Immunology 101

Richard T Maziarz, MD
September 13, 2013

Basic concepts
Self vs nonself recognition
Response to danger signals- Inflammation
sterile vs nonsterile

Components of the immune system

Impact of transplantation on immune system.
Immune reconstitution.
Harnessing the immune system

Self vs non-self admission is

evolutionarily conserved

Characteristics of Mamnalian
Immune Response
Self: Nonself Recognition

Protection from foreign pathogens

Elimination of malignant transformed cells
Rejection of foreign allografts
Failure of tolerance--> Autoimmune disorders

MHC molecules
MHC class I
HLA-A,B,C
E, F, G, H

Ubiquitous expression

MHC class II
HLA-DR,DP,DQ
DO,DX

Limited expression

Antigenic Diversity

Crystal structure

MHC Molecules : Diversity

Genetic structure : exon crossover events

Molecular structure : point mutations
Peptide binding pocket
TAP
Protein degradation and processing
End result: Tissue specific antigens
Future: Peptide specific therapies, vaccines?

Immune System
Adaptive immune system vs innate immune
system

Pathogen associated molecular

patterns (PAMPs)
Bacterial components
Viral components
Endogenous sources
Uric acid
DNA
RNA

Toll like receptor activation

Components: Immune System

Lymphocytes (Specificity) : B cells, T cells, NK
cells
Non-Specific Elements
NK Cells
APCs: Monocytes/macrophages; endothelial cells; B
cells; Dendritic cells; Other- microglia, Langerhans
cells, etc
Neutrophils
Soluble mediators: cytokines, lymphokines, chemokines

Effector cells

Immunologic specificity
Antigen-specific receptors on cell surfaces
T lymphocytes: antigen specific T cell receptor
B lymphocytes: surface IgM or IgM + IgD

Antibodies in serum after secretion by

plasma cells

Cluster of Differentiation Antigen

Nomenclature
Nomenclature defines the name of the
antigen : (CD1 CD326)
Developed to clarify the biology of
responses associated with antigen
All CD designations are not necessarily
independent molecules
Examples : CD3, CD11/CD18 are peptides;
CD15 defines CHO sidechain

T cell and B cell clones are

actually individual tissues. The
comlexity of the immune system
is simplified by the recognition
that there are billions of tissues in
circulation contributing to
immune surveillance

T cells
Intrathymic vs Extrathymic differentiation
Thymus differentiation : well ordered pathway cortex--> medulla and out
CD34--> CD34 CD7--> CD2CD7(triple negative)-> CD2CD3 (double negative)--> CD2CD3CD4CD8
(double positive)--> CD2CD3CD4 or CD2CD3CD8

TCR rearrangement accompanies positive and

negative selection

CD4 and CD8 Co-Receptors

Adhesion molecules :
CD4:MHC II
CD8: MHC I

Recognition of MHC - conserved sequences

Co-stimulatory function to augment TCR
signalling (associated with lck)
Defines subpopulation of effector T cells
Helper and cytotoxic T cells

T Cells in the thymus

Positive Selection
Low affinity TCR for self can mature and
migrate to periphery

Negative Selection
High affinity TCR for self are deleted following
TCR ligation

Actual Mechanism remains unclear

Signal intensity vs Developmental stage

Age Related Change in Thymus

Haynes, Ann Rev Imm, 2000

Age Related Change in Thymus

Age

0-1
2-10
11-25
26-49
>50

%TE space
93
88
63
45
18

Thymus out
>109
8.8 x 108
6.3 x 108
4.5 x 108
1.8 x 108

Haynes, Ann Rev Imm, 2000

Costimulation
TCR and CD28 simultaneous activation leads to T cell proliferation
and target cell death
TCR and CD152 (CTLA4) binding can limit a T cell proliferative
response
The lack of cell surface CD80 and CD86 can lead to T cell
programmed cell death or to induction of peripheral tolerance (e.g.
anergy).
Absence of CD80 on tumor can induce tolerance
Heterotypic antibodies binding CD3 and CD28 can overcome
peripheral tolerance
OKT3 treatment in renal transplants can render T cells refractory to
stimulation

 Immunologic Memory:
Antigen exposure--> proliferation and differentiation;
small # of cells may proceed to memory state
Secondary Antigenic Challenge : anamnestic
response

Amplification of Immune Response

Cell: Cell interactions
Cell: soluble mediator interactions

Cytokine Expression by T cell

populations
Nave T cells
IL2, lo induction of DC IL12

Memory T cells
T-CM:(CCR7+):
IL2, induce DC IL12

T-EM:(CCR7-):
lo IL2, IL4, IL5, IFN-G,
Sallusto, Nature, 1999

Memory/Effector T cells
Effector T cells :
Conventional: Precursor of memory cells?
Hypothesis: Downstream cell after
differentiation of T central memory cell?
Supported by telomere shortening observations

Phenotype: CD45RO+CCR7+ and

CD8+CD45RA+CCR7-CD62Llo
subpopulation(?)

Nave T cells: Molecular

Determination
Chromosome 14 is home of alpha and delta
TCR
Rearrangement of TCR alpha locus during
thymic maturation splices out TCR delta locus
T cell receptor excision circles (TREC):
maintained episomally and do not replicate
Antigen driven T cell expansion does not
expand TREC

 Two classes of TCR

TCR are polypeptides with two functions- external
antigen recognition and signal transduction
TCR engage antigen on the cell surface of other cells
TCR recognize antigen in context of MHC
TCR families can be associated with diseases
V2 in toxic shock (super antigen)
V12, 17 in multiple schlerosis (auto antigen)

Generation of TREC

TREC : Clinical Applications

Decreases with age, but persists into 7 th
decade -Douek, Nature, 1999
Decreased in HIV-infected pts
Increased with aggressive antiviral therapy in
HIV
Increased in thymic transplants for DiGeorge
syndrome, coincident with increase
CD45RA+, CD62L+ -Markert, NEJM, 1999

Recovery of CD4 and CD8 T Cells

after AutoTx for MM

CD4= opaque
CD8= open
CD34s=circle

Douek, Lancet 2000

T Cell Receptor Analysis

Normal V- Family Expression

CDR3 Size-Pattern Analysis

Normal

In vitro
Ag stim

HIV

T cell line

Manfras, J Imm Met, 1997

CDR3 Size-Pattern Analysis in

Transplantation
Gorski, JI, 1994:
Repertoire stability
Rate of repertoire recovery
Impact of opportunistic infections

Memon, AAI, 2000:

Correlated with CD4+, CD45RA+ T cell
recovery

TCR signal transduction

Activation of T cell
Render T cells refractory to stimulation
Costimulation (CD3 : CD28)
Targets for therapy of T cell diseases
Genetic defects (SCIDS and others)
Zap 70 deficiency
CD3 gamma or epsilon deletion

Second Signals

T cell sub-subsets
CD4 T cells differentiate into separate
subpopulations of T cell
Th1 are important for cellular responses and resistance
to infection; IL-2, IFN-G, TNF-alpha are produced
Th2 are important for humoral responses and can
increase susceptibility to infection; IL-4, IL-5, Il-6, IL10, IL-13, GM-CSF; can downregulate Th1 responses

CD8 effector cell subpopulations like CD4 cells

have been identified.

Helper T cell subsets

CD4 T cell subsets

Natural killer cells

Killer inhibitory receptors (KIR)

Diversity
HLA recognition
May impact leukemia outcome
Clinical relevance- recognition of herpes
virus family members

Impact of transplantation
conditioning on host immunity
Myeloablative
Immune suppressive
Quantitative and qualitative deficits

Tumor Cell Escape from T cell

Immunosurveillance
Loss of expression of MHC antigens

Transcriptional repression (-) of gene expression

Intracellular competition with 2M
Loss of trans activating + factors for expression
Altered TAP transporter
Chromosomal Deletion

Production of inhibitory cytokines e.g. IL10,

TGF-

Tumor Cell Escape from T cell

Immunosurveillance
Induction of Zeta chain translocation and
anergy
Loss of adhesion molecules
Lack of costimulatory molecules (CD80,
CD86, CD40)
Lack of TSTA, TAA
Secretion of soluble antigen to create tolerance

Immune reconstitution

Population
% Lymphs
Count/ul %
Total T cells(CD3+)
59.8
316 L
CD3+ CD4+
23.4
124 L
CD3+ CD8+
30.9
163 L
CD3+CD4-CD8-(Doub.neg) 4.8
CD3+CD4+8+(Doub. pos)
0.0
CD4+CD45RO- (naive)
4.6
NKT
6.9
NK cells (CD56+CD3-) 38.4 203
Total B-Cells (CD19+) 0.0
0L

Lymphs count/ul
55-82% 800-3500
20-60% 500-2000
15-30% 300-900
NA
<6%
NA
<2%
NA
8-50%
<3%
5-20% 75-500
5-15% 70-450

Immune reconstitution

% of B-cells % of B-cells
IgD+ CD27- Naive B-cells 0.0 L 58-80%
IgD+ CD27+ Non-switched memory B 0.0 L 7-21%
IgG+ CD27+ Switched, memory B 0.0 L 9-19%

Activation Indicators Patient Results

Reference Ranges CD4:CD8 ratio 0.8 L 1.0-3.3
%Lymphs %Lymphs
CD3+CD4+CD8beta+ 0.0 L <1.5%
CD3+HLA-DR+
8.7
<10%
CD4+CD25+ 2.2
<7%
CD8beta+CD69+
1.9 H <1%
% of CD4+ T cells
CD4+CD25+ bright(T regs)
12.0 H <10%

Harnessing the immune system

Antibody therapy
IvIg
BITEs

Cellular therapies
LAK + NK-T
IL2/ cytokine ex vivo expanded cell populations

TIL
Tregs
Dendritic cell vaccines-Provenge
CAR-T cells

Generation of TIL (tumor

infiltrating lymphocytes)

Novel antibody therapy

Hematologic RFS MRD persistence or relapse of ALL

(A) 20 evaluable patients; (B) subset 9 allo HSCT; (C) subset 11 chemo only
T cell engaging, bispecific single chain (BiTE) ab

Topp M S et al. Blood 2012;120:5185-5187

2012 by American Society of Hematology

Chimeric antigen receptor- T

(CAR-T) cells

CAR-T cell applications

CAR-T: mechanism of killing

Clinical Response in the Patient.

Porter DL et al. N Engl J Med 2011;365:725-733.

Expansion and Persistence of Chimeric Antigen Receptor T Cells In Vivo.

Porter DL et al. N Engl J Med 2011;365:725-733.

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