Homework check

Q&A
Any Question from you?

NTTH-HCMIU-IM-2014

Cells of the IS
DIR-page 19-58: SUMMARY
1. Cells of the IS:
Who are they?
Where are they?
Where do they come from (Origin of immune cells )?

NTTH-HCMIU-IM-2014

COMPONENT OF THE IS
CELLS and SOLUBLE MEDIATORS

Fig 1.1 DIR

NTTH-HCMIU-IM-2014

Myeloid
lineage

INNATE

Lymphoid
lineage

ADAPTIVE

Derived from common progenitor cell in bone marrow

- Pluri-potent hematopoietic stem cell
NTTH-HCMIU-IM-2014

CELLS OF INNATE IMMUNITY- Myeloid lineage

1. Phagocytic cells (Phagocytes):
1.1 Mononuclear phagocytes:
Monocytes (circulating cells) and Macrophages (reside in tissues)
1.2 Polymorphonuclear granulocytes (Polymorphs):
with lobed, irregularly shaped- polymorphic- nucleus
2. NK cells, mast cells and platelets

NTTH-HCMIU-IM-2014

CELLS OF INNATE IMMUNITY- PHAGOCYTES

1.1. Monocytes/ Macrophages
-Leukocytes
with
bean-shaped,
horseshoe- shaped or brain-like
convoluted nuclei;
- relatively large (10-18 um in diameter)
- Circulate in blood with half life of 8
hours;
- contain azurophilic granules (dyed
blue)
- contain well- developed Golgi and
many lysosomes.
- Precursors of tissue macrophages
Macrophages: Mononuclear phagocytic cells in tissue; Derive from blood monocytes
How could these cells recognize, adhere and eat the microbial and even tumor cells?
- Scavenger receptors, TLRs, mannose receptors (particular sugar, lipid molecules
on microbes), FcR, C3a/ C5a R: check them later
NTTH-HCMIU-IM-2014

CELLS OF INNATE IMMUNITY- PHAGOCYTES

1.2. Polymorphonuclear leukocytes
Nuclei are multilobed (2 to 5); Granulocytes (Cytoplasmic granules)
Are short-lived (2-3 days), released from BM, 7 million/ min, 10- 20 um.
Based on how the granules are stained with acidic and basic dyes (HE
histological staining) they are classified into:
Neutrophil (PMN, 95% of all
circulating granulocytes): Principal
phagocytic cell of innate immunity, multi
lobed nucleus, granules stained with both
acidic and basic dye (neutral pink)
Eosinophil: Principal defender against
parasites, bi-lobed nucleus, granules
stained with acidic dye such as eosin
(bright red)
Basophil: Functions similar to
eosinophils and mast cells, segmented
nucleus, granules stained with basic dye
such as hematoxylin (dark blue)

NTTH-HCMIU-IM-2014

CELLS OF INNATE IMMUNITY- PHAGOCYTES

1.2. Polymorphonuclear leukocytes (PMNs)
Neutrophil (PMN, 95% of all circulating granulocytes):
Principal phagocytic cell of innate immunity, multi-lobed nucleus,
granules stained with both acidic and basic dye
(neutral pink), 40-75 % leukocytes
Granule type

specific granules (or

"secondary granules")

Protein
Acid hydrolases, muramidase,
myeloperoxidase, bactericidal/permeabilityincreasing
protein (BPI), Defensins, and the serine
proteases neutrophil elastase and cathepsin G
Akaline phosphatase, lysozyme, NADPH
oxidase, collagenase, Lactoferrin ;
Cathelicidin;

tertiary granules

cathepsin and gelatinase

azurophilic granules (or

"primary granules")

Source: Wikipedia with modification

NTTH-HCMIU-IM-2014

CELLS OF INNATE IMMUNITY- PHAGOCYTES

1.2. Polymorphonuclear leukocytes
Eosinophil: Principal defender against parasites, bi-lobed nucleus,
granules stained with acidic dye such as eosin (bright red); 1-6 %
leukocytes
- Granules: Major Basic Protein (MBP), Eosinophil Cationic Protein
(ECP); Eosinophil-Derived Neurotoxin (EDN)
- MBP: potent toxin for helminth worms, induces histamine release
from mast cells, activate neutrophils and platelets and of relevance to
allergy, provokes bronchospasm.
- Release of these molecules is the only way in which eosinophils can
kill large pathogens which cannot be phagocytosed.

NTTH-HCMIU-IM-2014

CELLS OF INNATE IMMUNITY- PHAGOCYTES

1.2. Polymorphonuclear leukocytes
Basophil: Functions similar to eosinophils and mast cells i.e play a role in
both parasitic infections and allergies, segmented nucleus, granules
stained with basic dye such as hematoxylin (dark blue), less than 1%
How? FcR; granules: histamine, proteoglycan (heparin, chondroitin),
proteolytic enzymes (elastase, lysophospholipase)
Mast cells?

NTTH-HCMIU-IM-2014

NTTH-HCMIU-IM-2014

Exercise-1
WHO is WHO?
The five main WBC types

NTTH-HCMIU-IM-2014

CELLS OF INNATE IMMUNITYNK, mast cells and platelets

1. Phagocytic cells (Phagocytes):
1.1 Mononuclear phagocytes: Monocytes (circulating cells) and Macrophages
(reside in tissues)
1.2 Polymorphonuclear granulocytes (Polymorphs): with lobed, irregularly
shaped- polymorphic- nucleus
2. NK cells, mast cells and platelets

NTTH-HCMIU-IM-2014

Mast cells
Not found in circulation (i.e. tissue only, located in mucous membrane and connective
tissue throughout body), very similar to basophils in their characteristics; Major effector cell
in allergy; Modulation of initial immune response

NOW is yes

NTTH-HCMIU-IM-2014

Platelets
= cell fragments derived from megakaryocytes in the BM; 10A11 platelets/ day, 30%
stored in spleen
Involves in blood clot- platelet plug and fibrin clot formation (granules- alpha and delta,
microtubules, actin/ myosin filaments),
Alpha granules: insulin-like growth factor 1, platelet-derived growth factor, TGFp, platelet
factor 4 (which is a heparin-binding chemokine) and other clotting proteins (such as
thrombospondin, fibronectin, and von Willebrand factor).
Dense granules (also known as dense bodies or delta granules) are specialized secretory
organelles: ADP, ATP, ionized calcium (which is necessary for several steps of the
coagulation cascade), histamine and serotonin.

NTTH-HCMIU-IM-2014

WHAT PLATELETS DO....

1. Stick to damaged blood vessels
- requires von Willebrand factor
2. Spread out to cover damaged area
3. Activate and release contents
- partly blocked by Aspirin
4.Aggregate:
constriction

Cause
Cause

blood

vessel

retraction

of

clot to draw wound edges together

NTTH-HCMIU-IM-2014

NK cells (natural killer)

NK- Large granular lymphocytes (LGLs), ~ 15% lymphocytes, do not
express BCR and TCR. Able to recognize and kill via apoptosis virusinfected cells and certain tumor cells involving activating and inhibitory
receptors of NK cells (interact with MHC-I) and ALSO able to kill via ADCC
mechanism.

1: The small lymphocyte (B, T) has no granules, round nucleus and high N:C ratio
2: The LGL has azurophilic granules, indented nucleus and lower N:C ratio; CD16+
CD56+ (NK cells), others (~ 5% Th, 30-50% Tc)
NTTH-HCMIU-IM-2014

Myeloid
lineage

INNATE

Lymphoid
lineage

ADAPTIVE

Derived from common progenitor cell in bone marrow

- Pluri-potent hematopoietic stem cell
NTTH-HCMIU-IM-2014

CELLS OF THE ADAPTIVE IMMUNITY

Lymphocytes: 6-10 um in diameter, 20- 50% of circulating leucocytes
T cells (CD3, CD4/ CD8) and B cells (CD19)
Marker molecules: communicating with environment

NTTH-HCMIU-IM-2014

THE CLUSTER OF DIFFERENTIATION (CD)

A protocol for identification and investigation of cell surface
molecules
CD number assigned on basis of 1 cell surface molecule
recognized by 2 specific monoclonal antibodies
CD nomenclature established in 1982
1st International Workshop and Conference on Human
Leukocyte Differentiation Antigens (HLDA)

NTTH-HCMIU-IM-2014

THE CLUSTER OF DIFFERENTIATION (CD)

Examples of CD markers on leukocytes
Granulocyte

CD45+, CD15+

Monocyte

CD45+, CD14+

T lymphocyte

CD45+, CD3+

T helper lymphocyte

CD45+, CD3+, CD4+

T cytotoxic lymphocyte

CD45+, CD3+, CD8+

B lymphocyte

CD45+, CD19+

Natural killer cell

CD45+, CD16+, CD56+, CD3-

NTTH-HCMIU-IM-2014

CELLS OF THE ADAPTIVE IMMUNITY

T cells:
85-95 % of circulating lymphocytes T cell subsets: based on TCR and CD
Th (CD4+): - MHC-II
Th1 secretes IL-2;
Th2 secretes IL-4, IL-5, IL-6, IL-10
Tc (CD8+): -MHC-I
NKT cells: have both T cell markers and
some NK cell markers

NTTH-HCMIU-IM-2014

CELLS OF THE ADAPTIVE IMMUNITY

B cells:
5-15 % of circulating lymphocytes;
BCR complex: Immunoglobulin (Ig)+ CD79a/b (Iga/b)
Most express sIgM and sIgD
Some of circulating B cells (~ 10%) express IgG, IgA or
IgE while many more of localizing B cells express these markers
B-1 cells: respond well to TI Ag (Ag stimulate B cells without T cell help),
secrete natural Abs- of low avidity, but unusually they are poly reactive and
are found at high concentration in the adult serum, mostly IgM but also
some IgG and IgA
B-2 cells (conventional B cells): respond to TD Ag

NTTH-HCMIU-IM-2014

CELLS OF THE ADAPTIVE IMMUNITY

B cells: differentiate into Ab-secreting plasma cells:
B cells^ Ab-forming cells (AFCs)^ Plasma cells (infrequent in circulation,
restricted in secondary lymphoid organs and BM)
Plasma cells: survive for only a few days

NTTH-HCMIU-IM-2014

Myeloid
lineage

INNATE

Lymphoid
lineage

ADAPTIVE

Derived from common progenitor cell in bone marrow

- Pluri-potent hematopoietic stem cell
NTTH-HCMIU-IM-2014

Antigen presenting cells

APCs: Antigen presenting cells- a variety of cell types that
carry antigens in a form that can stimulate lymphocytes

NTTH-HCMIU-IM-2014

Who are APCs?

1. Dendritic cells: Cells with dendriform (star shaped) morphology
- DC (iDC) (interdigitating) found primarily in skin, lymph nodes and
spleen and within or underneath most mucosal epithelia and also in
thymus.
from BM, interact mainly with T cells
DC1- myeloid DC- from myeloid progenitor,
DC2- plasmacytoid DC- from lymphoid progenitor)
- FDC (follicular): found in the primary and secondary follicles of B cell
area in lymph nodes, spleen and MALT not from BM, interact with B
cells. In the B cell follicles of lymphoid organs and tissues, FDCs bind
small immune complex (IC, iccosomes) and present to specific B cells.

NTTH-HCMIU-IM-2014

Who are APCs?

DCs

Different kinds of antigen-presenting cells (APCs)

NTTH-HCMIU-IM-2014

Dendritic cells- DC

NTTH-HCMIU-IM-2014

Who are APCs?

2. Macrophages
3. B cells
Professional antigen- presenting cells: those that express MHC class II
(Major Histocompatibility Complex class II ) molecules

NTTH-HCMIU-IM-2014

Non- professional APCs

A non-professional APC does not constitutively express
MHC class II proteins required for interaction with naive T cells; these are
expressed only upon stimulation of the non- professional APC by certain
cytokines such as IFN-Y.
Non-professional APCs include:
- Fibroblasts (skin)
- Thymic epithelial cells
- Thyroid epithelial cells
- Glial cells (brain)
- Pancreatic beta cells
- Vascular endothelial cells

NTTH-HCMIU-IM-2014

Cells of IS- Requirement

Origin
Characteristics
Functions/ Roles and position in innate/adaptive immune
response

NTTH-HCMIU-IM-2014

Looking back to the previous lecture

COMPLETE BLOOD COUNT WITH DIFFERENTIAL (CBC WITH DIFF)
References Ranges
Erythrocytes (RBC)

4.0 to 5.4*

M/uL

Thrombocytes (Platelets)

145 to 400

K/uL

Leukocytes (WBC)
Neutrophils
Band neutrophils

4.8 to 10.8
40 to 74
0 to 9

K/uL
%
%

Eosinophils

0 to 6

Basophils

0 to 1

Lymphocytes
Monocytes

15 to 47
0 to 12

%
%

*: You have the chance to check this number of your own blood in the practical course by simply counting

NTTH-HCMIU-IM-2014

SUMMARY

NTTH-HCMIU-IM-2014

ORIGIN OF CELLS OF THE IMMUNE SYSTEM

Derived from common progenitor cell in bone marrow
Pluripotent hematopoietic stem cell

Progenitor Stem Cells

1. Erythroid lineage
Erythrocytes and Megakaryocytes

2. Myeloid lineage
Monocyte/macrophage, dendritic cells, PMNs, mast cells

3. Lymphoid lineage
Small and large lymphocytes
NTTH-HCMIU-IM-2014

CELLS OF INNATE AND ADAPTIVE IMMUNITY

2.1 Myeloid Lineage- I
Neutrophil
Principal phagocytic cell of innate immunity
Eosinophil
Principal defender against parasites
Basophil
Functions similar to eosinophils and mast cells
Referred to as
Polymorphonuclear leukocytes (PMNs)
Nuclei are multilobed (2 to 5)
Granulocytes
Cytoplasmic granules
NTTH-HCMIU-IM-2014

CELLS OF INNATE AND ADAPTIVE IMMUNITY

2.2 Myeloid lineage-II
Monocytes
Leukocytes with bean shaped or brain-like convoluted nuclei
Circulate in blood with half life of 8 hours
Precursors of tissue macrophages

Macrophages
Mononuclear phagocytic cells in tissue
Derive from blood monocytes
Participate in innate and adaptive immunity

NTTH-HCMIU-IM-2014

CELLS OF INNATE AND ADAPTIVE IMMUNITY

2.3. Myeloid lineage-III
Dendritic cells
Cells with dendriform (star shaped) morphology
Interdigitating reticular cells (synonym)
Capture and present antigens to T lymphocytes
Mast cells
Located in mucous membrane and connective tissue
throughout body
Major effector cell in allergy
Modulation of initial immune response
NTTH-HCMIU-IM-2014

COMPLETE BLOOD COUNT WITH DIFFERENTIAL

(CBC WITH DIFF)
References Ranges
Erythrocytes (RBC)

4.0 to 5.4*

M/uL

Thrombocytes (Platelets)

145 to 400

K/uL

Leukocytes (WBC)

4.8 to 10.8

K/uL

Neutrophils
Band neutrophils

40 to 74
0 to 9

%
%

Eosinophils

0 to 6

Basophils

0 to 1

15 to 47

0 to 12

Lymphocytes
Monocytes

*: You have the chance to check this number of your own blood in the practical
course by simply counting
NTTH-HCMIU-IM-2014

http://upload.wikimedia.org/wikipedia/commons/b/b5/Reference_ranges_for_blood_tests_-_white_blood_cells.png

Refer to CASE STUDY!

NTTH-HCMIU-IM-2014

CELLS OF INNATE AND ADAPTIVE IMMUNITY

3. Lymphoid Lineage
3.1. Large lymphocytes (large granular lymphocytes)
Natural killer (NK) cells (CD16, CD56)
Innate immunity to viruses and other intracellular pathogens
Participate in antibody-dependent cell-mediated cytotoxicity (ADCC)

3.2. Small lymphocytes

B cells (CD19)
T cells (CD3, CD4 or CD8)
Adaptive immunity

Lymphocytes refers to small lymphocytes

NTTH-HCMIU-IM-2014

1. Cell of the IS
Remember this IMAGE? Lets revise together

NTTH-HCMIU-IM-2014

Cells of IS- another illustration

Fetal liver/ BM

HSC/ CFU-GEMM

CFU-GM
NKT

Color Atlas of Immunology- 2003

NTTH-HCMIU-IM-2014

Discussion Topics
1. Explain more clearly the question at P30-DIR,
2. Methods to identify cell populations
3. Methods to isolate cell populations
4. How T cell can distinguish between self and nonself Ags?
5. Homework: critical thinking 1 & 2

NTTH-HCMIU-IM-2014

Homework check
Q&A
Any Question from you?

NTTH-HCMIU-IM-2014

Discussion Topics
1. Why do warm-blooded, long-lived animals require particularly
complex immune defense?- p4-DIR
2. Why would removal of Ag lead to the decline in an immune
response?- p14-DIR
3. Lets go to Critical Thinking, p18-DIR. Rewrite our discussion
as a homework.
And many more to explore in the DIR textbook

NTTH-HCMIU-IM-2014