HISTOLOGY

SHIFT 3
Lymphoid System and Immunity
Albert M. Luna, MD
06 FEB 2023
AY 2022-2023
14
TABLE OF CONTENTS • Situated in the left upper quadrant of the
abdomen
1. Immune System ……………………………………………1 Spleen • Location where T and B lymphocytes may
2. Protective Surface Mechanisms ………………………..1 interact with blood-borne antigen and
2.1. Natural Barriers undergo stimulation and cell division
3. Innate Immune System …………………………………...2 • Mucosa-associated lymphoid tissue
3.1. Components • Includes the tonsils and adenoids in the
4. Adaptive Immune System ………………………………..2 oropharynx, Peyer’s patches and lymphoid
5. Lymphocytes and Antigen-Presenting Cells …………3 aggregates of the small and large intestines,
MALT
5.1. Lymphocytes respectively, and a diffuse population of
5.2. Antigen-Presenting Cells lymphocytes and plasma cells in the
6. Immune Response ………………………………………...5 mucosae of the gastrointestinal,
6.1. Recognition of Antigen respiratory and genitourinary tracts
6.2. Activation of Immune System
6.3. Generation of Effector Mechanisms
6.4. Termination of Immune Response
7. Lymphoid Organs …………………………………………7
7.1. Bone Marrow 2. PROTECTIVE SURFACE MECHANISMS
7.2. Thymus • First line of defense
7.3. Lymph Nodes
7.4. Mucosa-Associated Lymphoid Tissue
• Provide Excellent Protection (while intact)
7.5. Spleen
8. Tumors of the Immune System ………………………..23
9. Slide Review ………………………………………………23 2.1. Natural Barriers
9.1. Thymus
9.2. Lymph Nodes Surface layer of keratin constitutes an
9.3. Spleen impenetrable barrier to most
Skin
microorganisms unless breached by
injury such as a cut, abrasion, or burning
LEGEND Mucous
Protected by various anti-bacterial
★ Take note / Important ☛ Prof verbatim substances:
Surfaces
• Defensins – short anti-microbial
✎ Textbook info ✂ Previous/Other trans info (conjunctiva peptides found in surface mucus
and oral
• Lysozymes – secreted in tears
1. IMMUNE SYSTEM cavity)
and saliva
• All living tissues are subject to the threat of Respiratory Surface mucus produced by goblet
pathogens that may invade the body, multiply and Tract cells, and removed by ciliary action
destroy functional tissues, causing illness and Stomach Highly acidic environment inhibits
potentially death and Vagina growth of pathogens in these sites
• Three main lines of defenses have evolved in
response to these threats:
o Protective Surface Mechanisms
o Innate Immunity
o Adaptive Immunity
✎ ORGANS OF THE IMMUNE SYSTEM (p. 198)
The cells of the adaptive immune system form specialized
lymphoid tissues and also constitute a significant component of
other tissues such as the gastrointestinal tract.
• Situated in the anterior mediastinum
Thymus • site of maturation of immature T
lymphocytes
Bone • Home of lymphocyte stem cells
Fig. 2.1.1. Entry of pathogens into skin and mucosae
Marrow • Site of B lymphocyte maturation
• Found at the junctions of major lymphatic
• When protective surface mechanism fails, the two
vessels other main types of defense mechanisms are
Lymph • Sites where both T and B lymphocytes may activated.
Nodes interact with antigen and APCs from the
circulating lymph leading to lymphocyte
activation and cell division

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3. INNATE IMMUNE SYSTEM 4. ADAPTIVE IMMUNE SYSTEM

• Provides rapid reaction
• Same magnitude of response each time
• No learning
• Causes a pathological condition known
inflammation
• Slower Initial primary response (3-5 days)
• Has the ability to learn
o Greater, more specific and faster response
as o Life long immunity (via past infection or
vaccination)

Acute Inflammation
• Characterized by vascular changes
o Dilatation
o Increased permeability
o Increased blood flow
o Formation of fibrin-rich inflammatory exudates

3.1. Components of Innate Immune System

COMPONENTS ACTIONS
• Phagocytosis of pathogenic
organisms
Neutrophil
• Secretion of cytokines,
Polymorphs Fig. 4.1. Slower response of adaptive immune system
Neutrophil Extracellular Traps
(NETs), and PRMs Adaptive Immunity Kills Pathogen In Two Ways
• Phagocytosis of pathogenic CELLULAR RESPONSE HUMORAL RESPONSE
organisms B cell (antibody
• T cell mediated
Removal of foreign material and mediated)
dead cells
Macrophages
• Secrete cytokines & interleukins
• Present antibodies to lymphocytes
• Not just a phagocyte but also an
antigen-presenting cell (APC)
• Destroy large multicellular
Eosinophils pathogens (i.e. intestinal worms)
and modulate allergic response
Natural Killer • Recognize and kill virus-infected
Cells and cancer cells These 2 mechanisms may combine in a concerted process
• Opsonizes organisms to facilitate
Adaptive Immunity Amplifies the Innate Immune
phagocytosis
Response
• Chemotaxis; Chemoattractant for
• Antibodies produced by B-cells may coat bacteria
Complement various cells
(opsonization), facilitating phagocytosis
• Cell lysis; MAC (membrane
• Antigen-antibody complexes can directly activate the
attack complex) kills cells by
puncturing plasma membrane complement cascade
• Plasma protein increased in
inflammation/infection promote
Acute-phase
defense against pathogens
Proteins
• Example: C-reactive Protein
(CRP) which acts as an opsonin
• Recruit cells and activate cells of
innate and adaptive immune
systems (chemoattractant)
Chemokines • Induce differentiation of cells to
more active and effective
subtypes
• Wound healing and angiogenesis
• Signaling molecules produced by
macrophages, dendritic cells, Fig. 4.1. Opsonization of antigens by antibodies to prime for
Interleukins phagocytosis
lymphocytes
• Regulate Immune system

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Adaptive Immune System Can Be Controlled By The

Innate Response
• T lymphocytes require the services of antigen-
presenting cells like macrophages for activation

Fig. 5.1.1. Round ovoid nucleus occupying 90% of cell volume

with a thin rim of basophilic bluish cytoplasm

Fig. 4.2. APC activating T-helper cell

5. LYMPHOCYTES AND ANTIGEN PRESENTING CELLS

5.1. Lymphocytes
• 20% to 50% of white blood cells in circulation
• Majority are small lymphocytes
o 6-9 μm (about the same size as erythrocyte)
• 3% are large lymphocytes (9-20 μm)
• Constantly patrol the body
o Circulate in blood, lymph and other ECF fluids
o Pauses occasionally in organized lymphoid
tissues
• Lymphocytes live in secondary lymphoid organs
o Strategically placed to optimize the chances of
meeting of an antigen meeting a potentially Fig. 5.1.2. Lymphocyte distribution in body
reactive lymphocyte
• Facilitate lymphocyte activation

5.2. Lymphocyte Receptors

T CELL RECEPTOR B CELL RECEPTOR

Fig. 5.2.1. Lymphocyte antigen receptors (TCR) Fig. 5.2.2. Lymphocyte antigen receptors (BCR)
B cell receptor is made up a “Y” shaped surface immunoglobulin
+ accessory molecules

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• Lymphocytes produce a huge range of antigen receptors makes the adaptive immune system highly effective in
recognizing a variety of pathogenic organisms in nature
5.1.1. T Lymphocytes

• Produced in the bone marrow

• Mature in the thymus
o Proliferation
o Rearrangement of TCR genes
o Acquisition of the surface receptors and accessory
molecules needed to become mature T cells
• Develop state of “self tolerance”
o Apoptosis of T cells that react to self-antigens
o Prevents autoimmunity
• T cells may develop into one of the functional subsets
depending on the mixture of cytokine sand interleukins
to which they are exposed Fig. 5.1.1.1. Subtypes of T helper cells

FUNCTIONAL SUBSETS OF T CELLS

SUBSET FUNCTION OTHER
• “Help” other cells (B cells, Cytotoxic T cells, • Further subdivision into:
Macrophages) perform their effector functions o TH1 (cell mediated reaction)
• Achieves this by secretion of mediators called o TH2 (humoral/antibody reaction)
T helper cells (TH)
interleukins o TH17 (inflammation)
• Express surface markers CD2,
CD3, CD4
• Requires interaction with TH to become • Express CD2, CD3 and CD8 on their
Cytotoxic T cells (TC) activated surfaces
• Kills virus-infected and cancer cells
• Suppress immune responsiveness to self- • Express CD4 and FOXP3 markers
antigens (autoimmunity)
Regulatory T cells (TREG)
• Switch off the response when antigen is
completely removed
• Develop from activated t cells
Memory T cells • Rapid reaction force to subsequent ---
encounter with same antigen
γδ T cells • Epithelium of the GI tract (GALT) • CD8 positive

5.1.2. B Lymphocytes [WYB]

• Produced and mature in the bone marrow

• Stimulated B cells further mature into plasma cells
o Usually synthesize large amounts of antibodies in the lymph nodes
IgG IgM IgA IgD IgE
Pentamer Dimer with J Chain Monomer
Monomer and Secretory Monomer
Component
Structure

Antibody % in
75-85% 5-10% 10-15% 0.001% 0.002%
the Plasma
Presence in
Bound to the
sites other than Fetal circulation B lymphocyte
Secretions (saliva, Surface of B surface of mast
blood, CT, and in pregnant surface (as a
milk, tears, etc.) lymphocytes cells and
Lymphoid women monomer)
basophils
Organs

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First antibody Antigen receptor

Activates produced in triggering initial B Destroys
Known phagocytosis, initial immune cell activation; parasitic worms
Protects mucosae
Functions neutralizes response; Part of the T cell and participates
antigens activates – B cell receptor in allergies
complement complex

✂ Additional Indicates passive Indicates

Notes immunity against recent/active - - -
(Batch 2025 Trans,
certain infections infection
Dr. Mupas-Uy)
CT – Connective Tissue

Primary Immune Response

• Antigen is encountered for the first time
• Undergoes Mitotic Division
o Clones of cells able to synthesize
immunoglobulin of the same antigen specificity
Few cells become Memory
MOST mature into B Cells, small long-lived
Plasma Cells circulating lymphocytes
(secretes IgM) • Able to respond quickly to
any subsequent challenge
with the same antigen

Secondary Immune Response

• Initiated by Memory B cells
• Antibody Production
o More rapid with much greater magnitude
o Produces IgG rather than IgM

5.2. Antigen Presenting Cells [WYB]

• Vital for Lymphocyte Activation Fig. 5.2.1. Gene regulatory networks
• Macrophages, Dendritic Cells, and B
lymphocytes
• Functions
o Patrol the body surfaces
o Phagocytose invading pathogens
o Reside in the lymph nodes and in the peripheral
tissues
Macrophages • Tissue histiocytes
• Paracortical area of lymph node
• Interdigitating Cells (in Thymus)
• Langerhan’s Cells (in Skin)
Dendritic Cells • Follicular Dendritic Cells (in
Germinal Center of Lymph Nodes) Fig. 5.2.2. EM of a Dendritic Cell
o Able to bind antigen-antibody • Exhibits long branched cytoplasmic extensions
complexes to their surface • Interacts with several lymphocytes in its periphery
without prior processing
• Interact with ‘unprocessed
B Cells
antigens’ 6. IMMUNE RESPONSE [WYB]
6.1. Recognition of Antigen [WYB]
• T and B cells carry antigen receptors on their surface
o T Cell Receptors (TCR)
o B Cell Receptors (BCR)

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• Random rearrangement of the variable region gives

rise to receptors with a staggering range of receptor
binding sites
o Each individual T or B cell has specificity for
ONLY ONE ANTIGEN

6.2. Activation of Immune Response [WYB]

6.2.1. T Cell Activation

• Dependent on Antigen Presenting Cells and

Major Histocompatibility Complex (MHC)
Fig. 6.2.2.1. B Cell activation
• Expressed in ALL nucleated cells
o Except for mature RBC and
platelets
MHC 6.3. Generation of Effector Mechanisms [WYB]
• INTRINSIC antigen
Class I
o Viral infected cells
6.3.1. Antibody-mediated Antigen Elimination
o Tumor cells
• Recognized by Cytotoxic T cells Binding to Viral Surface Antigens
• Expressed in Antigen Presenting cells • Antibodies block entry of microorganisms
o Dendritic Cells
o Macrophages
o B cells
• EXTRINSIC Antigen
o Bacterial cells
• Antigen is processed by an APC and
MHC
broken down into short peptides
Class II
o Processed antigen is then bound to
a MHC II molecule
o Complex is incorporated into the
cell membrane
o Bound antigenic peptide is exposed
to ECF
• Recognized by T Helper Cells

Fig. 6.3.1.1. Binding to Viral Surface Antigens

Activate the Complement System

• Antigen-Antibody complexes activate the
complement cascade and induce the Membrane
Attack Complex

Fig. 6.2.1.1. MHC Class I Fig. 6.2.1.2. MHC Class II

6.2.2. B Cell Activation

• ‘Unprocessed’ Antigens
o Usually presented by a Follicular Dendritic Cell
within a lymphoid follicle
• B Cells recognize the antigen by means of the BCR
(Surface Ig)
o Requires co-activation with a T-Helper Cell
• Activation of a B cells without T cell help
o Protein or polysaccharide antigen
o Repeating Chemical Structure
▪ Pneumococcus sp.
▪ T cell Independent antigens
Fig. 6.3.1.2. Activate the Complement System

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Opsonization o Antibody Dependent Cytotoxic

• Bound antibodies with or without complement Cells
opsonizes microorganisms
• Facilitates its phagocytosis by neutrophils and
macrophages

Fig. 6.3.2.1. Cell-mediated cytotoxicity

6.3.3. Activation of Macrophages

• Certain types of organisms like mycobacteria

activate Th1 cells to secrete cytokines that activate
Fig. 6.3.1.3. Opsonization
macrophages
Antibody Dependent Cell Cytotoxicity • More efficient at killing phagocytosed organisms
• Ends in the apoptosis of the cells (tumor cells) • Chronic Granulomatous
Type IV
Inflammation
Hypersensitivity
• Tuberculosis

Fig. 6.3.3.1. Granuloma

Fig. 6.3.1.4. Antibody Dependent Cell Cytotoxicity

Inactivation of Toxins
• Antibodies bind to toxins and facilitates their removal 6.4. Termination of Immune Response [WYB]
through phagocytosis • Prevents damage to normal tissues
• Prevents autoimmunity
• Mechanisms:
o Complete Removal of Antigens
o Short life span of Plasma cells
▪ Renders the immune response time bound
o Activities of Regulatory T cells

7. LYMPHOID ORGANS [WYB](C16)

Major Lymphoid Organ Classification
Fig. 6.3.1.5. Inactivation of Toxins Primary Secondary
• Spleen
• Bone Marrow
• Lymph nodes
6.3.2. Cell-mediated Cytotoxicity • Thymus
• MALT
• Destruction of abnormal cells by:
Apoptosis o Cytotoxic T Cells
o Natural Killer Cells

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7.1. Bone Marrow(C16) 7.2.1 Thymic Lobule

• Home of lymphoid stem cells which give rise to: • Highly cellular outer cortex
o Immature T cells • Less cellular central medulla
o Immature B cells
• Site of maturation of B cells

Fig. 7.2.1.1 Thymic lobule

Cortex – highly cellular; Medulla – less cellular
Fig. 7.1.1. Bone Marrow

7.2.2. Blood Thymus Barrier

7.2. Thymus(C16)
• Flattened lymphoid organ • Epithelial cells forming sheaths around blood vessels
• Located in the upper anterior mediastinum and lower • Serves as an impenetrable barrier
part of the neck o Prevent the entry of antigenic material into the
thymic parenchyma
• Most active during childhood
• Undergoes slow involution
o Difficult to differentiate from adipose tissue
macroscopically in middle-aged or older adults
• Single organ subdivided into numerous fine
lobules

Fig. 7.2.1. Thymus

✎ THYMUS (p. 204) Fig. 7.2.2.1. Blood Thymus Barrier

The epithelial cells of the thymus provide a mechanical
supporting framework for the lymphocyte population.
• Cortical epithelial cells also promote T cell differentiation 7.2.3. Infant vs. Adult Thymus
and proliferation. Furthermore, the epithelial cells secrete
a number of different hormones that regulate T cell THYMUS
maturation and proliferation within the thymus and in other IN INFANTS IN ADULTS
lymphoid organs and tissues.
• The inner surfaces of the thymic capsule and septa are
invested by a continuous layer of thymic epithelial cells
resting on a basement membrane.
• The epithelium also forms sheaths around the blood
vessels, creating a barrier to the entry of antigenic material
into the thymic parenchyma. This is known as the blood-
thymus barrier.

• In the process of
• Well developed involution
• Involves 2 processes:
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o Fatty infiltration
o Lymphocyte
depletion
• Described as small islands
of lymphoid tissues in a
sea of fat
✎ INFANT THYMUS (p. 204-205)
• Lobulated organ invested by a loose collagenous
capsule C from which interlobular septa S containing
blood vessels radiate into the substance of the organ.
• The thymic tissue is divided into two distinct zones, a
deeply basophilic outer cortex Cx and an inner
eosinophilic medulla M

Fig. 7.2.5.1. Development of self-tolerance through

negative selection & apoptosis

7.2.6. Thymic Cortex

THYMIC CORTEX
Outer Cortex Deep Cortex
• Presence of • Maturing T cells
lymphoblasts
• Divide by mitosis
• Thymic nurse cells –
specialized epithelial
7.2.4. Types of Involution cells in the outer
cortex
Acute Thymic Involution Stress Involution
• In response to severe
• Characterized
disease and metabolic stress
by greatly
• Associated with:
increased
o Pregnancy lymphocyte
o Lactation
death
o Infection
• Mediated by
o Surgery
high levels of
o Malnutrition
corticosteroids
o Malignancy

7.2.5 Functions of the Thymus

• Production of mature Th and Tc cells

• Proliferation of clones of mature naïve T cells
• Immunologic self-tolerance Fig. 7.2.6-1 Thymic Cortex
• Secretion of various polypeptides: thymulin,
thymopoietin, thymosins • T cell maturation is promoted by
o regulate T cell maturation, proliferation and interaction with thymic nurse cells,
function within the thymus and peripheral which are specialized epithelial cells
Thymic
lymphoid tissues found in the outer cortex
Nurse
o interact with other lymphoid organs in the • During this process, T cell receptor
Cells
regulation of inflammation genes are rearranged and the cells
acquire the surface markers of mature
helper and cytotoxic T cells
SURFACE MARKERS
T Helper Cells Cytotoxic T Cells
CD3 & CD4 CD3 & CD8
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• Thymocytes failing to make these

adjustments die by apoptosis
Apoptosis
o Taken up by pale-stained
macrophages

Fig. 7.2.7-2 Interdigitating Dendritic Cell

Fig. 7.2.6-2 TCR Gene Rearrangement 7.2.8. Staining and Immunohistochemistry

Standard H&E Stain

7.2.7. Thymic Medulla • The dense population of lymphocytes in the thymic
parenchyma make the epithelial cells and
• Where the mature thymocytes live macrophages generally difficult to visualize in the
• Dominant histological feature: robust epithelial standard H&E
component composed of epithelial cells having large • Fortunately, these cells can be highlighted by
pale-stained nuclei, eosinophilic cytoplasm and immunohistochemical techniques
prominent basement membranes
• Lamellated structures
• First appear in fetal life – increase in
number and size thereafter
Hassall • Formed from groups of keratinized
Corpuscles epithelial cells, with fragments of
debris at their center
o Represent a degenerative
phenomenon

Fig. 7.2.8.1. Standard H&E Stain

Immunohistochemistry
• Demonstrates the extensive network of epithelial
cells in the cortex & the more rugged epithelial
network of the medulla

Fig. 7.2.7-1 Thymic Medulla

Ep – Epithelial cells; HC – Hassall Corpuscles

• Specialized dendritic cells found

in the thymic medulla
• Express high levels of both class
I and II MHC proteins
Thymic • Perform clonal deletion or
Interdigitating negative selection by presenting
Cells self-antigens to maturing T cells
o Self-reactive T cells are
obliterated by apoptosis
→ prevent development
Fig. 7.2.8.2. Thymus tissue stained using antibody to cytokeratin
of autoimmunity AE1/3 (Pancytokeratin)
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Fig. 8.2. Acute inflammatory response

7.3.1. Functional Compartments

Fig. 7.2.8.3. Thymic tissue stained with antibody to CD 68 1 STROMA OR CELLULAR COMPARTMENT
(strongly expressed by macrophages and dendritic cells) • Made up of lymphocytes and APCs

7.3. LYMPH NODES

• Bean-shaped, encapsulated, highly organized
structures
• Human body has 450 lymph nodes
o Grouped in areas where the lymphatics
converge to form larger trunks: in the neck,
axillae, groins, lung hila, mesentery of the
bowel, and paraaortic areas Fig. 7.3.1-1 Stromal Compartment
• Process antigens from the lymph 2 SINUS COMPARTMENT
• Primary site to stimulate an immune response to • Consists of the:
antigens in the lymph o Subscapular sinus
• Bring together antigens, potentially reactive o Trabecular sinus
lymphocytes, and APCs o Medullary sinus
• Provide the best environment to stimulate an
adaptive immune response

Fig. 7.3.1-2 Sinus Compartment

3 VASCULAR COMPARTMENT
• Delivers lymphocytes to the lymph node along with
Fig. 8.1. Lymph node
the usual nutrients

Acute Inflammatory Response

• Increases the flow of lymph by flooding the
infected/damaged tissue with extracellular fluid
o Increases the chances of antigen recognition &
processing

Fig. 7.3.1-3 Vascular Compartment

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7.3.2. Structure and Vascular Organization Medulla

• Inner central area
• Collagenous •
Capsule Less cellular than the cortex
• Trabecular extensions • Dominant feature of the medulla
• Where efferent lymphatics drain efferent • Network of broad interconnected
lymph from the lymph node lymphatic channels
Hilum • Site of entry of the artery bringing blood • Converge upon the hilum (H) in the
to the lymph node and the exit of the vein concavity of the node
leaving the node •
Medullary Lymph drains into the efferent
• Highly cellular outer part of the lymph
Cortex Sinuses lymphatic vessels which drain into
node more proximal lymph nodes that then
join the blood stream via the thoracic
duct or right lymphatic duct
o Process done to facilitate the
exposure of a large number
of lymphocytes to antigens
in the lymph
Medullary • Extensions of the cortical cell mass
Cords

Fig. 7.3.2-1 Lymph Node Structure

Superficial Cortex
• Where B cells live
• Contains a number of dense cellular aggregations
called ‘follicles’
Primary follicles Secondary follicles
• No germinal center • With germinal centers Fig. 7.3.2-4 Medulla
• Inactive (pale-stained) Medullary Sinuses (MC); Medullary Cords (MC)
• Active
Vascular System
• Main route of entry of lymphocytes into the node
• Supplies the nodes with its metabolic requirements
• Within the paracortex
High
• Lined by cuboidal epithelium
Fig. 7.3.2-2 Superficial Cortex with Primary (PF) & Endothelial
specialized for the exit of
Secondary (SF) Follicles Venules (HEV)
lymphocytes
Paracortex • Specific complimentary adhesion
• Deeper cortex molecules found in the HEV of
• Where T cells live different lymph node groups
• Densely cellular like the outer cortex but has a more • Corresponding binding molecules
homogeneous appearance on the lymphocytes
Addressins
• T lymphocytes interact with APCs in this area and o Recognition of exit sites
undergo a similar process of activation and clonal require the presence of
expansion addressins on the surface of
both the endothelial cells and
lymphocytes

Fig. 7.3.2-3 Paracortex

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7.3.4. Reticular Network

Capsule
• Main structural support
and
• Extend into the node
Trabeculae
• Dense in the cortex
o Sparse in the follicular areas
• Fine meshwork that extend
throughout the node from the capsule
and trabeculae
• Provide a supporting framework for
Fig. 7.3.2.5. Vascular system Reticulin the mass of lymphocytes and
Fibers accessory cells within the stroma
✎ LYMPH NODE ENLARGEMENT (p. 207) • Stained blackish brown by the
• Due to viral or bacterial infections reticulin method
o Increased flow of lymph to lymph nodes when o The light brown-stained
triggering an adaptive immune response → lymph
node enlargement
structures in the follicles are
o Seen in pharyngitis which may lead to the enlargement lymphocytes and not reticulin
of the tonsils (tonsillitis) fibers
• Due to malignant tumors (lymphomas & leukemias) • Structural skeleton
o Increased flow of lymph possibly due to necrosis of the o Collagen framework
Fibroblastic
tumor o Similar to a sponge
✎ METASTASIS Reticular
• Facilitate Ag presentation
• Cells
Spread of tumor to lymph nodes • Have a role in bringing together
• Important for staging a tumor dendritic cells and T cells
o Local lymph nodes are sampled or removed to assess
for presence of tumor deposits
o Routine in assessment of breast and colonic
carcinomas
✎CHRONIC GRANULOMATOUS INFLAMMATION
• Infective organisms directly invade and grow in the lymph
node
• Such infections include:
o Mycobacterium tuberculosis (TB)
o Mycobacteria other than tuberculosis (MOTT)
o Non-tuberculous Mycobacteria (NTM)
o Toxoplasma gondii

7.3.3. Flow of Lymph

• Afferent lymphatic vessels Fig. 7.3.4.1. Lymphoid tissue stained using reticulin method
• Subscapular sinus
• Cortical and trabecular sinus
• Medullary sinus 7.3.5. Conduit System
• Efferent lymphatic vessels
• Component of the collagenous skeleton
o Specialized collagen fibers wrapped in basement
membrane
o Covered by Fibroblastic reticular cells
• Run from Subscapular Sinus to HEV
• Thought to carry soluble antigens and cytokines into
the lymph node parenchyma
• They also carry fluid from the lymph back to the
bloodstream. Hence, the lymph leaving the node is
more concentrated than the lymph entering it

Fig. 7.4.1. Flow of Lymph

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Fig. 7.3.5.1. Conduit system

7.3.6. Distribution of T and B Lymphocytes

Fig.7.3.6.2. Micrograph showing lymph node distribution of T and

B Lymphocytes
Aggregated of B cells in the germinal center stained by
CD20

Fig.7.3.6.1 Micrograph showing lymph node distribution of T and

B Lymphocytes
Unstained secondary follicle with a germinal center T cell
located in the paracortex highlighted by CD3.

Fig.7.6.3. Micrograph showing lymph node distribution of T and B

Lymphocytes
T cells located in the paracortex highlighted by CD3

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7.3.7 Paracortex – High Endothelial Venules • From resting B cells

• Large mitotically active cells with
round nuclei that are found in the
Centroblasts darker zone of the germinal
center, closer to the medulla
• Further differentiate into
centrocytes
• Found in the paler zone of the
germinal center towards the
lymph node capsule
• Are of variable size and have
folded, irregular, or cleaved
Centrocytes nuclei
• Absent mitotic figures
• Go through further cycles of cell
Fig.7.3.7.1 Paracortical zone division to produce either
(Left) Unstained paracortical zone with circulating T cells immunoblasts or Memory B
entering the lymph node through the walls of the high Cells
endothelial venules (easier seen in the right picture).

Fig.7.3.7.2. Paracortical zone

(Right) showing two HEVs surrounded by T lymphocytes
stained with CD3

7.3.8. Germinal Center Cells

Fig.7.3.8.1. Germinal center
Germinal Centre of Secondary Lymphoid Follicle (Yellow arrow) – Centroblasts
• Mainly actively dividing B cells (Blue arrow) – Centrocytes
• Resting B cells enter the lymph node via the HEV (Red arrow) – Macrophages
Blast Transformation • Resemble small lymphocytes
• Antigen encounter • Take up residence in the
• Activation Memory B Cells mantle zone or may join the
recirculating pool of small
lymphocytes
• Move to the medullary cords
were they complete their
differentiation into plasma
Immunoblasts cells
• Plasma cells – capable of
secreting large amounts of
antibodies

Fig. 7.3.8.1. Lymphocyte activation

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Fig.7.3.8.4. Folicular Dendritic Cells

Tingible Body Macrophages

Fig.7.3.8.2 Centrocyte differentiation
• Tingible = stainable
Somatic Hypermutation • Easily seen in routine sections in active germinal
• Centroblasts undergo increased mutation of the centres
immunoglobulin genes and performs class switching • Contain within their cytoplasm numerous apoptotic
• Creating further variations in immunoglobulin bodies
structure o B lymphocytes
o Unsuccessful in generating high-affinity
antibodies

Fig.7.3.8.3. Somatic Hypermutation Fig.7.3.8.5. Tingible Body Macrophages

Follicular Dendritic Cells

• Major antigen presenting cells of the lymphoid 7.3.9. Medullary Cord and Sinuses
follicles
• Found in all areas of the germinal centre, form a MEDULLA
meshwork mantle zone and in primary follicles • Branching medullary cords separated by irregular
• Retain antigen on the urgace for many months medullary sinuses
• Present unprocessed antigen to B cells • Major cell types in the
• Maintain the activity of Memory cells medullary cords
• Stimulate a primary immune response • Final stage of maturation of B
Plasma Cells &
cells to Plasma cells (in the
Plasmablasts
medulla)
(precursor)
o Synthesize antibody that is
carried to the general
circulation

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o Some plasmablasts Endothelial • Lining the sinus

mugrate to the peripheral cells
tissues

Fig.7.3.10.1. Subscapular sinus at higher magnification

Traversed by fine reticulin strands and endothelial cell
Fig.7.3.9.1. Medullary Cord (MC) and Sinuses (MS)

• Packed with plasmablasts and

Medullary Cord
plasma cells (P)
• Contains mainly macrophages
Medullary Sinus
(M) or “sinus histiocytes”

Fig. 7.3.10.2. EM shows structure of the Subscapular Sinus

Yellow arrow – Supporting network of reticular fibers

7.4. Mucosa-Associated Lymphoid Tissues (MALT)

Fig. 7.3.9.2. Micrograph showing a high magnification view of a
medullary cord and sinus
In the center the medullary cord is packed with plasmablasts and
plasma cells. While the sinus in fbevperiphery contains mainly
macrophages
Total Mass of Mucosal Lymphoid Tissues
• Gastrointesinal Tract (Gut-Associated Lymphoid
Tissue, GALT)
• Respiratory Tract (Bronchial-Associated Lymphoid
Tissue, BALT)
• Oropharynx (Waldeyer Ring/NALT)

7.3.10. Subscapular Sinus

• Provide support for large sinus

macrophages (M)
Fine Reticulin
• These sinus histiocytes filter
Strands
antigen and other debris from
afferent lymph

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o Secretory IgA protects against pathogens in the

gut lumen before they breach the tissues
Antigen Processing and Lymphocyte Circulation
MALT acts as an integrated unit with a separate route of
lymphocyte circulation

Fig. 7.4.1. Mucosal lymphoid tissue distribution

MALT may be arranged either as:

• Lamina propria
Diffuse
• Mainly T lymphocytes with a
Population of
small number of B cells as well
Cells
as Plasma cells Fig. 7.4.3. Antigen processing and lymphocyte circulation
Non- 1
Antigen encountered in the epithelium
Encapsulated/ • Carried to local MALT tissue
• Tonsils 2 Lymphocyte Stimulation
Hemicapsulated
• Peyer’s patches
organized Lymphocyte Migration
3
aggregations • Regional (Mesenteric) lymph nodes
• Lymphocytes and plasma cells Clonal Expansion
which become highly functional • Production of effector cells
Breast 4
during breastfeeding and • Back to the mucosa
infections o Addressins

7.4.1. Tonsillar Ring of Waldeyer

• Organized masses of lymphoid tissues

o Palatine Tonsil
o Pharyngeal Tonsil
o Lingual Tonsil
o Tubal Tonsil (Adenoids)

Fig. 7.4.2. Secretory IgA

• All antibody classes are produced in MALT

• Predominantly IgA
o Bound to a carbohydrate moiety that renders it
resistant to proteolytic enzymes
Fig. 7.4.1.1. Tonsils
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Palatine Tonsil
• Stratified Squamous Epithelium (E) on the luminal
side.
• Epithelium has blind-ended invaginations known as
Tonsillar Crypts (C)
• Base of the tonsil is separated by the underlying
muscle by a dense and collagenous Hemicapsule
(Cap)
• Tonsillar parenchyma contains numerous lymphoid
Follicles (F) with Germinal centers like those found in
the lymph nodes
• Function in a manner analogous to the lymph node
• Particulate matter or bacteria enter the crypts and Fig. 7.4.1.3.. Tubal tonsil
passed to the follicles by transcytosis
• Immune response initiated
7.4.2. Gut Associated Lymphoid Tissues (GALT)

Organized lymphoid tissues found in all parts of the

GIT EXCEPT THE STOMACH
• Largest lymphoid aggregates in the
GALT
Peyer’s • Located in the mucosa
Patches • Bulges dome-like into the gut lumen
of the SI • Few villi or no villi
• Least numerous in the duodenum
• Most prominent in the terminal ileum
• Low cuboidal epithelial cells that
M Cells
perform transcytosis of antigens

Fig. 7.4.1.2. Palatine tonsils

Fig. 7.4.2.1. GALT

Galt is prominent in the Appendix

• Appendiceal mucosa with a lymphoid follicle
exhibiting an active germinal center

Fig. 7.3.12.3. Palatine tonsils

Tubal Tonsil (Adenoid)

• Reflect a respiratory type of epithelium

Fig. 7.4.2.2. GALT in appendix

• Brown-stained aggregates of B-cells using the
antibody CD20
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Fig. 7.4.2.3. GALT stained with CD20

7.4.3. Bronchial Associated Lymphoid Tissue (BALT)

• Part of MALT
Fig. 7.5.1. Spleen
• Consists of lymphoid follicles in the lungs and
bronchus Functions
• Efficient priming site of the mucosal and systemic • Production of immunological responses against
immune response (adaptive B-cell and T-cell bloodborne antigens
responses) directed against airborne antigens • Performs the same functions for blood that lymph
nodes perform for lymph
• Removal of particulate matter and aged or defective
erythrocytes
• Recycles iron to the bone marrow
• Hematopoiesis is the normal fetus and adults with
certain diseases

7.5.1. Components of the Spleen

Splenic Parenchyma
Red Pulp White Pulp
• Dark red • Small macroscopically
parenchyma visible white nodules
• Vascular Area • Lymphoid Area

Fig. 7.4.3.1. BALT

7.5. Spleen
• Large lymphoid organ in the LUQ
• Blood Supply: Splenic Artery
• Drainage: Splenic Vein into the hepatic portal system

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• Central artery branches out in right angles as

penicillary arteries

Fig. 7.5.1.1. Gross section of spleen and splenic parenchyma

showing the white pulp (L) and red pulp (R)

Capsule Trabeculae
• Thin fibroelastic • Extensions of the
tissue capsule into the
• Outer surface parenchyma
covering of
mesothelium • End up in the sheathed capillaries
• Easily ruptured with
blunt trauma which
causes the spleen to
bleed, sometimes
leading to
splenectomy

Fig. 7.5.1.2. Parts of the splenic parenchyma
7.5.2. Splenic Circulation
• Blood Enters the Splenic Artery
Open Circulation
• Starts with the sheathed capillaries which are blind-
ended vessels not lined with endothelial cells, but
instead surrounded by macrophages
• Once in the splenic parenchyma, particulate matter or
aged erythrocytes are removed by the macrophages
• The recognition of defective RBCs is probably based
on diminished deformability, but immunological
mechanisms may also be involved
• The blood cells move out from the extravascular
space by squeezing through the walls of the sinuses
to eventually drain to the splenic vein
• This is part of the antigen-filtering mechanism of the
spleen where macrophages act as the filtration
agents in the extravascular space

• Flows down the central artery surrounded by the

periarteriolar lymphoid sheath (PALS) Fig. 7.4.2.1. Open circulation of the spleen

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7.4.3. Parenchyma Components

Non-Filtering Areas (NFA)

• The non-filtering areas of the parenchyma (boxed in
yellow) are areas devoid of capillaries and contain
mainly T and B lymphocytes and macrophages

• Lymphocytes exit through these adjacent sinuses

(pointed by the blue arrows) with endothelial linings
similar with high-endothelial venules of lymph nodes
• These non-filtering areas of the red pulp parenchyma
is also a functional part of the splenic lymphoid tissue
Fig. 7.4.3.1. Red pulp venous sinus
Staining
Reticulin Method CD 68
• Highlights the reticular • Highlights the
network of the spleen presence of splenic
• Basement membranes macrophages in the
show greatest red pulp parenchyma
concentration of
• Zone of the red pulp immediately surrounding the
Reticulin fibers
white pulp (red box) is called the perilymphoid red
pulp.
• Function unclear, but sluggish blood flow may be to
enhance interaction of blood cells, antigens, and
antibodies

White Pulp – T and B Cell Zones

• T cell Type & B cell Type
• Both make up 5-20% of the total splenic mass
• Functions similar to the paracortex and superficial
Red Pulp – Venous Sinuses
cortex of lymph nodes, respectively
• Red pulp is permeated by broad interconnected
venous sinuses
• 3D structure is analogous to Swiss cheese
o Holes: Sinuses
o Cheese: Parenchyma
• Lined by elongates spindle-shaped cells
• Likened to a tall wooden barrel with both ends open
o Wooden staves: Endothelial cells
• Slits occur within the endothelial cells with a
discontinuous basement membrane over the slits T Cell Zone
• Viable erythrocytes squeeze between the stave cells • Form an eccentric cylindrical sheath containing small
to reach the venous sinuses lymphocytes mainly of the T helper subset around a
central artery (yellow arrow)
• Equivalent of the periarteriolar lymphoid sheath in
animals

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✎ TUMORS OF THE IMMUNE SYSTEM (p. 219)

• As a group they are called lymphomas (involving solid
organs) and leukemias (involving the blood) and exist in
many forms with many characteristic clinical presentations,
occurring in virtually all age groups
• There are no truly benign tumors of lymphoid tissues
LYMPHOMA
• May be systemic or may be localized to a particular
lymphoid organ or to non-lymphoid organs such as the skin
B Cell Zone or brain.
• B cells form follicles that exhibit germinal centers • They may occur in otherwise healthy individuals, in
similarly seen in lymph node especially in young immunosuppressed people such as AIDS patients or organ
people transplant recipients or in certain infections.
• In the follicle periphery is a narrow zone of small • For instance, patients with lymphoma of the stomach
(MALT lymphoma or MALToma) almost always have
lymphocytes called the mantle zone
infection of the stomach with Helicobacter pylori, a
• Beyond that is a broader marginal zone composed of bacterium also associated with gastritis, peptic ulcer and
less dense but medium sized lymphocytes gastric adenocarcinoma.
o Interestingly, eradication of the infection may bring
about resolution of the lymphoma, at least in the early
stages.
• Enlargement of some or all lymph nodes
(lymphadenopathy)
Clinical • Enlargement of the spleen
Presentations (splenomegaly) and liver
(hepatomegaly)
• Fever, weight loss and malaise
Staining
Non- • Composed of malignant B lymphocytes
CD 3 CD 20 may have a follicular (nodular)
• T cells stained with CD3 • B cells stained with Hodgkin
architecture, recapitulating normal
and found along the CD 20 forming a Lymphoma lymphoid follicle formation
sides of the central primary follicle
artery
9. SLIDE REVIEW
9.1. Thymus

8. TUMORS OF THE IMMUNE SYSTEM

Lymphomas (involving solid organs)
• Systemic or Localised
• Lymphoid or non-lymphoid organs
• Healthy individuals
• Immunosuppressed people (AIDS, organ recipients)
• Infections (H. pylori) • Single organ sub-divided into numerous fine lobules
• Thymic lobule has highly cellular outer cortex and
less cellular inner medulla
o Separated by fibrous septations

• Notice the effacement of the lymphoid architecture on

the normal slide, replaced with a papillation of
neoplastic small round monomorphic cells in
lymphoma

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9.1.1. Infant Thymus • The lamellated corpuscles formed from groups of

keratinized epithelial cells [probably] represent
degenerative phenomenon
• The dense population of lymphocytes in thymic
parenchyma make the epithelial cells and
macrophages difficult to visualize; This is why we use
immunohistochemical techniques

9.1.4. Immunohistochemistry

• Thymus is well-developed in infants

• In adults, involution happens; This involves fatty
infiltration and leukocyte depletion
o Small island of lymphoid tissues in a sea of fat
STAINS
Antibody to Cytokeratin
Antibody to CD68
9.1.2. Thymic Cortex AE1/3
Demonstrates extensive
network of epithelial cells
Highlights macrophages
in the cortex and
and some dendritic cells
epithelial framework of
medulla

9.2. Lymph Node (LWJ)

• In the outer cortex, large lymphocytes or

lymphoblasts divide by mitosis to produce clones of
smaller mature T cells towards medulla
• These undergo further maturation as they move
deeper towards the center in the inner medulla

• Stromal compartment – packed with lymphocytes

and antigen-presenting cells
9.1.3. Thymic Medulla

9.2.1. Lymph Node Cortex

SUPERFICIAL CORTEX

• The dominant histologic feature of the thymic medulla

is the robust epithelial component
• Composed of: • More cellular than inner/cortical medulla
o Epithelial cells with large pale-stained nuclei • Where B cells live
o Eosinophilic cytoplasm • Contains dense cellular aggregation called follicles
o Prominent basement membrane • Secondary follicles - active (pale-stained germinal
centers
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DEEP CORTEX/PARACORTEX

• Where T-cells live

• Densely cellular like outer cortex but with more
homogenous staining appearance
• T-lymphocytes interact with antigen-presenting cells
in this area

• Central part: Medullary cord is packed with

9.2.2. Lymph Node Medulla plasmoblasts and plasma cells
• Sinus in periphery: contains mainly macrophages
SUBSCAPULAR SINUSES

• DOMINANT FEATURE: Extensive network of broad

interconnected lymphatic channels called medullary
sinus • Traversed by fine reticulin strands that provide
o Lymphatic sinuses converge in hilum at the support for histiocytes (large sinus macrophages)
concavity of node • Macrophages filter antigen and other debris from the
afferent lymph
MEDULLARY CORDS AND SINUSES • Endothelial cell lining the sinus can also be identified

• Structures of subscapular sinus

o Highlighted by sinus macrophage
o Endothelial cell lining
o Supported network of reticular fibers pointed by
• Micrograph: Structure of lymph node medulla with the yellow arrow
branching medullary chords separated by irregular
medullary sinuses
• Plasma cells and precursors (Plasmoblasts)
o major cell types in medullary cords
• In medulla, B lymphocytes complete their final stage
of maturation

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9.2.3. Structural Components of Lymph Node
• Left Picture: Unstained paracortical zone with
circulating T cells entering lymph node through walls
of high endothelial venules
• Right Picture: 2 high endothelial venules and exiting
lymphocytes from vascular channels

• Yellow Arrow: Centroblast

• Main structural support for the lymph node: derived
• Blue arrow: Centrocyte
from collagenous capsule and trabeculae
• Red Arrow: Macrophage
o Extend from capsule to lymph node parenchyma:
trabecular extensions
o A fine meshwork of reticulin fibers extend
throughout the node to provide supporting 9.2.5. Lymphocytes
framework
• Stained blackish brown by reticulin stain
• Dense in the cortex except for follicular
Reticular
areas (relatively sparse)
Network
• Note: the light brown-stained structures
are lymphocytes and not reticulin fibers

• Lymphocytes have round to ovoid nucleus

o Occupies 90% of cell volume
• Surrounded with a thin rim of basophilic or bluish
• Left Picture: Unstained secondary follicle with cytoplasm
germinal center
• Middle Picture: CD20 B cells straining
o Found in secondary follicle 9.2.6. Dendritic Cells
• Right Picture: CD3 staining of T cells in paracortex
Lymphatic sinuses converge in hilum at the concavity
of node

9.2.4. Paracotical Zone

• Interacts with several lymphocytes in its periphery

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• Found in tonsillar
Lymphoid
F parenchyma; Contain
Follicles
germinal centers

• Follicular dendritic cells

• Antibody CD21 highlights presence of follicular
dendritic cells at the lymphoid germinal center

9.2.7. Lymph Flow on Lymph Node

9.2.12. GALT

Peyer’s Patches
Description Largest lymphoid aggregates in GALT
Location Small intestine
Villi Few or no villi overlying it
• Lymphoid follicle like seen in lymph
nodes
Contains • Prominent active germinal center (GC)
• Mantle zone of small resting
lymphocytes
M Cells
Description Low cuboidal epithelial cells
• Afferent lymphatics → goes down to subscapular Location Scattered among epithelial cells
sinus → cortical and trabecular sinus moving Function Perform transcytosis of antigens to the
downwards to medullary sinus → efferent lymphatics subepithelial lymphoid cells
• The presence of conduit system – transports water
and electrolytes from subscapular sinus straight to
high endothelial venules
• Efferent lymph is more concentrated than the afferent
lymph

9.3. Mucosa-Associated Lymphoid Tissue

9.3.1. Palatine Tonsil

• Luminal Surface: Stratified

E Epithelium
squamous epithelium
• Blind-ended; Formed by
Tonsillar
C invaginations of the
Crypts
epithelium
• Dense, collagenous;
Cap Hemicapsule Separates base of tonsil
from underlying muscle Peyer’s Patch in Ileum

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Appendiceal Mucosa CD20 (IHC)

Peyer’s patches are Brown-stained aggregates
prominent in the appendix; of B cells using antibody to
Micrograph contains a CD20
lymphoid follicle exhibiting
active germinal center

3 Penicillar arteries

Left: Appendiceal Mucosa

Right: Staining of B Cells (CD20 [IHC])

9.4. Spleen (LWJ)

• Thin, fibroelastic
Capsule • Easily ruptures in blunt abdominal
trauma
• Extend from capsule into the 4 Sheathed capillaries
Trabeculae
parenchyma

Open Circulation
Blind ended vessels lined by
Sheathed
macrophages; 1st part of antigen
Capillaries filtering system of the spleen
Act as filtration agents in extravascular
Macrophages
space

9.4.1. Splenic Circulation

Splenic artery
1
Splenic circulation starts when blood enters here

Central artery
2 Surrounded by periarteriolar lymphoid sheath
Branch out in right angles to form penicillar a.

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9.4.2. Red Pulp Venous Sinuses B-Cell Zone

Stave • Elongated spindle shaped endothelial

Cells cells that line venous sinuses in the red
pulp
Blood • Squeeze between endothelial cells to
Cells reach venous sinuses

• Exhibit germinal centers like those

Follicles
in lymph nodes

REFERENCES
• Young, B., Woodford, P., & O’Dowd, G. (2013). Wheater’s
functional histology (6th ed.). Churchill Livingstone.
• Lecture recording of Dr. Luna
• Upper batch trans
9.4.3. White Pulp
TL: HCZ (1B)
T-Cell Zone
FREEDOM WALL

• Eccentric
• Formed around the central artery
Cylindrical • Equivalent to periarteriolar lymphoid
Sheath sheath in animals
Small lymphocytes (mostly T
Contains
helper)

HISTOLOGY 29