and degrades viral DNA
Viruses
obligate intracellular parasites
Capsid + Viral Genome = Nucleocapsid
some have an Envelope = lipid bilayer coat acquired from host
Enveloped viruses do not kill their host
Naked Viruses = NO envelope, and KILL the cell they infect
Peplomers = viral glycoprotein spikes that act as receptors
Types of Viral Proteins
1) Structural = part of virion
2) Non-structural = NOT part of virion, used for replication of viral genome and production of Structural proteins
Viral Infections
Abortive
no viral progeny is produced
Nonpermissive and Nonproductive
Productive Non-Lytic
classically via Envelope virus
host cell is altered, but not killed
Permissive and Productive
Productive Lytic
classically through Naked virus
host cell death for release of progeny
Permissive and Productive
Latent
persistance with NO viral progeny
viral genome may integrate into host chromosome
can get reactivated from latency to Productive Lytic or Productive Nonlytic
Virus Life Cycle
1) Adsorption/Attachment = virus binds receptor on host cell
Tropism
M-Topic HIV binds Macrophages via CCR5
T-Tropic HIV binds Th cells via CXCR4
Heamagglutinin
2) Entry
Surface Fusion
NEUTRAL pH
formation of Syncytia (multinucleated cells)
Endosome Fusion
ACIDIC pH
virus fuses with membrane of endosome on the inside
Endosome Lysis
virus gets endocytosed and then lyses the endosome
Pore Formation
virus enters in vesicle, forms pores, genetic materal exits via the pores
Replication Sites
DNA viruses replicate in nucleus
RNA viruses replicate in cytoplasm
Exceptions:
Poxvirus = DNA virus that replicates in the cytoplasm
Poxvirus includes Smallpox (Variola), Vaccinia
Orthomyxoviruses = RNA virus that replicates in the nucleus
Orthomyxoviridae includes Influenza
Retroviruses = RNA viruses with DNA intermediate that replicate in BOTH the nucleus and cytoplasm
includes HIV, Hepatitis B
Types of Viruses and minimum required packaged components
Type I = dsDNA
Capsid + Viral Genome
no packaged or encoded enzymes
host cell provides everything needed
Type I Exception = Poxvirus
dsDNA, but replicates in Cytoplasm
Capsid + Viral Genome + DNA-dependent RNA Pol that works in cytoplasm
Encoded = DNA-dep DNA Pol that works in the cytoplasm is encoded for
Type II = ssDNA
Capsid + Viral Genome
no packaged or encoded enzymes
host cell provides everything needed
Type III = dsRNA
Capsid + Viral Genome + RNA-dep RNA Pol (to transcribe mRNA from the dsRNA)
Encoded = NONE
Type IV = (+)sense ssRNA
Capsid + Viral Genome
Encoded = RNA-dep RNA Pol to replicate genome
Type V = (-)sense ssRNA
Capsid + Viral Genome + RNA-dep RNA Pol (to transcribe -sense RNA to +sense RNA)
Encoded = packaged RNA-dep RNA Pol does it all
Type VI = (+)sense ss RNA, with DNA Intermediate (HIV)
Capsid + Viral Genome + Reverse Transcriptase (RNA-dep DNA Pol) + Integrase (incorporates viral DNA into host DNA)
Encoded = NONE (packaged enzymes do it all)
Type VII = partially dsDNA, with RNA Intermediate (HBV)
Capsid + Viral Genome + DNA-dep DNA Pol (to fill in the gap first)
Encoded = Reverse Transcriptase (RNA-dep DNA Pol), to make the partially dsDNA from the mRNA
Segmented Genomes
viruses with genomes made up of FRAGMENTS of RNA or DNR -> genotypic mixing
one host cell infected with 2 strains of segmented virus
new viral progeny could have a mixture of the 2 genomes = Genotypic Mixing
Phenotypic Mixing
genome does NOT change
if 2 different Enveloped viruses infect the same host cell
both types of Peplomers will be put on the surface
new viral progeny can have both types of peplomers when it buds off
Pseudotypes = progeny viruses with different Peplomers compared to the parental viruses
Plaque Assay
determines concentration of virus per mL
cells that get infected with virus will lyse and leave behind a plaque that does not stain
need to titer out virus, so that plaques are separate
at this point, 1 virion corresponds to each plaque
Amount of Virus/mL = (# plaques)(dilution)(1 / mL plated)
counted 23 plaques in a dilution of 10^5, and plated 0.1mL
(23)(10^5)(10) = 23 x 10^6 = 2.3 x 10^7
Diagnostic Inclusions
Rabies Virus -> Negri Bodies
eosinophilic inclusion bodies in CYTOPLASM of nerve cells
Cytomegalovirus (CMV) -> Owl Eye Inclusion Bodies
Basophilic inclusion bodies fill up the entire NUCLEUS
Hemagglutination Assay
used to determine Viral Titer = the lowest concentration of virus that still infects cells
Hemagglutinin cross links RBCs -> prevents them from collecting at the bottom of the well
tiny red dot in center of well = NO Hemagglutination
the last well where you see NO red dot = Viral Titer
Viral Titer = inverse of the Dilution
Hemagglutination Inhibition Assay
used to determin antiviral ANTIBODY Titer
look for the highest dilution concentration at which Hemagluttination is INHIBITED
the last well with a tiny red dot = the Titer of antibody
Hemadsorption = ability of cells infected with enveloped virus containing Hemagglutinin in its envelope to adsorb RBCs
Fungi
eukaryotic, unicellular/multicellular, heterotrophic (acuire nutrients from host)
most fungal infections are opportunistic (in immunosuppressed individuals)
modes of fungal growth
monomorphic (yeast)
unicellular, one form, reproduce asexually
multimorphic
multicellular, multiple forms
Hyphae = long chains which may be Septate or Aseptate
Septate = individual cells within Hypae are separated by walls
Parasites
Reservoir = hosts ensure continuity of parasites life cycle and act as sources of infection
Definitive Host = adult or sexual stages occur
Intermediate host = parasite lives larval and asexual stage
Vector = an organism that transmits parasite
Mosquito carries Malaria parasite
Protozoans
based on mobility
1) Amoebas
2) Flagellates
3) Apicomplexa = non-motile
4) Ciliates
Plasmodium spp Malaria
Obligate Parasite
Vertebrate Host = asexual phase (Merozoites)
Arthropod Host = sexual phase (Gametocytes)
Helminths
Flatworms = Tapeworms
Roundworms = Nematodes
Disease is consequence of the burden of the organisms in the host
Sensing Danger
PAMP = Pathogen-Associated Molecular Pattern
PRR = Pattern of Recognition Receptor
Antimicrobial proteins
Lysozyme degrades Peptidoglycan (effective against Gram+)
Defensins
alpha-defensins = produced by PMN (esp Neutrophils)
beta-defensins = produced by epithelial cells
Cathelicidins = produced by epithelial cells, macrophages, neutrophils
Major PRRs and their PAMPs
PRRs bind to PAMPs -> induce NFkB -> pro-inflammatory cytokine production
TLR1/2 = binds Lipopeptides and GPI
TLR2/6 = Lipoteichoic Acids and Zymosan (Gram+)
TLR2 = Chitin (fungal cell wall)
TLR4 = Lipopolysaccharide (outer plasma membrane of Gram-)
TLR5 = Flagella
Dectin-1 = Glucan
TLR9 = in endosome, binds Unmethylated CpG-rich DNA
TLR3 = in endosome, binds viral dsRNA
TLR7 = in endosome, binds viral ssRNA
TLR8 = in endosome, binds viral ssRNA
RIG-1 = binds cytoplasmic viral Nucleic Acid
NOD1, NOD2 = binds lipids of INtracellular Bacteria
PRR binds to PAMP -> induces production of Type-I Interferon (IFN-I)
IFN-alpha and IFN-beta
1) IFN-1 is produced by any NUCLEATED, infected cell
2) spreads to adj non-infected cells
3) induces expression of RNaseL, Protein Kinase R, and 2-5-Oligoadenylate Synthase
confers anti-viral state
RNaseL = mRNA degradation
Protein Kinase R = inhibition of protein synthesis
Oligoadenylate activates RNaseL
4) induces MHC-I expression
5) activates NK cells to release cytotoxic granules into infected cells
Cytokines
Properties
Pleiotropism = one cytokine has many different effects
Redundancy = 2 or more cytokines mediate similar functions
Synergy = combined effects of 2 cytokines is greater than additive individual effects
Antagonism = one cytokines inhibits another
Categories
Interleukins
signaling via Jak-STAT pathway
Chemokines
chemotactic activity
Interferons
antiviral activity
Complement (C)
Opsonization = tags an antigen for phagocytosis
C3b, C5b, etc
Encapsulated pathogens are resistance to phagocytosis, unless they are opsonized
Classical Pathway
1) Ab binds to Ag
2) C1q binds Ab bound to Ag
3) activation of C1s -> generation of C3 Convertase and C2b
C2b causes vessels to become leaky
b complements usually bind, C2b is the exception
4) C3 Convertase cleaves C3 -> C3a and C3b
5) C3b opsonizes, and C3a attracts phagocytes
6) C5 Convertase claves C5 -> C5a and C5b
7) C5b starts formation of MAC complex (C5,6,7,8,9), and C5a is the most powerful chemoattractant
Adaptive Initiation of Classical Pathway
C1 binds to C-Reactive Protein (CRP) on pathogen surface -> then get activation of regular pathway
instead of binding Ab
Lectin Pathway
1) Mannose-Binding Lectin (SOLUBLE PRR) binds to Mannan on pathogen surface
2) activation of MASP-2 -> generation of C3 Convertase, and C2b
C2b causes vessels to become leaky
3) C3 Convertase cleaves C3 -> C3a and C3b
4) C3b opsonizes, and C3a attracts phagocytes
5) C5 Convertase claves C5 -> C5a and C5b
8) C5b starts formation of MAC complex (C5,6,7,8,9), and C5a is the most powerful chemoattractant
C3 Convertase of the Classical and Lectin Pathways
C4b2a
C5 Converase of the Classical and Lectin Pathways
C4b2a3b
Alternative Pathway
1) Spontaneous tick over of C3 to generate C3b
2) C3b binds pathogen surface
3) Factor B binds and then is cleaved by Factor D -> Bb and Ba
4) Bb binds to C3b -> C3bBb = C3 Convertase of the Alternative Pathway
5) lots of C3b is made, so some will bind to C3 Convertase -> C3bBb3b = C5 Convertase
5) C5b opsonizes, and C5a is the most powerful chemoattractant
MAC = Membrane Attack Complex
inserts across plasma membranes
Susceptible pathogens include:
Gram Negative bacteria, Enveloped viruses
made up of C5b (initiator) + C6,7,8, and multiple C9 units
Resistant pathogens :
Gram Positive (Lysozyme deals with these)
Encapsulated pathogens (opsonization deals with these)
Complement Receptors
CR1, CR3, and CR4
receptors important for opsonization and stimulate phagocytosis of pathogen
CR1 is important for cleance of Immune Complexes
Regulatory C Proteins
C1-INH = C1 Inhibitor
inhibits Lectin and Classical Pathways
Hereditary Angioedema = C1-INH deficiency -> excessive levels of C2b, which causes vessels to become leaky -> edema
DAF, MCP, C4BP, and CR1
all disrupt C3 Convertase formation
CD55 (DAF) and CD59 block MAC assembly on self-cells
deficiency of CD55 and/or CD59 -> lysis of RBCs -> Nocturnal Hemolysis
Split Products Generated
C3b = opsonization
C5a > C3a >>> C4a = leukocyte chemoattractants
C5a > C3a >> C4a = increase vascular permeability
C5a and C3a = Mast cell activation and degranulation
activated Mast cells release pro-inflammatory mediators
Histamine = vasoDILATION and vascular permeability (SHORT-LIVED)
TNF-alpha and IL-1 = vascular permeability (SUSTAINED)
IL-8 = chemoattractant
LTB4 = chemoattractant
Prostaglandins = vasoDILATION
fluid that seeps out of leaky vessels contains Bradykinin
Bradykinin stimulates PAIN and Mast cell degranulation
PRRs binds to PAMPs cause Macrophages to release Cytokines
IL-1 activates vascular endothelium by making them sticky (adhesion molecules)
IL-6 induces Acute Phase Protein production
C proteins, MBL, CRP
Innate Response and Inflammation
Cardinal Signs of Inflammation
Rubor = Redness
Tumor = Swelling
Calor = Heat
Dolor = Pain
Functio Laesa = Loss of Function
Cell-adhesion Molecules
Selectins
E-selectins on Endothelium bind to Sialyl LewisX (CD15) on Neutrophils -> rolling
Integrins
LFA-1 (CD18) on neutrophil binds ICAM-1 of activated endothelium -> diapedesis
Extravasation
1) Rolling
SialylLewisX (CD15) on Leukocyte binds E-Selectin on endothelium
2) Tight Binding
LFA-1 (CD18) on leukocyte binds ICAM-1 on endothelium
3) Diapedesis
squeezing in between endothelial cells
4) Migration
towards chemokine gradient
Neutrophil Killing
kill via Phagocytosis
1) Oxidative Killing
NADPH Oxidase -> Superoxide Dismutase -> Myeloperoxidase
process makes HOCl (Hypochlorite)
gets released into bacteria
Oxidative (Respiratory) Burst = Superoxide (O2-) is produced via NADPH
consumes a LOT of Oxygen
Inflammation
Acute = dominated by Neutrophils
Chronic = dominated by macrophages (and some Th1 cells activating them)
Granuloma = center of macrophages with endocytosed pathogen, surrounded by wall of Th1 cells
Mycobacterium Tuberculosis
Granulomas form after several weeks
Killing By NK cells
1) Perforins/Granzymes
2) FasL on NK cell with Fas on target cell

B2 Review
Saturday, December 13, 2014 at 2:36 PM

Inducers and Targets

Epitope (antigenic determinant) = exact region of contant between Ag and receptor
TCRs only recognize short LINEAR epitopes of polypeptides, presented on MHC molecules by APCs
Dentritic cells are naturally APCs
activated B cells and Macrophages become APCs
B cell Epitopes vs T cell Epitopes
Receptor?
MHC required?
binds SOLUBLE Ag?
characteristics of Ag?
epitope properties

B cells
Membrane Ig (BCR) -> Ag
NO
YES
proteins, polysaccharides, lipids,
nucleic acids
linear or conformational

T cells
TCR -> Ag/MHC
YES
NO
proteins mostly, some glycolipids
short linear peptides ONLY

B-cells require cross-linked of their BCRs to become activated

therefore, soluble epitopes released by the Ag will NOT cause B cell activation
monovalent Ags will NOT cross-link B cells
multivalent Ags = many different epitopes, or an identically repeated epitope
provides numerous epitopes for BCRs to bind and cross-link (group) together
Hapten = small Ag that is NOT immunogenic on its own
Hapten-Carrier Conjugate is immunogenic
Penicillin
Penicillin/self-protein adducts elicit an immune response (Hypersensitivity reaction)
Immunoglobulin (Ig) Structure
2 chains:
Heavy Chain
Light Chain
2 binding sites:
Variable
Constant
2 forms:
Membrane-bound (BCR)
Secreted
each B cell has a unique BCR is expresses
eventually secretes its BCR as Abs
there are 2 binding sites for Abs and therefore BCRs

Fab = Ag Binding Fragment

region is highly variable from Ab to Ab, so NO crystal is formed in vitro
Fc = Crystallizable Fragment (Constant)
all Abs in the solution have very similar AA sequence, thus forms crystal in vitro
Hinge Region = increases Avidity

each variable domain of a chain has 3 Hypervariable regions

Complementarity Determining Regions (CDRs)
CDR3 is the most hypervariable (contains N and P nucleotide addition)
a total of 6 CDRs make up the Ag binding site on ONE arm
therefore each Ab has 12 CDRs
the tighter the CDR binds = the higher the Affinity
Avidity = binding due to a sum of the affinities of several sites
2 binding sites = sum will be greater than binding of each site individually
IgM has the highest Avidity
less IgM is required for Cq1 binding and Classical C activation
IgM is the best activator of Complement
Ig Classes
Membrane Ig
integrated in the Plasma Membrane of the cell that synthesized it
this is the BCR
Circulating Ig
circulating in the blood
Secretory Ig
from tissues where it was made, to the lumenal side of mucosal epithelium (GI, respiratory, etc)
Cell-bound Ig
cell has Fc Receptors and will grab the Ab
Mast cells have Fc-e-R which binds to IgE -> armed Mast cell
Classes of Immunoglobulins
IgG
Circulating Monomer
most abundant
best for Opsonization (Fc region is easily recognized by phagocytes)
Transplacental
Brambell Receptor or Neonatal Fc Receptor (FcRn)
IgM
Membrane and Circulating Pentamer
the BCR, and first class of Ab released by B cells
the best activatory of the Classical Pathway
highest Avidity
IgD
Membrane Monomer
co-expressed with IgM due to differential RNA splicing
IgA
Circulating Monomer, OR Secreted Dimer
Mucosal protection, and in colostrum and breast milk
Poly-Ig Receptor (pIgR)
binds to and carries IgA across mucosal epithelial cells to get to lumenal side
binds specifically to J Chain (therefore IgA and IgM)
IgE
Circulating or Cell-bound Monomer
picked up by Mast cells and Basophils via high affinity Fc-e-R -> arming
parasitic infections and allergies
the only class of Ig that has an Fc region that binds before it binds to Ag
identify what type of Ig patient expresses
everyone has been exposed to Ragweed Pollen
therefore all patients express IgG against it
however, those who are allergic to Ragweed Pollen ALSO expres IgE against it
if you detect IgG to Dengue in a patient, all that tells us is that pt has been infected with Dengue in their lifetime
if you detect IgM or IgM + IgG to Dengue, then you know it is a CURRENT infection

Isotypic Abs = Anti-Class Abs

Class-specific
Abs bind to the Fc region of foreign Abs
Allotypic Abs = recognize individual variation
each individual has slightly different Fc regions
Idiotypic Abs = Idiosyncratic Abs
Abs bind to the Variable Region of foreign Abs

Ig Genetics
B cells
1) CD34+ Common Lymphoid Progenitor
2) IL-7R on CLP with IL-7 on Bone Marrow Stromal cell
3) c-Kit on CLP with SCF (Stem Cell Factor) on Bone Marrow Stromal cells -> commits to B cell lineage
4) Heavy Chain rearrangement
VDJ Somatic Rearrangement
RAG1/2 and RSS Sequences (12/23 Rule = flanking 12 must combine with 23)
5) functional rearrangement induces Allelic Exclusion
other H chain allele gets turned off (assuming the first one was successful)
get 2 tries to make a good H chain
H allele 1
H allele 2
6) Pre-B cell with Pre-BCR
good Heavy Chain gets put with Surrogate Light Chain
7) activating signal via ITAM (Ig-beta and Ig-alpha)
8) Proliferation (Clonal Expansion)
it is hard to make a functional heavy chain
thus after Pre-BCR -> B cell undergoes clonal expansion
each of those daughter cells then undergoes its own L chain rearrangement
increases the odds that one of them will be able to make a functional light chain
9) Light Chain Rearrangement
VJ Somatic Rearrangement
RAG1/2, RSS Sequences (12/23 Rule)
10) functional rearrangement induces Allelic Exclusion
other L chain alleles get turned off
if first try was successful, will turn off the other kappa allele, and the two lambda alleles
have a total of 4 trys to make a good light chain
kappa allele 1
kappa allele 2
lambda allele 1
lambda allele 2
11) Immature B cell
expresses only IgM
12) Tolerance = test Immature B cells for auto-reactivity
if B cell binds to self-antigen:
1) Receptor Editing
RAG1/2 is re-expressed, and Light Chain gene undergoes Somatic Rearrangement again
if new BCR is no longer auto-reactive, cell is allowed to leave BM
2) Apoptosis
if after Receptor Editing, B cell is still auto-reactive -> Apoptosis
3) Anergy = INactivation/paralysis
if BCR binds to SOLUBLE self-antigen
13) B cells leaves Bone Marrow
14) Alternative RNA Splicing of Tandem Constant region genes
will get IgD co-expressed with IgM
RNA Splicing is what combines Constant Region with Variable Region
Constant Region is already encoded for in genome, thus does NOT undergo DNA rearrangement
Primary RNA transcript includes the rearranged Variable Domain, Introns, and the Constant Domain
Splicing creates an mRNA with these sequences adjacent
Somatic Rearrangement
DNA event
1) Recombinatory Signal Sequences (RSS) are brought together by RAG1/2
DJ is first
12 RSS flanking D is brought to 23 RSS flanking J
RAG1/2 cleaves RSSs off, then generates DNA hairpins at the coding ends
2) Artemis opens DNA hairpins -> generates Palindromic P-Nucleotides
3) N-nucleotides are then added on by TdT
Terminal Deoxynucleotidyl Transferase
4) strands are paired
unpaired nucleotides are removed by exonuclease
5) gaps are filled in by Ligase
just the joining of D to J and V to DJ alone is called Combinatorial Diversity
there are many different Vs, Ds and Js to choose from
the addition of P- and N-nucleotides = Junctional Diversity
CDR3 encompasses the joining regions between V, D, and J
therefore CDR3 is the most Hypervariable of the Hypervariable Regions, b/c it includes the Junctional Diversity
CDR1 and CDR2 are fully encoded within the V gene segment
P- and N-nucleotide additions add lots of diversity
however, high chances of adding a frameshift, premature stop codon, etc
this is why most B cells never make it to the Pre-BCR stage, let alone Immature B cell stage
also why it gets so many tries to make a functional heavy and light chain
X-linked Agammaglobulinemia
Brutons Tyrosine Kinase (btk) deficiency
NO activating signal from Pre-BCR
ALL B cells arrest at Pre-B cell stage
therefore NO circulating CD19+ cells (B cells)
B cell Activation
1) BCR on Mature B cell binds to Ag
2) cross-linking of BCRs
Capping = merging of BCRs onto lipid rafts, and those rafts coalescing together
3) Activation Pathway
1) activation of Src Kinases (Lyn)
CD45 Phosphatase removes inhibitory phosphate from Src Kinases
2) phosphorylation of ITAMs
3) recruit and activate Syk
4) activate PLC-gamma
5) PIP2 -> DAG + IP3
6) activation of MAPK, and increase in intracellular Ca2+
7) activation of PKC and Calcineurin
8) activation of TFs
PKC -> NF-kB
MAPK -> AP-1
Calcineurin -> NFAT
4) Clonal Expansion
B cell Positive Co-receptor = CD21, CD19, CD81
decreases threshold for immunogen activation of B cell
1) CD21 binds to C3b on pathogen surface
2) CD19 becomes phosphorylated
3) phosphorylated CD19 binds Src family Kinases and PI3K
4) initiation of downstream signaling
B cell Negative Co-receptor = Fc-gamma-R
overrides Positive Co-receptors
1) Fc-gamma-R binds to Ab that is already bound to Ag
2) ITIM (Inhibition Motif) becomes activated
3) Inhibitory signal starts negative signal cascade
dont need a cop to arrest someone already in handcuffs
Thymus-INdependent Antigens
NO protein domains, but has repeated epitopes
Polysaccharides, Nucleic Acids, etc
T cells can NOT recognize these Ags, thus no T cell help
Steps a B cell undergoes with NO T cell help
1) BCR binds Ag
2) Clonal Expansion
3) production of IgM
NO isotype switching though, so ONLY IgM
4) production of co-stimulatory molecules (CD80/86)
to activate T cells
5) production of Cytokine Receptors
to receive cytokine help from Th cells
only activated B cells therefore are able to receive help from Th cells
resting B cells do not express these receptors, so they just sit idle
6) Migration
B cells move toward T cell zone (PALS or Paracortex)
Characteristics of Thymus INdependent Antigens
TI-1 Antigens
Lipopolysaccharide is major example
Mitogen = binds to a cell and causes clonal expansion (activate ALL cells, regardless of specificity)
LPS binds to TLR4 on ALL B cells -> activates ALL B
Nonspecific, Polyclonal response
TI-2 Antigen
mutlivalent or repetitive epitopes
repeated epitopes cause cross-linking of BCRs -> activation
C3b on pathogen surface binds to Positive Co-receptor and helps
B cell Types
B-2 cells
the conventional B cells
made in BM and main location is secondary lymphoid tissues (lymph nodes, spleen)
B-1 cells
have CD5 marker (which B-2 cells do NOT)
self-renewing, and main location is in body cavities (peritoneal, pleural)
able to make Ab with NO T cell help (since Th cells cant get to them)
B cells activated by Thymus Dependent Antigen
Signal 1 = BCR binds to Ag
Positive Co-receptors also bind to C3b opsonin
B cell becomes APC
expresses co-stimulatory molecules (CD80/CD86), which bind to CD28 on T cell
Signal 2 = CD40L on T cell binds to CD40 on B cell
1) Germinal Centers
clonal expansion -> accumulation of activated B and T cells together in one place
2) Somatic Hypermutation
Activation-Induced cytidine Deaminase (AID) deaminates DNA at Cytosine residues
repair of deaminations results in permanent base changes
3) Affinity Maturation
clonal selection of those BCRs with highest affinity for Ag, after SHM changes
Signal 3 = CD40L on T cell binds CD40 on B cell (again)
1) Isotype Switch
binding of cytokine determines class
IL-4 binding -> IgE
IL-2 binding -> IgG
2) differentiation to Memory or Plasma cell
X-linked Hyper IgM Syndrome
CD40L deficiency
NO Germinal Centers
normal numbers of T and B cells
MHC Molecules
MHC class I
HLA-A, -B, -C
ONE polypeptide alpha chain + beta-2 Microglobulin (non-MHC protein)
expressed on ALL nucleated cells
CD8 binds to alpha-3 domain of MHC-I
alpha-3 domain is conserved, therefore NOT polymorphic
MHC class II
HLA-DP, -DQ, -DR
TWO polypeptide chains = alpha and beta chains
expressed on professional APCs
prevalent in the Thymus, as CD4+ T cell recognize Ag/MHC-II
Homozygous = will express 3 MHC-I
one type of HLA-A, one HLA-B and one HLA-C
3 MHC-II molecules on the surface of their cells
Heterozygous = will express 4-6 MHC-I
up to 2 different HLA-A, 2 HLA-B, 2 HLA-C
4-6 different MHC-II molecules on the surface of their cells
Hybrid MHC-II
since 2 chains combine to form MHC-II, there can be additional polymorphism
Cis = maternal alpha chain + maternal beta chain
Trans = maternal alpha chain + paternal beta chain
Peptide Binding Groove
MHC-I
CLOSED ends
8-10 AAs, strict limit
participating in binding = alpha 1 and alpha 2
Anchor Residues = ends of the groove/peptide
position of the ends of the peptide fragment = buried in the floor of the cleft
MHC-II
OPEN ends
13+ AAs
participating in binding = alpha 1 and beta 1
Anchor Residues = entire length of peptide fragment
position of the ends of the peptide fragment = protruding from the ends of the cleft
Anchor Amino Acid Residues
those AAs on the peptide that specifically bind to the MHC molecule in the groove
Anchor Residues are NOT involved in binding to the TCR
the residues of the peptide fragment that bind to the TCR, are the ones protruding up from the cleft
Antigen Processing and Presentation
MHC class I
ENDOGENOUS pathway
processes proteins made inside the cell
1) defective, aberrant, and/or viral proteins get degraded into peptides by Immunoproteasome
IFN-gamma induces expression of the Immunoproteasome
3 new subunits replace subunits on the Proteasome
IFN-gamma is produced by Th1 cells
Immunoproteasome specifically degrades proteins into peptides suited to bind to MHC-I
2) TAP brings these peptides into the rER
3) ERAP removes N-term AAs to give a peptide that is 8-10 residues
4) MHC-I alpha chain binds Calnexin,Calreticulin until beta-2 Microglobulin binds
5) alpha:microglobulin complex binds to TAP, via Tapasin
6) MHC-I molecule binds peptide of appropriate length and sequence, which was delivered by TAP earlier
7) this completes its folding, and MHC-I is released from TAP and exported to the cell membrane
MHC class II
EXOGENOUS pathway
processes proteins from outside the cell (Ags obtained by endocytosis)
1) endocytosis of extracellular protein
2) that endocytic vesicle fuses with an endosome
3) MHC-II is synthesized in the rER
4) Invariant Chain binds MHC-II, and blocks the binding cleft
Invariant chain also provides the trafficking signal to direct the MHC-II to the lysosome
5) MHC-II with Invariant chain is sent to an acidic lysosome
6) Invariant chain is immediately degraded to CLIP, which remains bound in cleft
7) lysosome fuses with the endosome holding the endocytosed antigenic protein
8) foreign protein is degraded by lysosome acidic proteases
9) HLA-DM removes CLIP from MHC-II
10) HLA-DM grabs different peptides and puts them in the now free binding cleft, until one fits
11) Lysosome fuses with plasma membrane, putting Ag/MHC-II on the surface
EXCEPTION = Dendritic cells
ONLY Dendritic cells are capable of Cross-Presentation
DCs can put Exogenous protein onto MHC-I
CD8+ T cells bind to exogenous Ag/MHC-I -> CTL kills that cell
also, DCs can put Endogenous protein onto MHC-II
another way of getting self-peptides onto MHC molecules
Viruses can block endogenous pathway
1) inhibit TAP, thereby inhibiting peptide transport into ER
2) inhibit peptide binding onto MHC
3) cause MHC-I degradation
Intracellular bacteria block exogenous pathway (Listeria)
1) escape endosomes
2) neutralize endosome acidification
3) block fusion with lysosome
4) sequesters MHC-II after vesicle fusion
CD1 = MHC-I like protein
presents LIPID antigens to NKT cells
NKT cells are T cells with a TCR, but they are specific ONLY to CD1

B3 Review
Saturday, December 13, 2014 at 8:44 PM

TCR and T cell

TCRs recognize LINEAR epitopes of PEPTIDES that are held by MHC molecules
2 main types of TCRs
alpha/beta TCR
alpha = light chain-like (VJ rearrangement)
beta = heavy chain-like (VDJ rearrangement)
gamma/delta TCR
Life of a T cell
1) Common Lymphoid Progenitor (CD34+ and CD44+) migrates from BM to Thymus
2) Notch Signal from Thymic Epithelial cell instructs CLP to commit to T cell lineage
3) IL-7R on CLP binds IL-7 on Thymic cell
4) c-Kit on CLP binds SCF on Thymic cell
5) beta-chain somatic rearrangement, and CD3 expression
RAG1/2, RSS (12/23 rule)
CD3 = Signal Transduction Complex, the equivalent of Ig-alpha and Ig-beta of BCR
6) co-expression of CD4 and CD8 = DP Thymocyte
7) Pre-TCR (beta chain + surrogate alpha chain + CD3)
8) activating signal by Pre-TCR
9) Allelic Exclusion
beta-chain allele 2, and gamma/delta genes are turned OFF
10) Proliferation (Clonal Expansion)
all daughter cells will undergo following steps
11) alpha-chain Somatic Rearrangement
delta chain gene is entirely within V gene segment of alpha chain, and will be excised by RAG1/2 in this step
12) Positive Selection
Cortex of Thymus, mediated by Cortical Thymic Epithelial cells
cTECs present Thymus-Specific self-antigen (they do NOT express Aire)
if TCR binds MHC-I -> CD8+ SP Thymocyte
if TCR binds MHC-II -> CD4+ SP Thymocyte
if TCR binds CD1 -> NKT cell
95% of T cells die by neglect = are not able to bind to MHC
13) Negative Selection
Central Tolerance
clonal deletion of auto-reactive TCRs
Medulla of Thymus, mediated by Medullary Thymic Epithelial cells
Aire is expressed by mTECs -> expression of ectopic Peripheral Tissue-Specific self-peptides
enables killing of those T cells that would react in periphery
if TCR binds with HIGH affinity to self-peptide/MHC -> apoptosis
if TCR binds with LOW affinity to self-peptide/MHC -> allowed to leave as a SP T cell
if TCR binds with MODERATE affinity to self-peptide/MHC -> Natural T Regulatory Cell
Treg = CD4+, CD25+, Foxp3+
14) Peripheral Tolerance
mature, self-reactive T cells that escaped Central Tolerance are killed, suppressed, or made anergic
Treg cells express TGF-beta and IL-10
down-reg of CD80/86 on APCs -> anergy of T cell
also inhibits T cell proliferation
Structure of the TCR
Heterodimeric
95% of T cells have alpha/beta TCR, while 5% have gamma/delta TCR
N terminus Variable Domain is responsible for contact with the Ag
CDR3 is the most hypervariable (generated from P and N nucleotide addition - Junctional Diversity)
CDR3 interacts specifically with the Ag peptide in the cleft (the parts of the peptide sticking out)
CDR1 and CDR2 are both encoded entirely within the V gene segment, and specifically make contact with the MHC molecules arms
bind to alpha-1 and beta-1 of MHC-II, and alpha-1 and alpha-2 of MHC-I
CD4 and CD8
CD8 binds alpha-3 invariant (constant) region of MHC-I
CD8 is heterodimer = alpha/beta or alpha/alpha
CD4 binds beta-2 invariant (constant) region of MHC-II
CD4 is Monomeric
T cells vs B cells
T cells do NOT undergo Somatic Hypermutation
T cell maturation takes place in the Thymus, not the Bone Marrow
T cells undergo Positive and Negative Selection
gamma/delta T cell
appears in fetal Thymus
does NOT undergo Selection
therefore leaves Thymus as a Double Positive gamma/delta T cell
immunoserveilance
limited diversity
APECED/APS-1
Aire deficiency
autoimmune attack on multiple endocrine tissues
due to a failure of Negative Selection by mTECs
Naive T cell Activation
1) LFA-1 on T cell binds ICAM-1 and ICAM-2 on APC with LOW affinity
2) CD2 on T cell binds CD58 (LFA-3) on APC, for adhesion
3) Clonal Selection
I) Signal 1 = TCR-CD3 binds to Ag/MHC
Signal 1 in T cell activation
II) LFA-1 undergoes conformational change to high affinity for ICAM on APC
tighter adhesion to APC
III) Activating Signal Pathway
1. CD45 Phosphatase removes inhibitory phosphate from Lck (Src Kinase)
2. Lck phosphorylates CD3 ITAMs
3. CD3 ITAMs recruit ZAP-70
4. Lck phosphorylates ZAP-70
5. ZAP-70 recruits PLC-gamma
6. PLC-gamma activates after Signal 2 -> cleaves PIP2 -> IP3 and DAG
IP3 increases ICF Ca
Ca and DAG activate PKC and Ras, MAPK
7. Ca activates NFAT
8. PKC activates NF-kB
9. Ras, MAPK activate Fos, AP-1
10. these TFs induce IL-2 production and IL-2R expression
IV) Signal 2 = CD28 on T cell binds CD80/86 on APC
IL-2 production, and IL-2R expression
CD25 = alpha subunit of high affinity IL-2R
low affinity IL-2R does NOT have alpha subunit
4) Clonal Expansion
IL-2 binds IL-2R via autocrine and paracrine mechanism
5) Signal 3 = binding of specific cytokine -> differentiation into Th subgroup
Outcomes of T cell Activation Signals
Signal 1+2 -> activation of T cell
production of IL-2 and IL-2R
Signal 1 alone -> T cell becomes anergic (INactivated)
Signal 2 alone -> no effect (b/c there is no Ag component)
Effector T cells ONLY REQUIRE SIGNAL 1
naive T cells that require Signal 1 + Signal 2
Inducers of T cell activation
Foreign Ag
activation of 1/100,000 (0.001%) of T cells
Allogenic MHC = foreign MHC, typically with transplants
activation of 1/100 (1%) of T cells
1) TCR binds only to the foreign MHC molecule
CDR1 and CDR2
2) TCR binds only to the antigenic peptide held by the foreign MHC molecule
CDR3 (the hypervariable domain with the greatest specificity
Superantigens
activation of up to 1/5 (20%) of T cells
b/c it is INdependent of binding specificity of the TCR
bacterial exotoxins
bind and cross-link MHC-II
Staph Enterotoxin binds to alpha/beta TCRs, regardless of their specificity, and form a bridge between TCR and MHC
overstimulation of T cells systemically -> cytokines torm
Toxic Shock Syndrome Toxin
Polyclonal, in vitro stimulation = non-physiological stimuli
PHA, ConA = Plant Lectin Mitogens
Phytohemagluttinin (PHA) binds CD2 -> widespread polyclonal activation of T cells
Anti-CD3 and Anti-CD28 mAbs
Signal 1 and Signal 2 -> activation of T cells
PMA and Ionomycin
PMA mimics DAG -> activates PKC
Ionomycin forms pores for Ca2+ influx
Long-Lived Memory T cells doe NOT require Signal 2 anymore
they only require Signal 1 = TCR-CD3 binding to Ag/MHC
Contraction Phase
mediated by FasL/Fas
apoptosis of activated T cells, at the end of clearance of the pathogen
Effector T cell Types
CD8 = Cytotoxic T cells (CTL)
kills virus-infected cells
Cytotoxic Granules are released specifically at the point of contact between CTL and virus-infected cell
prevents excessive collateral damage
CD8+ cells become armed with Perforin/Granzymes after receiving IL-2 while CD8+ cell is bound to Ag/MHC-I
before then, naive CTL precursors do NOT have granules yet - incapable of killing
(we dont want them to kill the dendritic cell that activates them in the first place)
Killing Mechanisms
Perforins/Granzymes
FasL on CTL binds Fas on target cell -> Caspase cascade -> apoptosis
much slower b/c must wait for FasL expression
CD4 Th1 cells
INTRACELLULAR pathogens
activate macrophages, and attract more macrophages to the site of infection
deal with Intracellular Microbes
IL-12 and IFN-gamma -> T-bet
IL-12 is ONLY produced by APCs, and is the MAIN differentiation factor
Th1 produces IL-2 and IFN-gamma
IL-2 and IFN-gamma -> activated CD8 T cells to become Cytotoxic (CTLs)
IFN-gamma and expression of CD40L activates Macrophages to destroy engulfed bacteria
CD40L binds to CD40 on Macrophages -> activation of Macrophages
CD40L/CD40 also for B cell activation (2nd and 3rd Signal)
IFN-gamma -> Opsonization and activated B cells to isotype switch to IgG subclass
IL-2 -> major cytokine for the Proliferation of Activated T cells -> increasing numbers of effector T cells
CD4 Th2 cells
EXTRACELLULAR pathogens
promote antibody production
IL-4 -> GATA3
Th2 produces IL-4, IL-5, IL-10, and IL-13
IL-4 -> B cell class switch to IgE -> IgE production
primary antibody to combat parasitic worms (Helminths)
also SUPPRESSION of macrophage activation (opposite IFN-gamma)
IL-5 -> Eosinophil activation (effector cells that deal with parasitic worms)
IL-13 -> Mucosal secretions
IL-10 -> inhibit macrophage activation
CD4 Th17 cells
enhance neutrophil response (pro-inflammatory response), barrier functions, especially at mucus membranes
TGF-beta + IL-6 -> ROR-gamma-T
Th17 produces IL-17 ONLY
IL-17 is pro-inflammatory - acts on cells and tissues to produce a wide variety of pro-inflammatory cytokines
IL-17 also causes cytokine production that promotes Neutrophil production by the bone marrow
IL-8 is the most potent chemoattractant that we have (which is produced in response to IL-17 release)
Regulatory T cells (Treg)
maintain Peripheral Tolerance
suppression of a T cell by a Treg cell requires them to interact with the same APC
when both bound to APC -> Treg cell inhibits expression of CD86/80 by the APC -> anergy of T cell
TGF-beta (by itself!) -> Foxp3
recall that TGF-beta + IL-6 will cause the Th0 cell to differentiate into a Th17 cell
they are CD4+ CD25+ Foxp3+ Tregs
Treg produces TGF-beta and IL-10
IL-10 -> inhibits APCs and renders them unable to activate naive T cells, by preventing Signal 2
IL-10 blocks expression of CD80/86
IL-10 and TGF-beta -> inhibit macrophage activation
TGF-beta -> inhibits T cell proliferation
Antibody Dependent Cell-mediated Cytotoxicity (ADCC)
overlap of Humoral and Cell-mediated Immunity
1) Ab binds surface Ag on cell
2) NK cells, w/ their Fc Receptors, bind to Fc region of Ab bound to surface Ag
3) release of Perforin/Granzymes
Eosinophils do the same thing with IgE bound to parasitic worms
Cytotoxic Killing Mechanisms
Perforin/Granzymes
1) CTL binds Ag/MHC-I
2) release of Perforin/Granzymes into Immunologic Synpase
3) Perforin (resembles C9) makes pores
4) Granzymes enter through pores and induces production of Caspase-3
5) Caspase cascade -> apoptosis of infected cell
FasL/Fas (CD95L/CD95)
1) CTL binds Ag/MHC-I
2) FasL expression on T cell is induced following TCR binding
3) FasL binds Fas on target infected cell
4) Caspase-8 production in infected cell
5) Caspase cascade -> apoptosis
slower pathway, b/c must wait for FasL expression AFTER TCR binds Ag/MHC-I
Vaccines
Haemophilus Influenzae type b (Hib) vaccines
CONJUGATE Polysaccharide vaccine
has to be given as a conjugate, b/c Polysaccharide capsule is the antigen used, which would NOT elicit a Memory response on its own
Pneumococcal Vaccines
CONJUGATE Polysaccharide vaccine
pure Polysaccharide
Influenza (Orthomyxoviridae) Vaccines
FluMist (intranasal Flu Vaccine) = Live Attenuated
inactivated Influenza virus injection
Capsule Polysaccharide vaccine = Conjugate
Polio Vaccines
Oral Polio Vaccine (Sabin) = Live Attenuated
Salk Polio Vaccine = inactivated
Smallpox (Variola) vaccine
Cowpox/Vaccinia Virus = Live attenuated, confers protection against the human Smallpox
Measles, Mumps, Rubella (MMR)
MIXTURE of all 3 Live Attenuated viruses
Rabies Vaccine
Inactivated vaccine + passive immunization with Ab
DaPT vaccine
Mixture of Diptheria, Pertussis, and Tetanus Protein SUBUNIT Vaccine
Hepatitis B (Retroviridae) vaccine
produced by recombinant DNA technology = Subunit vaccine

List of Molecules
Monday, December 15, 2014 at 9:48 AM

List of CD Molecules
CD1 = MHC-I like molecule that holds lipid antigen and presents it to NKT cells (CD1-restricted T cell)
CD2 = on Lymphocyte, binds to CD58 (LFA-3) on APC
also, Phytohemagglutinin (PHA) binds to CD2 -> polyclonal, systemic activation of T cells
CD3 = Signal Transduction Complex of the TCR
CD4 = Helper T cells, binds to beta-2 invariant region of MHC-II
CD5 = B1 cell marker
CD8 = Cytotoxic T cells, binds to alpha-3 invariant region of MHC-I
CD14 = binds Lipopolysaccharide (Endotoxin), just like TLR-4
CD15 = (Sialyl LewisX), binds to E-Selectins on Endothelium -> Rolling
CD18 = (LFA-1), binds to ICAM-1 on Endothelium -> tight adhesion -> diapedesis
CD19 = pan B2 cell (typical B cell) marker, part of the Positive Co-receptor
gets phosphorylated after CD21 binds C3b on the pathogen surface
CD20 = pan B cell marker
anti-CD20 (Rituxan) is used to treat B cell Lymphoma
CD21 = B cell Positive Co-receptor component that specifically binds to C3b on the surface of the pathogen
CD25 = the alpha chain of the high affinity IL-2 Receptor (IL-2R)
CD28 = on T cell, binds to co-stimulatory molecules CD80/86 (B7) on APC -> 2nd T cell activation Signal
CTLA-4 = on T cell, outcompetes with CD28 for binding to CD80/86, and is inhibitory -> inactivation of T cell
CD34 = Hematopoietic Stem Cell (HSC) marker
CD40 = on B cell, binds to CD40L on Helper T cell -> 2nd and 3rd B cell activating signal
CD40L = on Helper T cell, binds to CD40 on B cell -> 2nd and 3rd B cell activating signal
also binds to CD40 on Macrophage -> increased killing
CD44 = Common Lymphoid Progenitor (CLP) marker
CD45 = Phosphatase activity -> removes inhibitory phosphate from Src Family Kinases (eg Lck)
CD55 = DAF, inhibits MAC lysis of self-cells
CD56 = pan NK cell marker
CD58 = (LFA-3) on APC, binds to CD2 on T cell
CD59 = inhibits MAC lysis of self-cells (along with DAF/CD55)
CD81 = part of the B cell Positive Co-receptor, the Transmembrane spanning domain
CD95 = Fas on cell to be killed, binds to CD95L on CTL or NK cell -> Procaspase 8
CD95L = FasL on CTL or NK cell, binds to CD95 on cell to be killed -> Procaspase 8