Haematopoiesis

How does the immune system do the job?

A Network of Specialized Cellular Components & Organs
Hematopoietic Stem Cells

Major Cell Types

 Cells & Organs of Immune system
• Many cells, organs and tissues of immune system
are found throughout the body.
• Primary lymphoid organs: provide appropriate
microenvironment for development and maturation
of lymphocytes. (Thymus, Bone marrow)
• Secondary lymphoid organs trap antigen from
nearby tissues and are sites where mature
lymphocytes can interact effectively with antigen.
(Lymph node, Spleen, MALT)
 Haematopoiesis
• Cells of immune system present in blood and
lymph and in various lymphoid organs.
• Broadly referred as leucocytes (WBCs)
• Process of formation and maturation of blood
cells: Haematopoiesis.
• Sites of haematopoiesis: embryonic yolk sac during
first weeks of development
• 3rd -7th month of gestation: HSCs migrate to fetal
liver and spleen
• After that differentiation of HSCs occurs in bone
marrow.
 HSCs
• Studying HSCs is difficult
 A very scarce population
 In vitro culture is difficult
Because of self renewal capacity HSCs are
maintained at stable level throughout life.
Can display enormous proliferative capacity
whenever there is a demand for haematopoiesis.
(as demonstrated by lethal X-ray irradiation of
mouse)
The Journal of Clinical Investigation     
http://www.jci.org      Volume 116      Number 5     
May 2006
 Dendritic
cell

Dendritic
cell
 Stem Cell Definition
• A cell that is capable of self-renewing,
and converting into different cell type
 Stem Cell Definition
• Self-renewal – the ability to form new stem
cells
• Differentiation – the ability to give rise to a
heterogeneous progeny of cells
• Homeostatic control – the ability to modulate
and balance differentiation and self-renewal
Embryonic Stem Cells vs Adult Stem Cells

Fertilization Embryonic
Stem Cells
Pluripotent
Stem cells

Adult Stem Cells

Hematopoietic Nerve Liver

An Example - Hematopoietic Stem Cells (self-renewal, and
differentiation)

Tan et al., Laboratory Investigation 2006,86,1203-1207

Haematopoietic stem cell (HSC)
 Differentiation of HSCs
• Early in haematopoiesis a HSC will form either
a lymphoid progenitor or a myeloid
progenitor.
• Progenitor cells have lost the capacity to self
renew and they are now committed to
develop into one particular lineage.
• CMP and CLP
Haematopoietic inducing microenvironment
(HIM)

Stromal cells: fat cells,

fibroblasts,
macrophages,
endothelial cells,

Cellular matrix and

growth factors that
promote growth and
differentiation
• During haematopoiesis erythrocytes and many
different kinds of WBCs descend from few
HSCs.
• A complicated process which involves
hierarchy of many precursor populations.
• Why such a complex process?
 Two major influences shape stem cell
differentiation:

1. Expression of specific transcription factors for

lineage commitment

2. the CYTOKINES AND GROWTH FACTORS they are

exposed to
Lineage commitment & proliferation
 Haematopoietic homeostasis
• Haematopoiesis is a complex process where
mature blood cells are produced at the same
rate as they are lost.
• Every blood cell has a life span.
• To maintain steady state level an average
human being must produce 3.7X1011 WBCs
per day.
 Enrichment of HSCs and clinical applications

• HSCs can be identified by specific

markers/antigens.
• CD34 a marker found on 1% of haematopoietic
cells that includes stem cells.
• HSCs can be purified based on this principle and
used to reconstitute a patient’s entire
haematopoietic system.
• Clinical applications: HSC transplantation for
immunodeficiency or defective haematopoietic
system.
Haematopoietic tree
 Blood components
• Primary components of blood:
• Plasma
• Cells
• Erythrocytes (45-50% of total blood volume)
• Leucocytes (1-2% of total blood volume)
• Specific gravity of blood: 1.05-1.064
• Serum obtained after coagulation of blood
 Blood fractionation
• Blood fractionation is the process of 
fractionating whole blood, or separating it
into its component parts. This is typically
done by centrifuging the blood.
• The resulting components are:
Erythrocytes
 Leucocytes (Granulocytes, Lymphocytes,
Monocytes)
• Granulocytes (Neutrophils, basophils, Eosinophils)
on basis of cellular morphology and cytoplasmic
staining characteristics.
• Neutrophil: multilobed nucleus (PMN), granulated
cytoplasm, stains with both acid and basic dyes
• Eosinophil: bilobed nucleus, stains with acid dye
eosin
• Basophil: lobed nucleus, stains with basic dye
methylene blue, heavily granulated cytoplasm
Neutrophils (PMNs)
Make up 50-70% of all leucocytes.
Circulate in blood for 6-8 hrs
before they migrate into tissues
where they have a life span of
few days (1-4 days)
Eosinophils & Basophils
Eosinophils: phagocytic ,
defence gainst parasitic
organisms
Basophils: non phagocytic,
involved in allergic reactions

Mast cells: present in variety of

tissues, skin, various organs,
mucosa
Dendritic cells

Plasmacytoid DCs (pDCs)

 Dendritic cells
• Versatile cells
• Perfoem antigen capture in one location and
antigen presentation in another.
• Outside lymph nodes they exist in immature
form act as “sentinels of immune system”
• After antigen capture undergo a maturation
programme and become Mature DCs can
present antigen to T cells.
 Mononuclear phagocytes (Monocytes)
• Mononuclear phagocytic system: monocytes
in blood and macrophages in tissues.
• Monocytes and lymphocytes are together
referred as mononuclear cells (PBMCs).
• Tissue resident macrophages:
Monocytes and Macrophages
 Lymphoid cells
• Constitute 20-40% of WBCs and 90% of cells in
lymph.
• Approximately 1011-12 lymphocytes in human
body.
• Circulate continuously in blood and lymph
• Can migrate into tissue spaces and lymphoid
organs
• Three major populations: T, B, NK
Fate of antigen activated lymphocytes (naïve,
effector , memory)
 T cells & B cells
• T cells: Th cells (CD4) , Tc or CTLs (CD8) and Treg
(CD4+, CD25+) cells
• Th:Tc ratio 2:1
• B cells: naïve B cells, plasma cells which secrete
antibodies
• Difference in which B and T cells recognize antigen
• T cells are MHC restricted, recognize antigen only
when it is bound to another protein called MHC
 NK cells (CD16)
• Large granular lymphocytes
• Cytotoxic against tumour cells and virus
infected cells.
Identify cells