B-cells

Follicular B2 cells
-​ Require T cells for activation (need memory cells)
-​ TDependent antigen response
-​ Follicular (in spleen): cycle through the B cell rich lympohoid follicles
B1 cells: found in the peritoneal and pleural cavities (liver, intestines)
-​ Absent from lymph nodes and blood
-​ Abundant in early development
-​ Fetus: present in the liver as hematopoietic stem cells to start -> organisms begin
dividing on their own -> B2 cells come to dominance
-​ Innate immune response to infection -> DONT require T cell help for activation (no
memory cells)
-​ Activation=spontaneous
Marginal Zone B cells (in spleen: white blood cells- immune cells)
-​ TIndependent and TDependent antibody responses
-​ Activated with+without T cells faster than conventional B cells
-​ Start producing antibodies quickly -> against polysaccharides of blood-borne bacteria

Development (fetus: liver, adult: bone marrow)

-​ Bone Marrow- Adult: hematopoietic cells -> immature B cells: via Ig Gene
Rearrangement VJ + VDJ recombination (freeze cells at the developmental stage -> once
frozen=> die b/c of non-reproductive rearrangement)
-​ B cells will express IgM+ on surface
-​ Immature cells migrates to SPLEEN to complete
-​ Fully mature B cells: express IgM AND IgD on surface w/ same specificity
 -​ Stay in spleen, move to different organs
-​ Differentiation process: turn into plasma cells or memory B cells with
class switching and affinity maturation

-​ Signals come from STROMA: provides receptor -> receptor interactions and cytokines
and hemokines to guide the process of hematopoiesis
-​ Hematopoietic cells: stay close to bone marrow -> based on the different signals
they receive, they lineage into their respective places
Early ProB: lymphoid stem
cells

ProB cell stage: VDJ

recombination with heavy
chain recombining first D ->
J with the help of Rad
Proteins and TDT (enzyme)
- B2-20: marker expressed
in mouse to follow B cells
- (on B220) Ig α and Ig β:
hemoglobulin that
molecules use to get to the
cell surface and signaling
(there is little intercellular
surface)
- TDT and RAG1/RAG2 present; no presence of H/L chains
 - Pre-B cell stage: VDJ recombination w/ VDJ fused and TDT adding extra nucleotides
​ - RAG1/RAG2, TDT, H/L chains present (surrogate L chain)
- surrogate: replaces a light chain temporarily just to help cell get to surface
(chaperoning)
- Immature B cell: RAG1/RAG2 present, H/real L chains present, I; NO TDT
- Mature B cell: NO RAG1/RAG2, NO TDT (will mess up rearrangement; act as receptors)
​ ​ - H chain: μ+δ, L chain: λ/ ƙ
​ ​ - express IgM and IgD on the surface; populate the secondary lymphoid organs ​
​ - Activated B cell: class switching, somatic mutation, antigen stimulation (happens in
peripheral lymphoid tissue)
-​ Low levels of Ig- IgM, IgD, IgG, IgA, IgE: present (can be short/long-lived)
​ -Plasma Cell: secretes high levels of IgM, IgD, IgG, IgA, or IgE: NO RAG1/RAG2, NO TDT,
NO H CHAIN
-​ NO CLASS SWITCHING OR SOMATIC MUTATION
-​ Some will become memory B cells
KNOCKOUT GENES:
- To determine the B cell developmental pathway: KNOCK OUT mice with RAG1 and RAG2
​ - Naked mouse: defect in the genes- FOXN1 gene -/- (absence of hair follicles and
thymus, NO T CELLS)
- BCR and TCR remain rearranged; B/T cell development = arrested
- Ig α and Ig β gene KO: both are heterodimers that have an immunoreceptor tyrosine-based
activation motif
​ - Are w/ heavy chain
- BCR activation -> phosphorylation of ITAMS w/ Src-family kinases
- Those without these genes don’t have a surface Ig and arrest in B cell development
(no effect on T cells) hgf

Disruption of Surrogate Light Chains: heavy chain won’t get to the surface (no expression of
surface Ig) -> no B cell

- comprised of Vpre-B and Y5 -> bind to heavy chain but

DON'T engage with it (conserve portions of H chain on
constant region)
​ - they DON'T have variability
​ - help H chain to get to the surface -> B cells will divide and make copies of themselves
-> individual L-chain rearrangement -> increased repertoire

Allelic Exclusion: H chain -> H chain expression -> inhibits H chain rearrangement ​
- H chain on the surface -> induces
allelic exclusion and initiates L
chain rearrangement
- Human Gene vs. Murine (mouse):
Transgenic: all B cells carry human
b/c there is a presence of
transmembrane domain ->
exclusion of mouse genes

Productive -> non-productive

Bone Marrow:
Negative selection:
-​ antibody for MHC Kk ->
introduced in H2d and H2d/k mice
-​ Immature B cell:
encounters self-antigen within the
bone marrow -> they are eliminated by apoptosis
-​ Deletion: changes the specificity of BCR through receptor editing of the light
chain
Receptor Editing:
-​ Additional rearrangements of L chain -> express BCR with different specificity
-​ Need continued expression of RAG enzymes
-​ 1st rearrangement: between V3 and J3= productive but a combo of light chain with Ab
heavy chain -> autoimmune antibody
 -​ 2nd rearrangement: between V2 and J4= productive but a combo with Ab heavy chain ->
non autoimmune antibody

PART 2:

TIndependent:
-​ B-1 and MZ cells generate these responses to the polysaccharides of blood-borne
bacteria
TIndependent-1 Response:
-​ B cell receives 2 signals: TI-1 antigen and TLR (polyclonal activation that is not
antigen restricted)
-​ Cell wall products (LPS) trigger B cell activation
TIndependent-2 Response:
-​ Antibodies that serve as signaling molecules get engaged -> signal B cells to
activate
-​ Antigen restricted with specific responses -> no memory cells are formed
TDependent:
 1.​ Ig and antigen cross-link (SIGNAL 1) -> increase in MHC II expression
2.​ Ag-Ab complexes= are internalized and degraded to peptides
3.​ Naive T cells DONT have CD40L expression (only happens with T cells are activated)
4.​ Once activated, CD40L is expressed -> interacts with CD40 (SIGNAL 2) -> goes to B cell
5.​ B cell expresses receptors for cytokines -> bound
6.​ Cytokines release TH cells -> B cells are activated, proliferate, and differentiate
7.​ Induces the formation of germinal centers in lymph nodes and spleen

Activated B cells -> B-cell signaling: changes in gene expression, differentiation -> antigens are
processed for presentation on MHC II (B cells act as APC)
-​ B cell co-receptors: CD8 and CD4 -> binding to MHC -> signals to T cells
-​ CD21 (CR2)= complement receptor that binds to C3D -> signaling molecules
-​ CD19= receptor that binds B cells in humans
-​ CD81 (TAPA-1)
-​ Cells= activated in lymphoid organs: blood-borne=spleen, tissue spaces=lymph nodes

SPLEEN: marginal zone B cells -> make TI responses to blood-borne antigens

-​ MZ B: capture and present Ag to T cells faster, secrete IgM
-​ Help with CD1-dependent glycolipid presentation to natural killer T cells
LYMPH NODE: TD activation of B cells
1.​ Antigen enters through afferent lymphatic vessels
2.​ Naive B cells migrate to the T cell border, where there is a recognition of antigens
-​ B-cell activation
-​ Vessels enter and activate the CXCR5: receptor for CXCL13 (a hemokine that is
expressed on naive B cells)
-​ B cells increase expression of CCR7, a receptor that produces T zone chemokines
CCL19 and CCL21 (expressed in T cell zone) -> B cells will migrate to T cell zone
3.​ Antigen-presenting B cells interact with TH cells at the B/T cell border
4.​ Activated B and TH cells migrate -> form primary follicle
5.​ Primary follicles -> secondary follicles with germinal centers -> somatic mutation, affinity
maturation, class switching, form plasma cells -> plasma cells secrete antibodies ->
excrete CXCR4: binds to CXCL12
Germinal center:
-​ Activated B cells w/o antibodies (centroblasts) -> divide and undergo somatic
hypermutation: VDJ region (CDRs that interact with antigen) -> make B cells with high
affinity -> cells express centrocytes and high affinity clones= compete with each other
with FDC-bound antigen (selection: figure out which to preserve)
Follicular dendritic cells: (not blood-derived cells)
 - Follicular dendritic cells: (not
blood-derived cells) have no dendrites
​ - Highest affinity receptors:
dendrites that have accumulated
antigens that were captured on the
surface of the cell: iccosomes; these
were opsonized by antibodies -> FDC
have FC gamma receptors that
opsonize/complement receptors that
breakdown products

-​ B cells migrate with FDC b/c of the binding of CXCL13 with the CXCR5 receptor
-​ FDCs synthesize CXCL13
-​ Only cells with high affinity for cells will survive, and B cells will pull off the membrane
from the FDC
-​ GC B cells compete for T cell help with B cells with high-affinity receptors

-Class switching: requires

CD40/CD40L interactions
-cytokines determine the
isotype produced
-Memory: antibody produced
in secondary response: IgG,
IgA, and IgE (these express high
levels of MHC II and antibodies
of higher avg affinity)
- Memory B cells -> become
plasma cells: direct all energy
to antibody production
- Plasma Cells:
​ - Short-lived: MZ, B1, and non-GC B2 B cells -> make IgM
​ - Long-lived: B2 GC derived

-​ Under the epithelial layer -> Peyer’s

patches: have large numbers of B cells,
plasma cells, and TH cells
-​ Site of IgA production
-​ M cells: endocytose Ag in the lumen -> allow selective passage of antigens through the
body -> allow B cells to make antibodies

Poly-Ig receptor: constant region of IgA

-​ Antibodies are released in the lumen in the gut
and pumps IgA
-​ Antigens are transported by M cells to activate B
cells in follicle
-​ Plasma cells migrate to submucosa and secrete
IgA
T/B Collaboration
B cells will pick up an antigen through their immunoglobulin on the cell surface -> bring in and
present epitopes on the cell surface -> will be displayed: the epitope will be hooked onto the
cell that it sees
-​ T cells will not see the same epitope
-​ Signal 1: B cell continues to express CD40 -> induces B7 on the B cell -> activiates
interaction with TH cell -> makes CD4L which bind CD40 -> signal 2
-​ Signal 2: B cells express cytokines -> B cells can proliferate and differentiate into
plasma/memory cells
Haptens: bind an antibody and NEEDS a carrier molecule to induce a response
-​ Carrier effect: Hapten must be linked to the carrier to elicit a response
-​ To receive a recall (memory) response, secondary immunization must be with same
hapten carrier conjugate
-​ 2ndary anti-DNP plaque forming assay: looking for recall response where red blood cells
that are conjugated to Di-nitrophenol (not a peptide)
-​ Make a lawn of red blood cells -> coat with DNP (hapten) -> dump B cells on top
-> if B cells make antibodies against DNP -> lyse red cells -> formation of plaques
= # of B cells that make DNP

-​ Individual hapten + BSA (not physically linked): 2nd immunication with same -> NO
RECALL RESPONSE
-​ Hapten linked in the 1st immunization 1st week, 2nd week unlinked hapten carrier -> NO
RECALL RESPONSE
-​ Hapten carrier conjugate immunized w/ same hapten but a different carrier -> NO
RECALL RESPONSE
-​ Hpaten carrier conjugate with a newly introduced protein -> 2nd immunization with new
carrier -> YES recall response
Physically linked:
Memory B cells to the DNP, no memory T cells

Recall response to BOTH

Unlinked -> no memory B cells (no protein

associated to it) and no memory T cells

Do NOT get recal antibodies b/c there are no

memory B cells to the DNP

Will get recal antibodies to BSA b/c it is

immunogenic

Hapten carrier conjugate

Memory B cells to DNP and BSA but different carrier

No memory B/T cells to BGG -> no recall response

No second carrier to DNP -> no recall response

Hapten carrier conjugate + 2nd carrier (not

physically linked)

Hapten = physically linked to carrier-> memory

cells -> recall responses for DNP and BGG
-​ w/o T cell help to carrier -> for hapten to
work
In Vivo Injection:
-​ OVA: chicken protein
-​ DNP-BGG: mouse protein
-​ Different antigen complexes:
-​ DNP-KLH and DNP-KNP + OVA: no hapten connection with a carrier that the
animal has seen before -> yes primary response but no recall response
-​ T cells must be specific for that molecule
-​ Antigen bridges the t cell and hapten
In Vitro Injection:
Transpo plate: tissue culture well
that allows fluids to go through but
not cells (cells added in the
presence of APCs)
-​ 1. No antigens
-​ 2. Yes recall antigen
response: carrier will make
cytokines and filter through to
effect T cells

**Bone Marrow Chimeras:

-​ B MHC = not restricted, B cell response is MHC restricted
-​ Mouse strain = transplanted from the bone marrow of a different mouse -> mouse = die
from infection
 Bone marrow of mouse of SAME
strain -> mouse = live

MHC II on APCs must be

compatible

Matching haplotypes => mouse will

LIVE

BM from B: bone marrow makes T cells,

B cells, KLH cells, and dendritic cells
-​ Will generate APCs -> T cells will
go to the thymus and undergo
pos/negative selection (pos: in thymic
epithelial) -> to recognize host strain = B
-​ T cells are B strain restricted (rec
B haplotype)
-​ KLH-specific T cell -> can bind to
-​ B cells will bind TNP (epitope)
and make antibodies

BM from A: APCs come from bone marrow -> T cells go into the thymus in the thymic epithelial
cells
-​ A strain T cells restricted to B haplotype
-​ Genetically A cells but think they’re B cells -> T cell receptor will not react with MHC
molecule -> will not make cells b/c MHC cells do not match the thymus

MHC Restriction: requires recognition of TCR on both the peptide and flanking regions of MHC
-​ CLT will only kill infected cells of the correct self-haplotype
-​ Specificfity is defined by both peptide AND MHC
-​ Locking mechanisms between the two
Dual Reactive Hybridoma:
TCR hybridomas: react with 2 different Cyt C molecules with the same MHC restriction (only
difference: they are off by 1 amino acid each)
-​ Mutated residues in MHC II -> the more IL2 it made, the more affinity the TCR receptor
would be for the MHC complex
-​ Different residues on MHC complex -> different patterns with different AA of MHC II (still
binds to the MHC but with different reactivity and affinity)
-​ If it mutates AA in TCR Valpha or Vbeta: similar changes
-​ TCR and MHC peptide = fluid -> different conformations and affinity

Alloreactivity:
1. Minor component
2. Direct binding -> allows a direct
attack as soon as the APC is not
recognized

Cellular Immune Response to MCH with

allogenic cells -> seen in vitro
-​ Mix 2 cell types together -> react
-> mimic graft rejection -> cell
proliferation -> cell killing

Mixed Leukocyte Reaction MLR: stimulator cells are irradiated and responder cells = not, will
only see reaction of responder cells and the bigger the MHC mismatch = the stronger the
response

Mix between responder and stimulator

cells
-​ All responder cells are B
-​ Stimualtor cells have
mismatchees: more mismatches->
stronger proliferation
Memory and Cytotoxic T cells
Expansion -> Differentiation-> Elimination of antigens -> Apoptosis -> Memory

1.​ T cell activation and differentiation:

a.​ Signals: 1. TCR receptor on MHC, 2. CD28 response -> effector CD4 T cells and
helper cells

-​ Memory cells must be reactivated to remember the cells

T cell Memory:

Central memory: have ability to reactivated

and divide fast
-​ Need 2ndary exposure
Effector memory: pronounced effector
function -> found in tertiary tissue (ex.
skin): easy to activate and kill infected cells
Resident memory: go to place that is
orginated activated
 MC Arise:

A. Separate precursor model: the cells are already

determined they become memory cells or effector cells
B. Decrease potential model: T cells are exposed to
signaling multiple times -> after multiple signals, it
changes the type of memory cell
C. signal strength: one signal with different levels of
interactions
-​ Stronger interactions: Effector cells
-​ Weak interactions: CM cells
D. asym cell fate model: T cell divides -> daughter cells become an effector cell that interacts
with the memory cell

Maintaining memory cells:

Cytokines: IL7 and IL15 = induce cell division and proliferation
-​ IL7: not based on macrophage
-​ IL15: dendritic cells
CD8: long memory cells
-​ Dont need B cells or Ag:Ab complexes to maintain the CD8 memory
-​ Dont need CD4 cells to maintain CD8 memory
CD4: memory is lost over time
-​ B cells + Ag:Ab complexes are needed to maintain
B cell memory: response = faster
-​ Undergone somatic hypermutation -> exposed to antigen already
Cytotoxic Response:
-​ T-cell mediated: CD8 cells -> function: kill infected cells (tumor cells) : destroy cells and
reject allografts
-​ MHC restriction for T cells: T cell receptor recognizes the MHC
-​ In the thymus: positive selection

Depending on the 1 amino acid

that you change, you can
increase affinity to TCR in the
thymus

T cells that receive only 1 signal from receptor-> get anergic

-​ Need second signal to get energized by dendritic cells

Graft rejection: non-APC cells will induce anergy rather than activation b/c of the lack of
co-stimulatory molecules
-​ Activated APCs w/ graft -> host the CD4 and CD8 T cells
-​ Donor vasculature= destroyed and the graft dies
 No second signal (not a dendritic cell)

Killing cells by CTLs: T cells have

granules (with cathespin B) that
release perforin (C6,7,8,9) that creates
pores that poke holes in the
membrane and release proteases ->
degrades the cellular contents
-​ Doesnt kill the CTL: cathespin B
binds to the CTL membrane and
degrades peforin but protects Ctl

CD8 from perforin knockout mice have CTL activity

- FAS- L: binds to FAS expressed on target cell -> expression is increased on cells -> Fas + Fas-L
interaction-> causes apoptotic death of target cell
​ - for CTL to bind to target cells -> TCR must engage with MHC and peptide
Tolerance: specific negative immunity
-​ Mechanisms:
-​ 1. Clonal deletion: delete of Ag-specific cells -> in primary lymphoid tissues (bone
marrow, thymus)
-​ 2. Clonal anergy: clone is present but cant response
-​ 3. Clonal suppression: response re-appears if suppression is removed
Clonal deletion (negative selection):
-​ Irreversible
-​ VB17 T cell receptor model -> cells are not expressed on peripheral T cells in mice that
have MCH II E (expressed on cortical thymocytes: immature)
-​ If strain has E on immature thymos will express VB17 on the surface
-​ If strain has on mature thymos: low levels of T receptor on the surface
-​ Bone marrow that expresses E -> no T cells on the surface
-​ Host dendritic cells dies from irradiation -> replaced by donor cells from donor
marrow
-​ Negative selection occurs on donor-derived donor cells
Clonal anergy: Ag signals T cell -> cell turns off and on (signal 1: TCR, signal 2: co-stimulation)
-​ CD28 bind to B7 complexes (co-stimualtion)
-​ IL1 co-stimulates to TH2 cells
Clonal deletion: in B cells -> B cells get eliminated if specific for cell surface self Ag seen in B

Peripheral deletion of B cells: B cells = eliminated in periphery NOT BM

-​ Expressed in liver
-​ b/c CD4 is activated to help
-​ CD40 on the B cell needs to be
ligated to CD40L on the activated T cells
or the B cell dies
-​
Self-souble Antigen- B cell tolerance:
Tgenic mouse 1 crossed with tgenic mouse 2 -> makes F-1 and secretees HEL with high levels of
soluble self Ag -> causes alternation of b cell phenotypes

Supression :
CD4 T cells = suppress immune responses
-​ Express cytokines with angeric
abilities suppress non-specific
responses
-​ If T reg cells are remoned, the
Ag-reactivity ensues
-​ Temporary tolerance mechanism

Initiation of Proper Immune Response:

1.​ TCR and Ag binding
2.​ Co-stimualtaory signal for both B and T cells
3.​ Stress from damaged cells -> activiates APCs and initiates inflammation

Autoimmunity:
-​ Organ specific disease: response targets an organ with direct cellular damage
Tissuse function is stimulated or
blocked by Abs
Hashimoto’s Thyroiditis: DTH
(delayed type hypersentsitivity)
response to thyroid Ags

Pernicious Anemia: make auto

antibodies that bounds an intrinsic
factors -> blocks vitamin B12 -> Ab
block -> hematopoisesis is altered -> anemic

Autoimmune Hemolytic Anemia: autoAb that are directed against red blood cells ->
intereaction with antibodies and RBC -> complement lysis of RBC

Insulin-dependent Diabetes mellitis: DTH + autoAb against the b islet cells in pancreas ->
destory islete cells and issulin production is reduced.
Graves disease: autoAb stimualte
hormone R -> thyroid hormones
produce

Myasthenia Gravis: blocks Ab to

acetylcholine R -> inhibits musicale
activation

-​ Systemic disease: response that direceted against a range of Ags

-​ Involves multiple organs and tissues

Multiple Sclerosis: reactive T cells

and they attack protein fibers

SLE: multiple Ab to DNA, histones,

… -> immune complexes create and
destory the walls of Blos -> rashes
and sun sensitivity

Rhematoid arthritis: IgG that is

anti-IgG Fe -> localize to joints that
activate C’ and inflammation occurs

Ankylosing spondyltisis: HLA-B27

(class 1 molecules) that destroy the
large joints
Thymic Selection
Peptide complexes with TCR that recongize MHC molecules
Bone Marrow Chimera: bone marrow from red mouse to white mouse
-​ In thymus: APCs are selected on thymic epithelial -> negative selection: fails positive
selection, all pass negative selection with repertoire = skewed
-​ CD4 cells are restricted to host MHC and APC are on donor MHC
Antibody Mediated Hypersensitivity Reactions

Hypersensitivity:
Type 1:
Immune mediator: IgE -> bound to mast cells and basophils -> release of vasoactive mediators
-> asthma, allergies, eczema, edema
-​ 1. Primary exposure to Allergen: allergen is carried to lymph nodes -> B cell with
ag-specifc is activated with help of T cell > class switching produces IIgE
-​ Fc region of IgE binds to FCER1 on mast cells and basophils -> sensitizing the cells
-​ 2. Rapid of response: Allergen binds to FCER1 with IgE on the primed mast cells ->
receptor cross-linking -> causes degranulation of mast cells
-​ Mediators are released -> symptoms: rash, itching, pain
-​ IN basophil -> there is crosslinking of the receptors that triggers the receptor mediated
activation and degranulation
 Densensitization therapy (hyposensitization):
-​ Immunotherapy with repeated injection of increasing doses of allergens -> shift from IgE
to IgG acting as a block to Ab
-​ IgG: completing and triggering FCYRIIB

-​ Environmental factors and genetics -> influence allergies

-​ Air pollution, low fiber diet, farm enviroment
-​ Allergen sensitization through skin: as the allergn goes through the skin -> loss of
epithelial integrity
-​ IL4: regulates IgE
-​ IL4 and IL13: overlapping activity -> those have have deletions of either have a
decreased level of IgE
-​ LPS: polycolonal activator of B cells
Different cytokines induce
transcription from different 1
regions.
- IL4 signaling activate
transctiption factors that bind
to promotor IE and initiate
transcription
-> how to know?: take B cells
and stimulate them on
cytokine, use BCR to look at
which type of messaging is
there
 Want open chromatin -> need
AID to bring enzyme in
- CD40/CD40L : induce
expression of AID -> triggers
DNA rearrangement

IgE: help play a role in

immune response against
intestinal helminth infections
(worm)
- eosinophils

Type 2:
Immune mediator: IgG or IgM -> Ab on cell surface mediates cell destruction with complement
activation and ADCC (ab-dependent cell mediated cytotoxicity) -> poke holes in the membrane
-> blood transfusion reaction, anemia
-​ Activation of complement system -> formation of MAC
-​ Reactions: 1. Transfusion rxn: donor RBC = not matched to blood type -> activation of
complement and cell lysis
-​ 2. Hemolytic disease of newborn: maternal IgG speciic for fetal blood-group Ags
crosses placenta and destroys fetal RBCs
 Mismatching of blood groups:
- RH -: mother, RH +: father -> baby:
RH + -> RH + will be expressed on cell
surface and fetal blood with mix with
materaln blood -> production of IgG

- Next child: RH + -> Ab cross and

encounter memory cells -> make IgG
and it crosses the placenta and binds
to RBC of fetus -> cause complement
mediated lysis of fetal RBC and
releasing hemoglobin -> damage
brain
Prevention: injection of rhogan
-​ Injection of IgG Abs that bind to the RH antigen on fetal RBC to prevent sensitization of
mother

Type 3:
Immune mediator: immune complexes -> Ag-Ab complexes are deposited in tissues -> ensues
an inflammatory response by infiltration of neutrophils -> arthus reaction and serum sickness
(vasculitis, lupus, rheumatoid arthritis)

-​
Type 4:
Immune mediator: T cells (TH1, TH2) release cytokines -> activate macrophages of TC cells ->
direct damage -> graft rejection, dermatitis
Immunodeficieny Disorders:
-​ Primary: inherited genetic or developmental defect in system
-​ Occurs in the Humoral B cells
-​ APECED/IPEX: mutations in error -> not tolerant
-​ Bare-lymphocyte syndrome: defect in promoter in class 2 -> NO CLASS 2
EXPRESSION -> reduced CD4 help
-​ Hyper IgM syndrome

-​
Stem Cell Gene Therapy for SCID Gene Defect:
-​ Gene from gamma -> goes into stem cell and integrates -> reverse transcribe -> replaces
gamma chain -> permanent change in stem cells
-​ Stem cell source: moblize cells in blood -> add GCSF or AMD3400 -> counteracts
the CXC4: that holds onto the marrow
-​ CD34: can remove the cells and add cytokines -> give back to patient
-​ Therapeutic gene = insereted near the LMO2 proto-oncogene -> which is typically
expressed in HSCs not T cells
-​ Increased LMO2 expression= increased replication

Secondary: acquire -> loss of immune function that results from exposure to an external agent
HIV:
-​ 2 VIRAL proteins:
-​ T M : transemebrane
-​ SU: surface molecule -> binds to receptor CD4 to virus and non-covalently
attached to T M (detergent will remove)
-​ Inside core = 2 ssRNA + viral enzymes:
-​ Integrase: viral encoded that causes integration-> takes RNA to reverse
transcribes into DNA
-​ Protease: cleaves precursor proteins in the virus during differentiation
-​ If virus buds out, proteases cleaves proteins that allows core to
fold
-​ Genomonic structure:
-​ LTR: long terminal repeat @both ends -> promoter
-​ GAG: encodes code proteins that make coffin
-​ POL: encodes enzymes
-​ ENV: encodes T M and SU
-​ OBILIGATE genes:
-​ TAT: transactivator -> virus wont replicate w/o it
-​ REV: affects splicing patterns -> controls reading frames
-​ Virus will not replicate w/o it
-​ VPF: allows RNA/DNA genome to transcribe and pass through the nuclear pore
-​ VIF/VPU: counteract cellular factors as antivirals -> bypass cells to get rid of
infection
-​ NEF: downregulates CD4 -> prevents superinfection + cell stays alive a little
longer
-​ If CD4 is removed, the cell membrane’s affinity decreases, and the cell
can lyse faster
1. Enemy binds to SU and binds to
CD4
2. Conformational change -> exposes
T M that can bind to coreceptors:
CXCR4 and CCR5: chemokine
receptors
3. Conformational change once
bound -> T M fuses into the cell
membrane
 4. Enzyme reverse transcriptase encoded by the virus causes ssRNA molecules to
become 1 dsDNA
5. Complex goes through the nuclear pore with the cell not divided (macrophages)
6. Once in, the integrase protein integrates genome into chromosome -> viral gene
transcription and translation on surface
7. Protease process occurs just as it buds out

CXCR4 and CCR5: coreceptors for HIV infection

-​ Some bind both -> but dont need both at the same time
-​ Inhibit the binding of coreceptors
-​ CCR5: RANTES, MIP1A, MIP1B-> bind and inhibit entry
Once Virus gets in:
-​ Virus gets on cell surface -> gets picked up by dendritic cell (Langerhans cell) -> brings
inc contact with CD4 -> contact

Starts 1200
-​ Fall without treatment,
rebound, sympotmless phase, fall
with AIDs and death
-​ CTL are killing target cells
Immune Response to HIV:
-​ Ab to viral proteins are made but they are poor neutralizing (can’t target cells well b/c
epitopes are hidden)
-​ Viral env proteins mutates and changes quickly -> escapes from Ab response
-​ CD4 respond but # are low
-​ CD8 = present -> 50% are assessed
-​ NEF downreg MHC I HL1A-A and -B (not C) -> decreased CD8 mediated killing
-​ APC function = pertubed -> demand to replace lost T cells that alter BM function

HIV reverse transcriptase= error prone

-​ Mutations: drug resisitance, alterd tropism, increase replication potential
-​ In AIDS -> deletion of CD4+ T cells in GI tract
HIV persistence:
-​ Short-lived infected cells + long-lived infected cells -> antiretroviral therapy attacks
short-lived cells but nothing to long-lived -> creates reservoirs
-​ If you go off therapy -> the cells come back
-​ Activated T cells die in about 24 hrs but come back
-​ HIV enters the cell by binding to CD4
-​ CCR5 delta 32: a mutation where CCR5 is not functional -> resistant to infection
with most strains of HIV
-​ Berlin Patient: 2 bone marrow transplants from CCR5 delta 32 ->
graft-host reaction (reverse: graft rejected host)
-​ New immune cells were not susceptible to HIV
-​ Targeting the latent reservoir:
-​ Make latent cells visible to target and kill
-​ Immunotoxin: Ab that is linked to toxic molecule -> antibody will bind to virus
proteins on cell which cause toxin to kill the cell before the new virus is made
-​ Could eliminate latent infected cells so patients stop medication
-​ Experiment: adoptive transfer of NK cells -> delay viral rebound in humanized mouse
model of HIV-1 latency
-​ Mice with NK cells -> had less HIV DNA and barcode diversity compared to
control mice
-​ Mice with LRA and NK cells had delayed viral rebound and decreased barcode
diversity
-​ Kick and Kill: reduce/eliminate the viral reservoir
Infectious Diseases and Vaccines:
-​ Chemical + physical barriers = have to be breached to cause infection
-​ Epithelial linings of skin and gut = barriers
-​ Breaching pathogens -> encounters innate mechanisms -> leads to recruitment
and initiation of adaptive mechanisms
-​ Pathogens: clear or kill the host
-​ Viral infections: enter cells through cell surface receptors
-​ Replication occurs inside -> viruses thrive in if they don’t kill the host
-​ Viruses trigger humoral and cell-mediated adaptive immunity
mechanisms

Influenza:
-​ A, B, C
-​ Glycoproteins:
-​ Hemagglutinin HA: allows attachment of virus to cells
-​ Neuraminidase NA: helps new virus escape from host cells
-​ Ag drift: high mutation potential of RNA genome -> RNA polymerase lacks proofreading
capability
-​ Ag shift: different virus types infect a single cell -> create new HA/NA combinations ->
pandemics

Bacteria evades host defense

mechanisms at several diferent stages
- attachment to host cells: bloclage of
attachment by secretory IgA Ab
- proliferation: phagocytosis +
complement-mediated lysis and
localized inflammatory response
- invasion of host tissues: Ab mediated aggulation
- toxin-induced damage to host cells: neutralization of toxin by antibody

IgE response -> will bind to

basophils
Worms: innate immune response

Commensal organisms: crowd out ->

get rid of normal flora
(non-pathogenic=good for you)

Emerging vs. Re-emerging infections:

-​ Emerging: something new not observed (HIV, COVID)
 -​ Re-emerging: something old and coming back again (TB)
-​ Due to globalization and interaction between people and wildlife
SARS-COV-2:
-​ Binds to the receptor ACE 2 and the TMPRSS2 protease
-​ Tissues with high levels of ACE-2 R expression: in lungs, small intestine, heart,
kidneys
-​ Activates the spike protein and enters the cell
-​ In Aleveolus -> type 1: brings oxygen in, and type 2: releases liquid that swells the
edema, fluids come in-> collapses and can’t do gas exchange -> more inflammation ->
cells are destroyed
-​ Diseases may re-emerge for reasons: a combination of diseases (HIV and TB)
-​ Laxity in vaccination program adherence
Vaccines:
Active immuniziation to induce
immunity and memory
-​ Small measle outbreaks due
to drop in herd immunity levels
(due to lax vaccination adherence)
Conjugate/ multivalent vaccines = improve immunogenicity and outcome
-​ Molecules arent strong enough Ag on their own to stimulate a good response
-​ Simple linking of weak Ag with a stronger one
-​ Good at Ab induction but to induce a CTL response -> deliver Ag into cells in MHC
class 1 molecules
-​ Creation of lipid carries = immunostimulating complexes ISCOMs
RNA based Vaccines:
-​ Contain mRNA, lipid nanoparticles to protect RNA, salts, and sucrose

-​
Cancer and the Immune System

Mutations: induce malignant transformation -> induced by Carcinogenic agents

-​ Cellular genes are associated with cancer formation:
-​ Oncogenes: activate growth and proliferation -> promote cancer when activity
enhanced
-​ Tumor supressor genes: inhibit cell proliferation and mediate DNA repair ->
promote cancer when activity is lost
-​ Apoptosis gene: controls programmed cell death

Tumor antigens: TSA (tumor specific Ag) vs. TAA (tumor associated Ag):
-​ TSA: non-self antigens that are expressed in tumor
-​ Mutations in tumor cell -> neoantigens
-​ Antigens derived from viral proteins -> HPV and cervical cancer
-​ TAA: wild type protein that display abnormal patterns
-​ Oncofetal tumor Ag: proteins that are expressed during embryonic development
and absent in adults -> gets re-expressed in the tumor: CEA and AFP
-​ Antigens expressed at low levels -> growth factors like EGFR
-​ Are recognized by immune system
 Immune system startts
to get suppressed with
non-active immune and
dendritic cells

Tumor gets mutations

that allow it to grow

Negative aspects:
-​ Chronic inflammation -> promotes cancer
-​ Increases cellular stress signals -> genotoxic stress -> greater tumor cell invasion
-​ Enhanced production of anti-tumor antibodies -> free tumor Ag binds to Fc receptors on
NK cells and macrophages -> impairs their anti-tumor activities and blocks cell surface
Ag
-​ Active immunosuppression in tumor microenvironments: releases soluble factors that
suppress immunity (TGF-B, indoleamine 2,3 dioxygenase IDO, IL10)
-​ T reg cells and MDSCs -> increase M2-macrophases, TH2, and TH17 populations
immune response to cancer:

Cancer treatments:
1. Surgery
2. Raditation
3. Drug therapies: chemotherapy aimed at blocking DNA synthesis and cell division
-​ Hormonal therapies -> interfere with tumor-cell growth
-​ Targeted therapies with small-molecule inhibitors of cancer
-​ Immunotherapies -> induce/enhance the anti-tumor immune response

1. Monoclonal antibodies: see antigen and

bind to it -> express cytotoxic response
2. Immune checkpoint: block the antigen
Bispecific antibody: bind to T cells -> activate the response
Target cell that expressed the antigen ->
antigen that expresses the domain and
binds to CD3 -> activiates the T cell and
lysozome -> kill the cell

Antibodies that are conjugated to toxins: conjugate to chemotherapy -> good results
-​ Coinhibiotry receptors: CTLA-4 and PD1 -> promote T cell anergy and exhaustion
-​ CD28: promote native T cell activation

-​
Proteins: ANTI- CDL4 + ANTI-PD1: block interaction
-​ Anti-CDL4: binding partner- CD86 (expressed on antigen presenting cells)
-​ CDL4 blocks the induction of immune response
-​ Tumor is releasing antigens -> antigens is picked up by dendritic cells and
go to lymph nodes
-​ If T Cells are releasing CDL4 -> when antigen-presenting cell presents
antigen on +1 and +2 -> presents on +2 and binds on CD4 T cell: if T cell
expresses the CDL4 instead, when it binds to the ligand it binds to CD86
 -> provide co-inhibition and stops proliferation of CD4 T cells -> no
functioning CD8 cells -> go to tumor and start killing
-​ PDL1: binds to PD1 and inhibits T cell

Therapeutic Cancer Vaccines: Enhance the anti-tumor immune response

-​ Patient DC= cultured with prostate cancer Ag adn prolif/activatio of cytokines
-​ Primed DC = reinfused, which provokes a CTL response
Neoantigen specific therapeutic vaccines -> enhance the anti-tumor immune response -> use
peptide pools or mRNA strings