Brown University

BIOL 0530 Fall 2015

Principles of
Immunology

Lecture 3:

Immune
Cells and
Organs
Thursday September 17th, 2015
Richard Bungiro, Ph.D.
Richard_Bungiro@Brown.edu

Chapter 2
Organization
Cells of the Immune System
Primary Lymphoid Organs
Where Immune Cells Develop
Secondary Lymphoid Organs
Where the Immune Response
Is Initiated
2

Eventually, all of this will make

sense
T epitope
(Really, it will)
MHC II TCR
B epitope

Helper
T cell

Activated
Th cell

ANTIGEN
peptide

Antigen
Presenting
Cell (APC)
B Cell

IL-4
Plasma Cell
(effector B)
Secreted
Antibodies

Activated
Th cells

viral
peptide

TCR

MHC I

IL-2

Killer
T cell

IFN-

Activated
Macrophage

*
killing

Virus-infected cell3

Cells of the Immune

System
Immune responses
are mediated by a collection of

different white blood cells (WBCs, also known as

leukocytes) which are continuously generated in the
bone marrow through a process called
hematopoiesis

Lymphocytes (B and T cells) are a subset of

leukocytes which mediate adaptive immune responses; these are generated in the primary lymphoid
organs (bone marrow and thymus) and enter the
bloodstream
Circulating lymphocytes transit through blood vessel
walls and enter secondary lymphoid organs (lymph
nodes, spleen, Peyers patches of the gut, etc.)
Lymphocytes constantly recirculate, conducting
surveillance throughout the body
4

Think About
Your Blood

Blood
Fractions

FreshSpun

Hematocrit = percentage of
RBCs by volume; about 38 in
sample at left

Morphology
of Blood
Blood Cell Counts:
Cells
(per mm3, Table 2-1)

Red Blood Cells

(RBCs or
Erythrocytes) 5.0 x
106
Platelets -> 2.5 x
105
White Blood Cells
(WBCs or
leukocytes)
7.3 x 103 total:
50-70% Neutrophils
20-40% Lymphocytes
1-6% Monocytes

What Do They Really Look

Like?

RED BLOOD PLATELETS LYMPHOCYTE MONOCYTE

CELLS

NEUTROPHILEOSINOPHIL BASOPHIL
GRANULOCYTES

Hematopoies
is

Literally, the making of blood (Greek)

Process of continuously generating blood

cells (RBC, WBC & platelets) from stem cells
which begins shortly after fertilization and
continues throughout lifetime
Hematopoietic Stem Cells: self-renewing cells
that give rise to all other blood cells (about
0.05% of total bone marrow cells in mice)
Progenitor population: committed to a given
lineage, little or no capacity for self-renewal
Terminally differentiated population:
nondividing cells w/ limited lifespan
9

Hematopoiesis (Kuby Fig

2-1)

Various secreted
and membranebound factors
direct
hematopoiesis
(HP)
Stromal cells in
the bone marrow
provide a HPinducing microenvironment
To maintain
steady-state
levels, the HP
system must
produce 3.7 x 1011
white cells per day
HP system is
10

Sorting Out Cell

Populations
Via Differentiation
Distinguishing between
Antigens
various blood cells has
involved the development of
surface-reactive monoclonal
antibodies (mAbs)

Each mAb is classified based

on its reactivity with a
particular cell surface
molecule, populations of which
are expressed in different
patterns as cells develop
These are called cluster of
differentiation (CD) antigens
See Kuby Table 2-3
At least 350 human CD markers
have been described (see
Appendix section of Kuby)

en.wikipedia.org/w
iki/File:Cluster_of_
differentiation.svg

11

12

Search for the

Hematopoietic Stem
Cell

About 10,000 raw (unpurified) bone marrow cells can restore

the hematopoietic system of a lethally irradiated mouse
Weismann et al. used cell sorting techniques to enrich a
preparation of raw bone marrow cells

- They first used antibodies to differentiation antigens (surface

molecules that are expressed on developing blood cells) to

remove mature (lineage positive; Lin +) cells NEGATIVE selection
- They then treated the remaining lineage negative (Lin -) cells with
antibodies to the early differentiation antigens Sca-1 and c-Kit,
and selected the cells that expressed both POSITIVE selection
- 30-100 cells of the enriched preparation were capable of reconstituting a lethally irradiated mouse (see page 31)

Nakauchi et al. refined the enrichment technique to the point

where a single cell could reconstitute an irradiated mouse
Currently mouse HSCs are defined by a Lin -Sca-1+c-Kit+ (LSK)
phenotype
13

Enrichment of Pluripotent Stem Cells

(Kuby Pg. 31)

Note that antibodies

may be employed for
both positive and
negative selection of
cells during the
14

Regulation of
Hematopoiesis
Bone marrow contains various
cell types that produce soluble
and membrane-bound factors
which direct the differentiation
of hematopoietic cells

Osteoblasts
Endothelial cells of blood vessels
Reticular cells
Sympathetic neurons (!)

Compartmentalization in the
bone marrow creates different
microenvironments, leading to
different lineage commitments
among HP cells
T cells & macrophages in the
periphery also produce factors
that affect the process

Kuby Fig. 2-5

15

Hi, Im NASCAR star Ricky Bobby here to

remind you that like me, every developing
blood cell needs a good pit crew. Remember
that in hematopoiesis, if you aint first,
youre last!

16

Dysregulated
Hematopoiesis
May Result in Leukemia

Chronic Lymphocytic LeukemiaNormal Blood Smear

Apoptotic control is lost
Little or no terminal
differentiation occurs

Left panel:
Right panel:

http://pathwiki.pbworks.com/f/1146144287/blood-23.png
https://courses.stu.qmul.ac.uk/smd/kb/pathology/introcoursepics/imag
17

Hematopoiesis: Clinical
Aspects

Blood transfusion (RBCs and platelets)

Regular donation possible because of the

ability of the hematopoietic system to
replace lost cells

Bone marrow transplantation

(especially
with enriched stem cells - see Clinical Focus section,
page 42)

Provide functional immune system in

immunodeficiencies
Replace defective hematopoietic system
Restore the hematopoietic system after
chemotherapy for cancer

Gene therapy w/ transfected stem cells

18

The Inducible Nature of the

Hematopoietic System Makes Regular
Blood Donation Possible

19

A Small Number of
Hematopoietic Stem
Cells Can Give
Someone a New
Immune System
(How Cool Is That?)

http://www.marrow.org/index.html

Dr. B joins the Registry!

20

Lymphocytes
B, T, & Null (NK & NKT) cells

Approximately one trillion in the

human body at steady state
Small lymphocytes (B or T) are
indistinguishable at the light
microscope level
Distinguishable at the molecular
level due to expression of ARMs
(Igs, TCRS) & other differentiation antigens (Table 2-3)
21

Lymphocytes
B cells

Express various forms of immunoglobulin (Ig)

T cells

T Cell Receptor (TCR), CD3, CD4/CD8

NK (Natural Killer) cells

Primitive large granular lymphocytes

Involved in cytotoxic responses against

tumors and certain virally infected cells

Lack Ig, TCR and CD4/CD8

NKT cells

Express receptors typical of NK but also an

invariant TCR
Interact with an MHC-like molecule called CD1
22

The Proliferative Cycle of B

Cells

23

Electron Micrographs of
Lymphocytes
at Various Stages of
Differentiation

Resting

Proliferating

Terminally
Differentiated
24

Monocytes/Macropha
ges
Present in blood as monocytes and in
various tissues as macrophages

Alveolar (lung)
Kuppfer cell (liver)
Microglia (CNS)
Osteoclast (bone)

Monocyte
in blood

Functions include phagocytosis,

antimicrobial activity, tissue repair, and
antigen processing/presentation
Secrete soluble (peptide and nonpeptide) mediators of inflammation

IL-1, IL-6, TNF- , CSFs, prostaglandins

(PGs), leukotrienes (LTs), reactive oxygen
and nitrogen species (e.g. H2O2, HClO-, NO)

25

Monocyte/Macrophage
Macrophages
are 5-10
Morphology
fold larger than the

fold larger
thanFig.
the2-3)
(Kuby
monocytes from which
they develop
Macrophages contain
more organelles than
monocytes, especially
lysosomes
Macrophages have
increased phagocytic
ability
Macrophages may be
activated by various
stimuli such as cytokines
and microbial products
26

Like Bruce Banner, Monocytes

Undergo An Incredible
Transformation

MACROPHAGE
ANGRY!!!
MACROPHAGE

27

Phagocytic
Activity of
Macrophag
es
(Kuby Figure 55)

28

Granulocyt
es

Neutrophils
Also known as polymorphonuclear cells
Phagocytic
(see
Kuby Figure 2-2)
Antimicrobial activity involves various
hydrolytic enzymes (e.g. lysozyme)
Short lived, involved in early
inflammatory responses

Eosinophils
Phagocytic
Involved in anti-parasite defense
Basophils/Mast cells
Non-phagocytic
Bind IgE (but dont produce it)
Release mediators involved in allergy
Mast cells are tissue dwelling
basophilic cells

29

30

Neutrophils (Kuby Fig

2-2a)

31

Eosinophils (Kuby Fig

2-2d)

32

Basophils (Kuby Fig 22b)

33

DC

Dendritic Cells

Professional Antigen
Presenting Cells (APCs)
- High levels of MHC Class II and

costimulatory molecules expressed

Important for activating nave T cells

DCs are present in multiple

tissues as Langerhans DCs,
interstitial DCs, monocytederived DCs, and plasmacytoid
DCs
Follicular dendritic cells (FDCs)
are important in Ag retention
FDCs
and formation of germinal center
reaction in lymph nodes
- Not bone marrow derived, do not
express MHC II

34

The
Lymphoid
System
Consists of primary and
secondary lymphoid
secondary lymphoid
organs

Bone Marrow
Thymus

Sites where
immune
cells are
generated

Spleen
Lymph Nodes
MucosalAssociated
Lymphoid
Tissue (MALT)
Lymphatic
Vessels

Sites
where
immune
cells are
activated

35

Primary Lymphoid
Organs
Sites of foreign Ag-independent lymphocyte

development and self Ag-dependent selection

Bone Marrow (or Bursa of Fabricius in birds)

proB cells -> preB cells -> mature naive B cells
Major events: antibody gene rearrangement &
expression, positive/negative selection
Source of proT cells, which migrate to thymus

Thymus
proT cells migrate to thymic cortex where they
proliferate and develop (Kuby Fig 2-6)
Major events: T Cell Receptor (TCR) gene rearrangement & expression, positive/negative selection
proT cells -> preT -> mature naive T cells
Undergoes age-related atrophy
36

37

Secondary Lymphoid
Organs
Sites of "foreign Agdependent" lymphocyte
activation, proliferation, & differentiation

Lymph Nodes

See Figs. 2-7, 2-8, 2-9

Drain and filter Ag from tissue fluids via lymphatic vessels
B & T cells distributed in cortex & paracortex
Upon Ag exposure, lymphoid follicles -> germinal centers

Spleen

See Fig. 2-10

Filters antigen from the blood
Contains red & white pulp; Periarterial lymphatic sheath
(PALS)
Upon Ag exposure, lymphoid follicles -> germinal centers

Mucosal-Associated Lymphoid Tissue (MALT)

See Figs. 2-11 and 2-12

Located in epithelial mucosa of the gut, repiratory tract, etc.
Tonsils, appendix, Peyers patches of intestine, etc.
There are more antibody-secreting plasma cells in the MALT
than in the spleen, lymph nodes, and bone marrow
combined!

38

Lymphatic Vessels Are Associated With Lymph

Nodes

39

Structure of a Lymph Node (Kuby

Fig 2-8)

40

Structure of the Spleen (Kuby Fig.

2-10)

Note that the spleen is

not supplied by
lymphatic vessels;
antigen enters via the

41

Induction of mucosal
immune responses in the
gut involves specialized
antigen transport-ing M
cells and organized
lymphoid structures
called Peyers patches
(Kuby Figures 2-11 & 2-12)

42

Evolutionary Distribution of Lymphoid Tissues

(Kuby Pg. 58)

Adaptive immunity
evolved in
vertebrates ca 500
mya
Adaptive cells and
molecules arose
through modification
of preexisting innate
components
The earliest structure
to arise was the gutassociated lymphoid
tissue (GALT)
Evolution has added
new immune organs
over time while
retaining the earlier
structures
V(D)J rearrangement
is exclusive to jawed
vertebrates, but the
jawless fishes also
have adaptive
immune systems
43

Hematopoie
sisLymphoi

B cell

II

d
progenit
or cell

Natural Killer
Cells

T cell

Lymphocyte
s
rearrange
DNA to
generate
ARMs

Macrophage
s,Dendritic
Cells

Hematopoieti
c Stem Cell
(HSC) in bone
marrow
(selfrenewing)

Granulocytes
(Neu, Eos, Baso,
Mast)
Myeloid
progenit
or cell

RBCs,
Platele
ts

44

Lymphocyte Development &

Migration
BONE MARROW
HSC proB preB

LYMPH
NODES

proT

BLOOD

SPLEEN

THYMUS
proT

preT

MALT

Primary Lymphoid OrgansSecondary Lymphoid Orga

45

Eventually, all of this will make

sense
T epitope
(Really, it will)
MHC II TCR
B epitope

Helper
T cell

Activated
Th cell

ANTIGEN
peptide

Antigen
Presenting
Cell (APC)
B Cell

IL-4
Plasma Cell
(effector B)
Secreted
Antibodies

Activated
Th cells

viral
peptide

TCR

MHC I

IL-2

Killer
T cell

IFN-

Activated
Macrophage

*
killing

Virus-infected cell
46

47