Histology Department

Prepared by: LokDonLub

1
 INTRODUCTION
• The immune system: a system of cells that able
to distinguish "self" (organism's own molecules)
from "non-self" (foreign substances)1

• The cells of the immune system :1

1. Distributed throughout the body (in blood, lymph,
epithelial & conn. tissues)
2. Arranged in small spherical nodules → lymphoid
nodules (in conn. tissues & inside several organs)
3. Organized in larger lymphoid organs (lymph nodes,
spleen, thymus & bone marrow)
Cells of the Immune System
 Cells of the Immune System
• The primary cells in the immune response:1
1. Lymphocytes
2. Plasma cells
3. Mast cells
4. Neutrophils
5. Eosinophils
6. Cells of the mononuclear phagocyte system

• Antigen-presenting cells: a group of diverse cell

types, assist other cells in the immune response
– Lymphocytes
– Macrophages
– Dendritic cells
 1
LYMPHOCYTE 2
NEUTROPHIL 3
EOSINOPHIL

4
MONOCYTE 5 CELL
PLASMA 6
MACROPHAGE
 Lymphocytes
• Classified as B, T, or natural killer (NK) cells

• Key features of B & T lymphocytes → The receptor on cell’s

surface

• The B & T cells:

– Able to recognize a specific epitope
– Differ based on life history, surface receptors, & behavior during an
immune response
– Morphologically indistinguishable by light or electron microscope,
but have different surface proteins (markers) → distinguished by
immunocytochemical methods

• B & T cells continuously move from one location to another

(lymphocyte recirculation)
EXAMPLE OF IMMUNOHISTOCHEMISTRY MARKER
TO IDENTIFY B OR T CELL 5
A non-glycosylated phosphoprotein expressed on the
M CD20
C membrane of B-cells
A
E A heterodimeric glycoprotein signal transduction
R molecule that associates with membrane
B L
K immunoglobulin → provide an almost perfect B-cell
L CD79a
E lineage marker because expressed throughout B-cell
S differentiation
R

An excellent marker of T-cells & expressed

M CD5
in most T-cell lymphomas
C
A A marker of helper/inducer T-cells & their neoplasms,
E
R CD4 also present on histiocytes
T L
K
L A marker of suppressor/cytotoxic T-cells & neoplasms
E
S CD8 derived from them
R
 B Lymphocytes
• The surface receptors able to recognize antigens are
monomeric molecules of IgM → Each B cell is covered by
about 150,000 molecules of IgM

• The B lymphocyte contact with the epitope (activated B

cells) change into:
– Plasma cells (after several cycles of cell proliferation, followed
by a redifferentiation) → secretes antibodies against the same
epitope
– Long-lived B memory cells → able to react very rapidly to a
second exposure to the same epitope

• Activation of B cells requires assistance of T helper cells

 T Lymphocytes
• 65–75% of blood lymphocytes

• All T cells have a surface molecule (T cell receptor-

TCR) → recognize only epitopes (mostly small
peptides) that form complexes with special proteins
on cell surface of other cells (proteins of the major
histocompatibility complex)

• Three important subpopulations of T cells:

– Helper T cells (CD4+ T cells)
– Cytotoxic T cells (CD8+ T Cells)
– Regulatory T cells (CD4+CD25+ T Cells)
 T Lymphocytes cont’d
• Helper T cells → Produce cytokines:
– Promote differentiation of B cells into plasma cells
– Activate macrophages to become phagocytic
– Activate cytotoxic T lymphocytes
– Induce many parts of an inflammatory reaction

• Cytotoxic T cells → Act directly against foreign cells

or virus-infected cells by two main mechanisms:
– Attach to cells to be killed & release perforins that create
holes in cell membrane → cell lysis
– Attach to a cell & kill it by programmed cell death, or
apoptosis
 T Lymphocytes cont’d
• Regulatory T cells:
– Crucial roles in allowing immune tolerance,
maintaining unresponsiveness to self-antigens &
suppressing excessive immune responses
– Produce peripheral tolerance, which backs up the
central tolerance emerging in the thymus
 Natural Killer Cells
• Lack the marker molecules characteristic of B
& T cells

• About 10–15% of the lymphocytes of

circulating blood

• Attack virus-infected cells, transplanted cells

& cancer cells without previous stimulation →
innate immune response
 Antigen-Presenting Cells (APCs)
• Found in many tissues & constitute a heterogeneous population
of cells:
– Dendritic cells
– Macrophages
– B lymphocytes

• A common feature of APC: class II MHC molecules on their

surface

• Dendritic cells found in lymphoid organs, abundant in epidermis

& many mucosa (Langerhans cells)

• The Langerhans cells of the epidermis:

– For trapping antigens that enter the epidermis
– Have many processes & upon capturing antigens, they retract the
processes, move toward the dermis, & enter a lymphatic vessel
Lymphoid Tissue & Organs
LYMPHOID ORGANS : 1, 3
• Primary/central organs :
Thymus & Bone Marrow 
responsible for
development & maturation
of lymphocytes

• Secondary/Peripheral
organs: Lymph nodes,
Spleen, Tonsils, solitary
nodules, Peyer’s Patches of
ileum, Appendix
Approximate Percentage of Lymphocytes
in Lymphoid Organs1
T lymphocytes B Lymphocytes
Lymphoid organs
% %
Thymus 100 0
Bone marrow 10 90
Spleen 45 55
Lymph nodes 60 40
Blood 70 30
 Lymphoid Tissue
• A connective tissue characterized by a rich supply
of lymphocyte:
– Free within the regular conn. Tissue
– Surrounded by capsules, forming the lymphoid organs

• Basically made up of:

– Free cells &
– Rich network of reticular fibers of type III collagen
supporting the cells

• The reticular fibers produced by a fibroblastic cell

→ RETICULAR CELL
 Lymphoid Tissue cont’d
• Lymphoid nodules or lymphoid follicles: 1
– Groups of lymphocytes (primarily B lymphocytes) arranged as
spherical masses found in nodular lymphoid tissue
– Vary widely in size (few hundred µm to 1 mm)
– Free in many conn. tissues & within lymph nodes, spleen, &
tonsils
– NOT FOUND in thymus which contains only T cells

• Lymphoid nodules that found in mucosa of:1

– The digestive system (tonsils, Peyer's patches, & appendix)
– The respiratory system
– The reproductive system
– The urinary system
→ Collectively known as MUCOSA-ASSOCIATED LYMPHOID
TISSUE (MALT) & considered a lymphoid organ
 Germinal Center
• An area in central portion of activated lymphoid nodules
due to activation & proliferation of B lymphocytes by
antigen → stains lighter

• After completion of initial immune response → germinal

center may disappear

• Contain follicular dendritic cell (FDC, distinct from the

dendritic APCs), with many extremely fine processes in
the surfaces

• Antigens bind surface proteins of FDCs is retained for

extended periods (months to years) for interaction with B
lymphocytes
 Germinal Center

Activated Lymphoid Nodules

 A reactive
germinal follicle
showing:
the germinal
centre cells (left),
mantle cells
(centre)
& marginal zone
cells with
increased clear
cytoplasm (right).

Reactive germinal
follicle stained for CD21
to show follicular
dendritic cells
Lymphoid Organs: 1. THYMUS
 THYMUS
• A bilateral organ located in
mediastinum

• Has a conn. tissue capsule →

penetrates parenchyma &
divides into incomplete lobules
(continuity between cortex &
medulla)

• No afferent lymphatic vessels

& not a lymph filter

• Each lobule consist of:

– The cortex: a peripheral darkly
stained zone → richer in
lymphocytes than medulla
– The medulla: a central light zone
 Thymus: The Cortex
• The cortex composed of:
1. T lymphoblasts
(thymocytes): 97-98% of
developing T cells die by
apoptosis in cortex 
phagocytosed by
macrophages
2. Macrophages
3. Epithelial reticular cells
 Thymus: The Epithelial Reticular Cells
• Diverse morphologically→
squamous or stellate with long
processes

• Large euchromatic nuclei

• Intermediate keratin filaments

(tonofilaments) (+) → epithelial
origin

• Joined by desmosomes forming

cytoreticulum
• Occluding junctions of epithelial
reticular cells help to separate
between cortex & medulla
The cortical zone packed with
lymphoblasts, supported on a
meshwork of epithelial reticular cells
(arrowheads). X400. PT
3 Types of Epithelial Reticular Cells in Cortex 3

• Type I cells
• Type II cells
• Type III cells

• Functions of the cells:

– Completely isolate thymic
cortex & prevent developing
T Cells from contacting
foreign antibodies
– Presenting self antigens The relationship between epithelial reticular
(MHC I & II molecules) to cells and thymus lymphocytes. Note the
desmosomes and the long processes of
developing T Cells epithelial reticular cells extending among the
lymphocytes
Thymus: The Medulla
• Stain lighter than cortex 
less T cells population &
large number of epithelial
reticular cells

• 3 types of epithelial
reticular cells in medulla:
• Type IV cells
• Type V cells
• Type VI cells  Hassl’s
Body/Thymic Corpuscle
(found only in medulla,
cornified, even calcified,
unknown function)
 Thymus Vascular Supply1, 3
• The Blood-Thymus Barrier in the cortex created
by:→ prevent circulating antigens entering cortex
– Arterioles & continous capillaries with thick basal
lamina
– Flattened epithelial reticular cells type I with tight
junctions that sheathed arteriole & capillaries

• Self macromolecules crossed barrier  to select &

eliminate T cells react with self antigens  clonal selection
& clonal deletion

• No barrier in medulla

• T cells leave medulla via veins drainning the thymus

 Hormones In Thymus 1, 3
• Epithelial reticular cells produce :
– Thymosin
– Thymopoietin
– Thymulin
– Thymic humoral factor
 Facilitate T cell proliferation & expression of surface markers

• Other hormones influence T cells maturation :

– Corticosteroids  decrease T cells number in cortex
– Thyroxin  stimulates epithelial reticular cells to increase
thymulin production
– Somatotropin  promotes T cells development in thymus
cortex
 THYMUS INVOLUTION4
• Start after puberty

• Parenchym replaced
adipose tissue & conn.
tissue

• Decrease weights : 40 g
at puberty, 10-15 g late
in life

• After involution,
thymus still has its Elderly thymus
showing
function as a A severe atrophy
maturation place for T of parenchyma &
cells massive adipose
infiltration.
Lymphoid Organs: 2. LYMPH NODES
 LYMPH NODE
• Kidney shape, encapsulated
(Conn. Tissue Trabeculae), 2–10
mm in diameter

• A series of in-line filters →

Important in body's defense
against microorganisms & the
spread of tumor cells

• Have Afferent lymph vessel &

Efferent lymph vessel 1

• Hilum : concave depresion which

arteries & nerve enter, veins &
lymphatic vessels leave1,2
 LYMPH NODE
• Parenchym composed of T
cells, B cells, APCs,
macrophages,3 plasma
cells & reticular cells,
follicular dendritic cells1

• The cells arranged into: 1

– A Cortex
– A Medulla
– An intervening Paracortex

The cortex (C), The paracortex (P), & the medulla (M). Conn. tissue of the capsule (CT)
Trabeculae (T). Lymphoid nodules (LN) normally restricted to the cortex & the medulla
characterized by sinuses (MS) & cords (MC) of lymphoid tissue. X40. H&E.
 THE CORTEX
Consists of:
1. Many reticular cells,
macrophages, APCs, &
lymphocytes
2. Lymphoid nodules →
formed mainly of B
lymphocytes The capsule (C), subcapsular sinuses (S), lymphoid
nodules (N).
3. The subcapsular
sinuses → areas
beneath the capsule
4. The Cortical sinuses,
running between
lymphoid nodules, arise
from & share the
structural features of
the subcapsular sinuses
 THE PARACORTEX
• No precise boundaries

• Distinguished from outer

cortex by lack of B cell
lymphoid nodules &
accumulation of T cells
(determined by IHC)

• Venules in paracortex →
an important entry point
for lymphocytes moving
from blood into lymph
nodes
 THE MEDULLA
Two major components:
1. Medullary cords → branched cordlike extensions of lymphoid
tissue arising from paracortex, primarily B lymphocytes, plasma
cells & macrophages
2. Medullary sinuses → dilated spaces, contain lymph, lymphocytes,
often many macrophages, & sometimes granulocytes
 THE LYMPHOCYTE CIRCULATION
• Naive lymphocyte spend < ½ hour
in circulation before homing to
another lymphoid organ

• 2 main ports of entry into Lymph

Node :
1. By High Endothelial Venule
(HEV)
• Specialized venule, lined by cuboid
or high endothelial cells
• Found only in secondary lymphoid
organs except spleen
• Main site of B & T lymphocytes
(90%) entry from blood
2. By afferent lymph vessel
Photomicrograph of a high
endothelial venule in a lymph node.
Arrowheads indicate high
endothelial cells. The venule is
crossed by lymphocytes (arrows).
PT stain. High magnification.
 NORMAL
LYMPH NODE

SPORADIC BURKITT
LYMPHOMA:
A non-Hodgkin
Lymphoma, mainly in children &
young adult (30-50% of all
childhood lymphoma)
Lymphoid Organs: 3. SPLEEN
 SPLEEN
• Largest
3
lymphoid organ in body

• The normal weighs between

100 - 170 g & not palpable on
clinical examination

• Functions:
– Filtration of blood (the only
one) → defense against blood-
borne antigens
– Main site of destruction of aged
erythrocytes
– Production site of antibodies &
activated lymphocytes

• Surrounded by capsule (conn.

Tissue)→ trabeculae,
subdivide parenchyma or
splenic pulp
 SPLENIC PULP
• Composed of reticular tissue
containing reticular cells,
many lymphocytes, other
blood cells, macrophages &
APCs

• Two components of splenic

pulp:
– White pulp →consist of
lymphoid nodules &
periarteriolar lymphoid sheaths
(PALS)
– Red pulp → consists of blood-
filled sinusoids & splenic cords
(of Bilroth)
Marginal zone 3
• Separate white pulp to
red pulp
• Composed of Marginal
sinuses, plasma cells, T
cells, B cells,
macrophages, APCs
• Contain an abundance
of blood antigens 
plays major role in
immunologic activities
of spleen
 Blood Circulation of Spleen

Splenic artery → trabecular arteries → central arterioles enveloped by the periarteriolar

lymphoid sheath/PALS. → branches as penicillar arterioles → 2 circulation: a closed
circulation passing directly into splenic sinuses (S) or an open circulation, being
dumped from the vasculature into the lymphoid tissue of the red pulp's splenic cords
Red Pulp :1, 3 Consist of:
• Splenic Cords /
Billroth’s Cords  The red pulp composed
Reticular cells, of splenic venous
macrophages, T cells, sinusoids (S) & splenic
B cells, plasma cells, cords (C),

blood cells
• Splenic Sinusoids
with unusual
elongated endothelial
cells (Stave Cells) →
w/ special properties
that allow selection
of healthy red blood
cells
The venous sinuses in the
red pulp (S) lined by
endothelial cells (arrows)
with large nuclei bulging
into the sinusoidal
lumens (the stave cells)
→ have special properties
that allow selection of
healthy red blood cells in
the splenic cords (C).
X100. H&E.
SPLENOMEGALY

Principal
Causes
Thalasemia: Hypochromic, microcystic
eritrocytes, poikilocytosis, target cells
 Lymphoid Organs:
4. Mucosa-Associated Lymphoid Tissue:
A. Tonsils
B. Gut-Associated Lymphoid Tissue
C. Bronchus-Associated Lymphoid Tissue
Collectively is one of the largest lymphoid organs,
containing up to 70% of all the body's immune cells
 MALT: TONSILS
• Partially
encapsulated
lymphoid tissue1

• Lying beneath & in

contact with
epithelium of the
oral cavity & pharynx

• Based on location :
palatine,
pharyngeal, lingual
tonsils1
 MALT: PALATINE TONSILS
• A pair, in posterior soft
palate1

• Consist of : 1, 2, 3
– Stratified squamous
Epithelium
– A band of lymphoid nodule
with germinal center
– Crypts :
• Invagination of epithelium
• 10-20 crypts/tonsil
• Contain food debris, dead
leucocytes, desquamated of
epithelial cells,bacteria etc
– Capsule  partially at the
base
• Single in posterior MALT: PHARYNGEAL
nasopharynx1, 2
TONSILS
• Consist of :1, 2, 3
– Pseudostratified
ciliated columar
epithelium
– Lymphoid nodules
– No crypts, only
shallow
longitudinal
infolding called
pleats
– Thinner capsule
than T. Palatina
• Smaller & more
numerous than MALT: LINGUAL TONSILS
other tonsils

• At base of tongue

• Consist of :1, 2, 3
– Stratified
Squamous
Epithelium
– Lymphoid nodules
– Each lingual
tonsils has a
single crypts
MALT: GUT-ASSOCIATED LYMPHOID TISSUE 1, 3
• Non capsulated
lymhoid nodule

• extends along the

entire gastrointestinal
tract,

• Peyer patches: large

aggregates of
lymphoid follicles in
the ileum, each
consisting of 10–200
nodules & bulging
into the gut lumen
with no connective
tissue capsule
The simple follicle-associated epithelium (FAE) covering nodules
contains M cells (characterized by a large basal intraepithelial pocket
open to the underlying lymphoid tissue through a porous basement
membrane) → continuously sample antigens & microorganisms in the
intestinal lumen
Often MALT consists of small
accumulations of lymphoid
cells or one to a few lymph
follicles beneath the
epithelium & possibly
extending into the submucosa
MALT: BRONCHUS-ASSOCIATED LYMPHOID TISSUE 1, 3
• Similar to peyer’s
patch except locate at
walls of bronchi &
bronchioles

• Aggregates of
lymphoid follicles

• Epithelial covering A collection of lymphocytes in the

change from connective tissue of the bronchiolar
pseudostratified mucosa, an example of mucosa-associated
columnar ciliated lymphoid tissue (MALT). Pararosaniline—
toluidine blue (PT) stain. Low
epithelium to M Cells magnification
 REFERENCES :
1. Junqueira's Basic Histology, Twelfth Edition. Anthony L.
Mescher, PhD. The McGraw-Hill Companies, 2010.
2. Essentials of Human Histology, 2nd Edition, William J. Krausse
PhD, Little Brown & Company (Inc), 1996. Pp 197-228
3. Color Textbook of Histologi, 2nd edition, Gartner LP, Hiatt JL,
WB Saunders Company, Philadelphia, Pennsylvania, 2001. Pp
273-299
4. Consise Histology, 2nd edition, Don W Fawcett, Ronald P
Jensh, Arnold publisher, London, 2002. Pp 148-161
5. Diagnostic Lymph Node Pathology. Dennis H. Wright. Bruce J.
Addis, Anthony S.-Y. Leong. Hodder Arnold, 2006.
6. Rubin’s Pathology. 6th ed. Rubin R, Strayer DS. Lippincott
Williams & Wilkins, 2012
 2

3
1. Area pointed by arrow # 2 is...... MEDULLA
2. Arrow # 3 are..... LYMPHOID NODULE/GC
3. The Organ shown here is.......LYMPH NODE
1. The structure in the circle is.... WHITE PULP
2. The organ shown here is....... SPLEEN
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