Basic Immunololgy

 Introduction to Immunology
 Innate immunity
 Cell Biology
 Complement system
 Antibody dependent responses
 Cell mediated immune responses
 Immunoregulatory mechanisms
 INTRODUCTION TO IMMUNOLOGY
Immunology
 Biochemical study of body defenses against

parasites and microbes (bacteria, virus, fungi

and parasites).
Immunology important in
 Biomedical, clinical, dentistry, pharmacy and

basic sciences
 Evolutionary trends from innate to adaptive

immunity
 Immunodiagnostics in the treatment and

management of patients including underlying

mechanisms in the disease process.
 Importance cont
 Clinical transplantation offers life saving
benefits in diseased organs or tissues
 Immunotherapeutic vaccines in the

control of many diseases (viral and

bacterial infections)
 Anthropological studies to determine

migration patterns
 Paternity identity and criminal

identification (employment of DNA and

HLA profiles)
 Historical Perspectives
First studies (1796) by Edward Jenner
 Showed body could respond to foreign

substances and generate the ability to defend

itself in subsequent infections.
 Exposure to cowpox led to resistance against

smallpox.
Scientist(s)
Edward Jenner (1749 – 1823)
Lowis Pasteur (1822)
Pasteur (1881 – 1885)
Ilya Metchnikoff (1845 – 1916)
Paul Ehrlich (1854 – 1915
Wright and Douglas (1903)
Von Behring (1854-1917) and
Kitasato (1870)
Robert Koch (1843-1910)
Buchner (1893)
Bordet (1895)
Contribution
Experimented with cowpox leading to small pox vaccine in
1796
Introduced vaccine with the first attenuated virus vaccine.
Germ theory led to production of attenuated vaccines
against anthrax, cholera and rabies.
Proposed “cellular theory of immunity” involving
“phagocytosis” of “wondering” cells (1884).
Believed “humoral immunity” involving antibodies and not
cells.
Demonstrated enhancement of phagocytosis by serum
opsonins indicating a linkage between humoral and cellular
immunity.
First described diphtheria antitoxin (antibody) – (1890).
Described delayed hypersensitivity reaction to tuberculin
(Koch’s phenomenon).
Described a heat labile serum factor (complement)
Demonstrated bacteriolysis of Cholera vibrio by antibody
and complement.
Ehrlich (1897) Proposed receptor theory of antibody synthesis and
first described neutrophils and eosinophils.

Landsteiner (1900) Discovered human ABO blood groups system

Von Pirquet (1874-1929) Described serum sickness

Fleming (1922) Identified lysozymes

Tiselius and Kobet (1938) Demonstrated that antibody activity resided in the
gamma globulin portion of serum proteins.

Paul Ehrlich (1892) Showed maternal transfer of species specific

immunity to infant through milk.

Calmette Guerrin (1920) Developed avirulent BCG vaccine.

Colonel Ogden Bruton Found agammaglobulinaema in a male child.

(1952)
Glick and Change (1956) Observed that antibody production in chickens
dependent on the presence of the Bursa of Fabricius
in birds
Henson (1739-1774) Described lymphocytes and were stained and
studied by Paul Ehrlich (1879).

Holmes et al (1966) Described phagocytic defect, chronic granulomatow

disease, in children.
Doherty and Zinkernagel Demonstrated that T cell recognition of antigen was
(1974) self –MHC restricted (1996).

Susumu Tonegawa (1976) Discovered that single immunoglobulin proteins

were encoded by separate rearranging genes (1987).

Kohler and Milstein (1975) Discovered the principle for production of

monoclonal antibodies

Porter (1950s and 1960s) Showed immunoglobulins were composed of two

heavy and two light chains covalently bonded (1950).

Max Theiler (1938) Contributed to development of yellow fever vaccine

BRANCHES OF IMMUNOLOGY
Basic Immunology
 Immunochemistry(Physiochemical

Properties)
 Cellular (Cells) and Molecular

(Molecules) Immunology
Clinical Immunology: Immunodeficiency;
Allergic and Hypersensitivity; Autoimmune
Diseases; Reproductive Immunology;
Immunohaematology; Tumour Immunology
and ClinicalTransplantation
Classification of Immune Responses
Body defence mechanisms
 Innate and adaptive immunity.

Humoral responses involve

 Soluble components including

immunoglobulins (antibodies) and

complement proteins
Cellular (cells) responses
 Lymphocytes, macrophages and

natural killer (NK) cells (principal

components)
 Outstanding Features
Adaptive immunity
 Recognition, narrow specificity and

memory.
Innate immunity
 Constituted by inborn defense

mechanisms
 Broad specificity, no memory
 Characteristic Features of Immunity
Innate Immunity Adaptive Immunity

Inborn pattern recognition receptors Randomly generated receptors

(PRRs) recognize pathogens recognize pathogens

PRRs possess a broad specificity for Receptors specific for particular

various Pathogen Associated epitopes : B cell receptor (BCR) and T
Molecular Patterns (PAMP) cell receptor (TCR)

Response immediate Response slow (3 - 5days) because of

immune system stimulation

No memory of prior exposure Memory of prior exposure exists

 Innate Mediators
Innate immune responses involve several
determinants (factors)
 Genetics, anatomical barriers, bacterial

antagonisms
 Pattern-recognition receptors (PRR)

 Nonspecific defense chemicals

 Inflammation and fever.

 Innate mediators cont
Phagocytic cells
 Neutrophils, monocytes and macrophages
 Basophils, mast cells and eosinophils release
inflammatory mediators.
Soluble factors
 Alternative pathway of complement activation
 Provides defense against gram-negative bacteria
 Interferons inhibit viral replication and activate
inflammatory cells.
 Genetic Determinants
Ir genes
 Determine susceptibility or resistance

and clinical expression of various

infections
HLA genetic markers
 Linked to susceptibility rather than

resistance to particular diseases

(autoimmune diseases)
 Genetic Factors in Malaria
Innate protection associated with
 Negative Duffy (a-b) blood groups (Plasmodium
vivax)
 Sickle cell trait A/S
 Plasmodium parasite infected erythrocytes
highly susceptible to toxic oxygen intermediates
or radicals.
 Intracellular development of P.falciparum in G-
6-PO4 dehydrogenase deficient erythrocytes
inhibited or retarded.
 Physical Barriers
Skin provides
 Intact outer horny layer mechanical
barrier against most pathogens and their
products
 Protective interface between internal
organs and the environment.
Dermis contains lymphocytes, mast cells
and tissue macrophages
 Epidermis equipped with
immunocompetent cells (Langerhans,
keratinocytes, epithelial cells, dendritic
cells and epidermal T lymphocytes)
 Physical Barriers Cont
Mucous, air-blood barriers and cilia lining the
respiratory tract
 Prevent adherence onto this portal entry by

infectious agents.
 High viscosity mucous and beating cilia (upper

and lower respiratory tracts)

 Trap and expel inhaled micro-organisms and

noxious agents through mucociliary system

mediated (mucociliary elevation)
 Mucous gels are inherently sticky and the layer

provides a barrier to parasite establishment

like intestinal worms.
 GIT barriers
Epithelium of the mouth and tongue
protected by antimicrobial peptides
 Stomach protected by pepsin digested

antimicrobial peptides and by low pH of

gastric juice
 Colon supports commensals prevented from

invasion of its lining by antimicrobial

peptides.
 Physical Barriers Cont
Human large intestine (colon) contains
enormous populations of commensal
bacteria with several benefits
 Synthesize vitamins

 Digest polysaccharides without human

enzymes
 Prime adaptive immunity.
 Physical Barriers To Infection

Barrier site (first line of defense) Activity

Skin sweat Flushing, organic acids

Skin and GI tract natural fauna

(commensals) Compete for niches

Peristalsis, low pH, bile acid,

GI tract flushing, antibacterial peptides

Lung tracheal cilia Mucociliary elevation

Nasopharynx, mucus and saliva, eye tears Flushing, lysozymes

 Recogniton
Innate immunity recognizes a few highly
conserved structures present in many different
microorganisms (pathogen – associated
molecular patterns, PAMPs) eg
 Lipolysaccharide (LPS) or endotoxin from
gram-negative bacteria cell wall
 Peptidoglycan, lipotechoic acids from gram-
positive cell wall, the sugar mannose (common
microbial glycolipids and glycoproteins)
 Bacterial DNA, N-formylmethionine found in
bacterial proteins
 Double stranded RNA from viruses and
glucans from fungal cell walls.
 Toll-like Receptors
 Pattern-recognition receptors (PRRs) recognize
common PAMPs of microorganisms.
Macrophages, dendritic cells and epithelial cells
express
 Transmembrane receptors that recognize
different types of PAMPs designated Toll-like
receptors (TLRs) eg
 TLR-2 that binds to peptidoglycan of gram-
positive bacteria (streptococci and
staphylococci);
 TLR-3 binds to ds RNA of viruses

 TLR-4 binds to LPS (endotoxin) of gram –

negative bacteria (Salmonella and E.coli).
 TLRs and Functions
Binding of gram-positive bacteria to TLR-2 and
gram-negative bacteria to TLR-4

 Enhances phagocytosis and fusion of the

phagosomes with lysosomes. TLR-5 binds
to flagellin of motile bacteria (Listeria).
 TLR-7 and TLR-8 bind to single stranded RNA
(ssRNA) genomes of various viruses (influenza,
measles and mumps).
 Opsonization and Phagocytosis
Opsonization involves opsonins
 Fibronectin (cold insoluble globulin)

 Specific IgG antibodies (Fc-fragment),

 C3b and iC3b, C3 products that facilitate

adherence of pathogens and membrane
perturbation.
 Phagocytosis process
Involve
 Formation of phagosome, fusion of phagosome
with lysosome and signal transduction leading
to stimulation of calcium intracellular influx.
 Activation of protein kinase C and
phospholipase occurs by diacylglycerol
releasing acid hydrolases and proteolytic
enzymes.
 Secondary granules contain lactoferrin,
gelatinase; complement receptors (CR1 and
CR3) and cytochrome B558 responsible for
intracellular killing machinery
Events in Phagocytosis involve (1) organism (bacterium) attaches to pseudopodia (long membrane evaginations) leading to (2) ingestion occurs forming
phagosome that (3) fuses with lysosome releasing lysosomal enzymes into phagosome and (4) digestion of ingested organism leading to (5) release of
products from the cell.
Source; http://www.whfreeman.com/COH1/phagocytosis.htm
 Antimicrobial Peptides
Antimicrobial peptides include defensins and
cathelicidins
 Protect against bacteria and other pathogens.

Defensins
 Secreted by epithelial cells (skin, GIT,
genitourinary tract and nasal passages and lungs)
or by recruited leukocytes (neutrophils).
 Punch lethal holes facilitated by their positive
charges in penetrating the bacterial membranes.
Cathelicidins
 Secreted by epithelial and neutrophils with
secondary alpha helix structures.
 Synergistically with defensins confer protection
against microbes.
 Microbicidal Mechanisms
Microbicidal mechanisms associated with
 Assembly of NADPH oxidase
 Upregulation of cytochrome B558 in

the activated neutrophils leading to

production of ROI (superoxides,
hydrogen peroxide)
 Formation of chlorinated derivatives

from hydrogen peroxide reaction with

chlorides generating hypochloric acid,
a microbicidal agent.
 Macrophage Derived Factors and Activities
Products Activity

Metabolites:
Reactive oxygen intermediates (ROI)
Reactive nitrogen intermediates (RNI)
Eicosanoids, prostaglandins leukotrienes
Platelet activating factor
Inflammation and intracellular killing
Inflammation and intracellular killing
Regulation of inflammation
Recruitment and activation of platelets.

Cytokines:
IL-1, TNF-α, IL-6 Inflammation
IFN-α Th1 activation
IL-10 Th1 suppression, Th2 activation
IL-12, IL-18 Activation of NK and T cells
TGF- β Inflammation, tissue repair
Adhesion molecules:
Fibronectin Opsonisation
Thrombospondin Adhesion, phagocytosis

Complement:
C3b, C4b and C2b Opsonisation

Enzymes:
Lysozyme Degrades bacterial cell walls
Collagenase, elactase Matrix catabolism
 Natural Killer (NK) Cells
 Large granular lymphocytes
 Posses CD16, CD56, CD 94, lectin-like
and Ig-like receptors.
 Provide early MHC independent
defence against intracellular infections
(herpex group viruses, Leishmania and
Listeria).
 Activated by IFN-α and IFN-β in
response to double stranded RNA
viruses.
 NK cont
 Produce IFN-γ that prime mφ to
secrete TNF and IL-12.
 Primed NK cells kill viral and
tumour cells by apoptosis and
recognize target cells that fail to
express class I MHC molecules on
their surfaces.
 Inflammation
 Represents generalized response to infection
or tissue damage
 Designed to localize invading

microorganisms and arrest the spread of

the infections
Characterized by 4 symptoms: Heat,
 Pain

 Redness

 Swelling
 Inflammation cont
Mast cells initiate inflammation mediated
by granules exocytosed when gram-
negative bacteria derived LPS bind
TLRs.
 Released histamine responsible for
redness and swelling associated with
inflammation.
 Tumour necrosis factor-alpha (TNF-α )
and chemotactic cytokines secreted by
stimulated mast cells.
 Inflammation cont
Reactive oxygen species and nitric oxide
produced by activated phagocytes
 Toxic to microorganisms and may also
lead to tissue injury.
 Macrophages and monocytes main source
of interleukin-1 (IL-1) causing fever.
 Arachidonic acid derivatives (leukotrienes
and prostaglandins) potent mediators of
inflammation
 Inflammatory response
Lyses of blood cells and release
 Bradykinin and prostaglandin cause the

pain associated with inflammation

 Lysosomal enzymes from damaged white

blood cells
 Serotonin from platelets

 Prostaglandins from damaged cell

membranes
 Acute phase proteins
Acute phase respones
 Leads to elevated production of protective

“acute phase” proteins by greater than 100

fold (C-reactive proteins, serum amyloid A
protein and α2 –macroglobulin).
 Acute phase proteins mediate restoration of
tissue integrity by restricting damage to the
injured site.
 Summary of Innate Determinants
 Genetics: Ir and HLA genes
 Anatomical (physical barriers): skin,
mucociliary elevation, cough reflex, wax
 Antagonism: commensal bacteria
 Soluble factors/chemicals: alternative
complement proteins, IFNs, cytokines,
lysozymes, lactoferrin, antimicrobial
peptides, acids
 Cells: phagocytes, granulocytes, NK cells
 Summary cont
Inflammation and fever
 PMNL recruitment and activation

 Bradykinins and prostaglandins

 Histamines

 Lysosomal enzymes

 Serotonin from platelets

 Reactive oxygen intermediates

 Cytokines (IL-1, TNF)

 Acute phase proteins

 Activation of Adaptive Immunity
Innate immunity may trigger adaptive immune
responses thru
 Antigen processing and presentation by

macrophages and dendritic cells

 Induction of costimulatory molecules (CD80/86)

by interaction with PAMPs and TLRs, CD28

binding and full T cell activation.
 PAMPs (LPS) bind to TLR on B cells enhancing

their response to antigen.

 Several adjuvants contain PAMPs thus

augmenting further adaptive response to

vaccines.
 Heat shock proteins (HSP)
 HSP linked to activation of adaptive
immunity.
 Family of substances produced when cells
and various infectious agents are exposed to
elevated temperatures.
 Immune System Components
Major histocompatibility complex (MHC) system
 Chromosome, gene, haplotype, and polymorphism
and super gene family molecules.
Humoral immunity consists of
 B-lymphocytes, bone marrow, stem cells, plasma
cells, immunoglobulins (Igs) – IgG, IgM, IgA, IgE
and IgD classes, class switching, antibody,
gammaglobulins and complement system proteins.
Cell mediated immunity comprises
 Lymphocytes(CD1,CD2, CD3, CD4, CD8),
monocytes-macrophages, NK cells and cytokines
 Concepts and Principles
Antigenicity and immunogenicity require
definition of
 Antigen, immunogen, hapten, carrier,
mitogen, alloantigen and an adjuvant.
Antigen-antibody reactions involve the
understanding of
 Immune complex (IC), epitope, agretope,
affinity, avidity, cross-reactivity,
precipitation reactions and agglutination
reactions.
 Antigens
Substance which induce and react with immune
responses (B or T or B+T)
 Proteins

 Glycoproteins and lipoproteins

 Synthetic peptides

 Alloantigens (transmembrane proteins)

 Nucleoproteins, hormones, drugs, metals

 Polysaccharides and glycolipids

 Pathogen (microbial) components and vaccines

 Microbial derived antigens
Entire cell (virus, bacteria and tumour
cells), parasite surface coat (protozoa) or
products (toxins).
Conserved proteins major antigens in infection
 Aldolase (target of human antibody
response against P. falciparum)
 Myosin (antigen in S. mansoni)
 Superoxide dismutase (M. leprae) and
cyclophilin (E. granulosus).
 Sperm antigens
Sperm surface antigenic determinants and the
seminal plasma constituents antigenic
 Sperm X-Y specific antigens induces immune
responses.
Fluid transport of spermatozoa (seminal plasma)
contains variety of potentially antigenic
components eg
 Decapitation factors, immunosuppressive
substances,
 Low molecular weight nitrogenous substances,
polyamines (spermine and spermidine),
 Carbohydrates (fructose, inositol) and
prostaglandins.
 Sperm Antigen cont
Lactoferrin
 Iron-binding protein

 Adsorbs onto the sperm and forms a major

component of sperm-coating antigen.

 Vaccine Ags
 Live attenuated viruses (Sabin for polio,
measles, small pox, hepatitis, yellow fever);
attenuated bacterial vaccines (BCG) or
 Inactivated microorganisms (Salk for polio,
typhoid, cholera, Pertussis)
 Subunit vaccines (influenza, meningococcal
and Pertussis antigens).
 Alloantigens
Transmembrane proteins eg
 Major histocompatibility complex (MHC)

antigens
 Blood group antigens.

 T cell independent antigens activate B

cells directly (polysaccharides) without T

cells help
 T cell dependent antigens activate B cells

with T cell help (mainly proteins).

 Superantigens
Microbial proteins produced by bacteria or
viruses capable of stimulating massive T
cell functions.
 Trigger diseases eg staphylococcal toxic
syndrome, staphylococcal food poisoning
and long-term effects including
 Autoimmune diseases (rheumatic arthritis,
multiple sclerosis, diabetes and rheumatic
fever), Kawasahi syndrome, atopic
dermatitis and periodontal disorders.
 Hapten and Carrier
Hapten: substance unable to elicit an immune
response and yet readily react with preformed
products of the induced response eg
 Drugs, cyclosporin, biotin, polysaccharides,
luteinizing-hormone-releasing factor (LHRF)
and dinitrophenyl (DNT).
Carrier: substance used in the modification of the
hapten renderng it immunogenic eg
 Staphylococcal enteroxin B; influenza virus
proteins; human serum albumin (HSA); bovine
serum albumin (BSA); fibrinogen and
concanavalin A (Con A).
 Factors Influencing Immunogenicity
High degree of foreignness
 Capacity of the body to discriminate

between self and non-self components.

 Either a productive immune response or

tolerance may be generated depending on

the degree of foreignness of the antigen.
 Molecular size and complexity
Epitopes or antigenic determinants
 High molecular weight substances greater than

6kDa (albumin, tetanus toxoid) highly

immunogenic
 Between 1-6 kDa (insulin, adrenocorticotropic

hormone) less immunogenic

 Less than 1 kDa (penicillin, aspirin,

progesterons) not immunogenic.

 Adjuvants
 Substances administered together with antigens
to improve their immunogenicity.
 Facilitate activation of accessory cells and

production of cytokines &provide signals for

stimulation of lymphocytes.
Most common inorganic adjuvants
 Aluminium hydroxide (alum); aluminium

phosphate; potassium ammonium sulphate and

calcium phosphate.
 Antigen Administration
Routes employed
 Intravenous; intradermal; intraperitoneal;
subcutaneous; submucosal and direct antigen
placement on/or in the mucosa.
Generally
 High immune responses induced when antigens
administered subcutaneously or intradermally
 Intravenous applications tend to activate
suppressor or inhibitory mechanisms
 Subcutaneous inoculation superior to
intramuscular antigen administration.
 Age determinant
 Immature/preterm and old age individuals
associated with diminished immune activity
 Poor and high responders associated with
immune response genes
 Summary Immunogenicity Characteristics
 Molecular size

 Degree of foreignness

 Chemical composition and texture

 Dose and route of antigen administration

 Genetic constitution and age of the

individual
 Haemopoitic System
Haemopoiesis occurs
 First in the yolk sac and foetal liver at 6 weeks, spleen

at 12 weeks and finally

 Bone marrow at 20 weeks of gestation as well as in

adult life.
Lymphoid lineage pathway gives rise to
 Thymus-derived (T) and bone marrow derived (B)

lymphocytes
Myeloid stem cells generate
 Mononuclear phagocytes (monocytes and

macrophages); polymorphonuclear lymphocytes;

mast cells; megakaryocytes and platelets.
Fig 3. Haemopoetic Cell Differentiation: Multipotent stem cells differentiate through either myeloid or lymphoid
pathways. Myeloid progenitor cells arte stimulated granulocyte-macrophage colony stimulating factor (GM-CSF) into
granulocyte-macrophage proginators, which differentiate into basophils, neutrophils or eosinophils, enhanced by IL-3,
G-CSF or IL-5 respectively. Monocyte- colony stimulating factor (M-CSF) stimulates granulocyte-macrophage
proginators into monocytes while EPO, TPO and IL-11 stimulate myeloid proginator cell differentiation into eventual
red blood cells and platelets. In the lymphoid pathway, B and T lymphocytes are derived from lymphoid progenitors
through differentiation of stem cells enhanced by IL-6 and IL-7 cytokines. Myeloid differentiation pattern is further
illustrated in Figs 3A and 3C.
 Growth Factors and Biological Activities
Growth Factors Source Biological Functions

PDGF Platelets, endothelial Promotes proliferation of

(Platelet Derived Growth cells, placenta connective tissue, glial and
Factor) smooth muscle cells

EGF Sub-maxillary gland, Promotes proliferation of

(Epidermal Growth Factor) Brunners gland mesenchymal, glial and
epithelial cells

TGF-α (Transforming Growth Common in transformed May be important for normal

Factor alpha) cells wound healing

FGF (Fibroblast Growth Wide range of cells Promotes proliferation of many

Factor) cells; inhibits some stem cells
and embryos

EPO (Erythropoietin) Kidney Promotes proliferation and

differentiation of erythrocytes

TGF-β (Transforming Growth Activated Th1 cells and Anti-inflammatory; promotes

Factor beta) natural killer (NK) cells wound healing; inhibits
macrophage and lymphocyte
proliferation.
 Lymphoid Cells
Distinguished by expression of
 Surface immunoglobulin (SmIg) on their cell

membranes.
Stimulation of B cells lead to the production of
 Various immunoglubulins mediating

humoral (antibody) immune responses.

Lymphocyte differentiation antigens
Most common cluster of human lymphocyte
lineage differentiation antigens (CD) exist for
both B cells and T cells.
 TABLE 6: Major B Cell And T Cell Differentiation Antigens

B Cell Types Expressing the Antigen T Cell Types Expressing the

Antigen Antigen Cluster Antigen
Cluster (CD) (CD)

CD 10 Committed B cell progenitors; CD 1 Thymocytes, dendritic cells

Pre-B cells
CD 19 All cells from early pre-B stage E-resetting T cells
to plasma stage (>90% of blood CD 2
B cells)
CD 20 Late pre-B to plasma stage cells T cell (pan T) associated with
CD 3 TCR
CD 21 All B cells in blood and Helper/Inducer T cells
lymphoid tissues CD 4
CD 22 Early pre-B cell stages to T cell (Pan T)
terminal plasma cell phase CD 5
CD 23 Small activated B cells in tissue T cell (Pan T): some B cells
CD 6
CD 24 B cell progenitors T cell (Pan T)
CD 7
CD 25 Activated T and B cells Cytotoxic/suppressor T cells
CD 8
CD 37 Late-B cell stage and early Early B cells, some T cells and
terminal plasma CD 10 granulocytes
CD 38 Germinal center B cells Activated T and B cells
CD 30
CD 39 Mentlezone B cells Thymocytes; active
CD 38 lymphocytes
CD 40 B cells; follicular dentritic cells Supp/indu/T cells; B cells
(FDC) CD 45R
 Organs of Immune System
Pluripotent stem cells in the bone marrow
 Differentiate into B lymphocytes and T

lymphocytes.
Lymphoid immune system consists of
 Primary lymphoid (10 ) organs and

 Secondary lymphoid (20) organs.)

 Primary Lymphoid Organs
Three human primary lymphoid organs
 Thymus, bone marrow and Peyer’s patches
Thymus arises from endoderm of the third
and fourth pharyngeal pouches
 A flat and bilobed organ above heart and

below the thyroid gland

 Largest relative size at birth and actual

largest size at puberty.

 Thymus cont
After puberty the thymus shrinks and T cell
production declines
 Children born without a thymus suffer from

DiGeorge syndrome.
Each lobule organized into
 Cortex (infiltrated with rapidly dividing

lymphocytes, thymocytes)
 Medulla (containing visible epithelial cells).

Both cortex and medulla contain stroma cells

 Composed of loosely packed thymic
epithelial cells, intergitating dendritic
cells and macrophages
 Thymus cont
Differentiation and maturation of T cells
mediated by
 Thymic epithelial cells and derived

 Hormonal factors (α -thymosin, β 4-

thymosin, thymopoietin, thymulin) and
IL-7.
A series of gene rearrangements select
antigenic diversity of T cell receptor
(TCR).
 Thymus cont
Thymocytes undergo thymic education
 Thymic cortical epithelial cells function in

positive selection process.

 T cells which bear TCR binding self-MHC

molecules are selected to survive and

proliferate (positive selection)
 Vast majority of remaining (95-99%) are

eliminated by programmed cell death

(apoptosis).
 Selection Process
Positive selection occurs when
 Double positive T cells bind cortical
epithelial cells expressing class I or II
MHC molecules plus self peptides with a
high affinity for survival signal.
Negative selection occurs when
 Double positive T cells bind macrophages
and dendritic cells expressing class I or
class II MHC molecules plus self peptides
with high affinity for apoptosis signal
 Bone Marrow
All cells of blood originate from
 Pluripotent stem cells in the bone marrow

(site of B lymphocyte maturation) involve

stroma cells and IL-7.
Negative selection thru apoptosis involve pre-
B cells
 If they do not form functional Ig molecules

 Possess Ig molecules that recognize and

bind to self antigens.

B cells that survive negative selection (about
10%) leave the bone marrow through
efferent blood vessels
 Peyer’s patches
 Located in the human wall of the
intestine.
PP exits with
 Primary lymphoid function and another

 With secondary lymphoid function

 Bursa of Fabricious
BF a round, sac like structure located above the
cloaca of birds
 Contain epithelial cells with lymphoid cells.

 Follicles divided into cortex and medulla

Cortex contain macrophages, lymphocytes and

plasma cells.
 BF serves as site of B cell maturation and

differentiation
 Secondary Lymphoid Organs
Secondary lymphoid organs
 Sites where lymphocytes encounter

antigens
 Interact with other cells and enlarge in

response to antigenic stimulation.

 Poorly developed in germ-free animals.

Lymphoid organs morphologically divided

into: cortex, paracortex and medulla
 Second Lymph organ cont
 Cortex (outermost layer full of B lymphocytes
and macrophages arranged in primary follicles)
 Paracortex (beneath the cortex, populated with

T lymphocytes and dendritic cells)

 Medulla (innermost with sparsely populated

lymphocytes and plasma cells).

Lymphocyte circulation involve
 Naïve T cells that bind to endothelium of
venules in the paracortex
 T cells leave the bloodstream via
conventional blood vessels and carried to
the lymph nodes via tissue fluid (plasma).
A schematic presentation of secondary lymphoid
organ is provided in Fig. 8.

Fig. 8 Structure of a Secondary Lymphoid Organ: Refer to text for the details. Source: http://www-immuno.path.com.oc.uk
 Lymph Node
Lymph nodes
 Encapsulated and packed with lymphocytes,

macrophages and dendritic cells,

 Filter antigens from the lymph,

 Consist of cortex, paracortex and medulla.

Cortex (outer most layer) contains

 Mostly B lymphocytes, follicular dendritic cells

and macrophages all arranged in clusters

(designated primary follicles
 Antigen Stimulation
Following antigen stimulation the follicles become
 Secondary follicles consisting of

 Concentric rings (germinal centres) of densely

packed lymphocytes, macrophages and dendritic
cells.
Germinal centres
 Sites of intense B cell activation and
differentiation into plasma cells and memory cells.
Paracortex (just beneath the cortex) designated T
dependent region
 Populated with T lymphocytes and interdigitating
dendritic cells important in T cell activation.
 Medulla
Medulla (inner most regions)
 Sparsely populated by plasma cells, activated

Th and Tc cells and high concentration of Igs.

 Lymphocytes enter the lymph node between

specialized capillary endothelial cells in the

postcapillary venules designated high
endothelial venules (HEVS)
 Lymphatic vessels
Lymphatic vessels responsible for flow of
lymph within the lymphoid system.
 Fuid from the tissues flows from the

intercellular tissue spaces into lymphatic

capillaries and then into lymphatic vessels.
 Lymph flows through regional lymph nodes

and eventually enters the circulatory

system
 Other Lymphoid organs
Less organized aggregates of lymphoid
organs include
 Tonsils, appendix, Peyer’s patches in the

lining of small intestine (gut associated

lymphoid tissue or GALT)
 Lymphoid tissue beneath the mucous

membranes of the bronchi (bronchial-

associated lymphoid tissue or BALT)
 Other Lymph organ cont
 Aggregates of lymphatic tissue throughout
the mucous membranes (mucosal
associated lymphoid tissue or MALT) and
 Beneath the skin (skin associated lymphoid
tissue or SALT).
Pathogens and other antigens enter tissues,
transported by tissue fluid into lymphatic vessels.
 Regional lymph nodes
In the regional lymph nodes
 Microbes and antigens in the lymph

encounter changing populations of B-

lymphocytes
 Filtered out and phagocytosed by fixed

macrophages and dendritic cells.

 Antigen Uptake
When microorganisms and other antigens enter
blood
 Transported by blood vessels to the spleen

where they encounter changing population of B

lymphocytes
Germinal centers
 Secondary follicles found in tissues containing

activated cells following continuous exposure to

antigen.
 Sites of antigen driven activation of B cells in

secondary lymphoid organs

 Leads to the development of active B cells,

plasma cells and macrophages.

 B Cell Priming
Antigen specific B cells
 Activated in the T cell zones of secondary

lympoid tissue and

 Migrate to B cell zones to form germinal

centers within the reticulum of follicular

dentritic cells (FDCs).
 Primary follicles consist of a

 Network of FDCs in which no antigen-

driven process is occurring.

 Lymphocyte Priming
FDCs
 Depot of native unprocessed antigen, in an
immune complex form
 Hold Ag in a nondegraded form for long
periods.
Follicular B cells can
 Take up antigen from FDCs and process and
present in a form that activates T cells.
 T cell – B cell collaboration essential in the
development of germinal centers
 MHC System Genetics
9 MHC genes and include
 HLA-A, HLA-B, HLA-C, HLA-DPA 1, HLA-
DPB1, HLA-DQA1, HLA-DQB1, HLA-DRA
and HLA-DRBI.
MHC region divided into three regions
 Class I (HLA-A,B and C)

 Class II (HLA-DP, DQ and DR) and

 Class III genes encode complement components

(C2, C4 and Factor B), cytokines (TNF-α)
 MHC genes display high levels of allelic
diversity (polymorphism).
 Class 1 MHC Molecules
Class I MHC molecules consist one long heavy
chain of 346 amino acids and a short one of 99
amino acids.
Heavy chain consists of five regions
 A transmembrane domain and a cytotoplasmic

domain (with the C terminal).

 Three extracellular domains including N terminal,

 Protein molecule beta-2 microglobulin (β -M)

2
non-covalently attached to the heavy chain.
 Class I polymorphism
Class I MHC region encodes HLA-A, B
and C molecules.
 HLA-A, -B and -C heavy chain
genes are highly polymorphic with
known 60, 138, and 40 alleles,
respectively.
 Class II MHC Molecules
Human class II molecules are designated HLA-D and
class II MHC region encodes
 Molecules HLA-DP (α,β); HLA-DNA; HLA-DOB;
HLA-DQ (α,β) and HLA-DR (α,β).
Other encoded molecules are
 Transporter proteins (TAP 1 and 2) involved in antigen
presentation;
 Low molecular weight polypeptides (LMPs and 7) and
HLA-DM gene products.
MHC Class II molecules are found only on
 Professional antigen presenting cells (APCs) including
macrophages/monocytes, dendritic cells, activated T
cells andB cells.
 Class III MHC Molecules
Class III MHC region
 Located in the central part of the MHC system
and
 Contains genes which code for complement
components
 Factor B (BF), C2, C4b and C4a]; tumour necrosis
factor (TNF); lymphotoxin (LT); heat shock
proteins (HSP 70-1 and HSP 70-2).
 Proteins produced also include 21 –
hydroxylase enzyme (21-Ohase), deficient in
patients with congenital adrenal hyperplasia.
 MHC System Components

Class MHC Loci Tissue distribution of MHC gene

products

I HLA-A,B,C All nucleated cells, lymphoid, epithelial and

mesenchymal tissues

II HLA-DP, HLA-DNA, Accessory antigen presenting cells like

HLA-DOB, HLA-DQ monocyte-macrophage lineage cells, B-cells
HLA-DR and activated T-cells

III Central MHC C2,BF, C4b, C4a, and 21-hydroxylase

region enzyme (21- Phase), tumour necrosis factor
(TNF), lymphotoxin and heat shock
proteins
 MHC Region Characteristics
MHC Region posses characteristic
features
 Multiplicity, high polymorphism and

extensive heterozygosity.
High polymorphism
 Genes in the MHC loci exist in different

or alternative multiple forms designated

alleles.
 HLA Specificities (Alleles)
A B C D DR DQ DP
A1 B5 CW1 DW1 DR1 DQW1 DPW1
A2 B7 CW2 DW2 DR2 DQW2 DPW2
A3 B8 CW3 DW3 DR3 DQW3 DPW3
A9 B12 CW4 DW4 DR4 DQW4 DPW3
A10 B13 CW5 DW5 DR5 DQW5(W1) DPW5
A11 B14 CW6 DW6 DRW6 DQW6(W1) DPW6
AW19 B15 CW7 DW7 DR7 DQW7(W3)
A23(9) B16 CW8 DW8 DRW8 DQW8(W3)
A24(9) B17 CW9(W3) DW9 DR9 DQW9(W3)
A25(10) B18 CW10(W3 DW10 DRW10
A26(10) B21 ) DW11(W7) DRW11(W7)
A28 BW22 CW11 DW12 DRW12(5)
A29(W19) B27 DW13 DRW13(W6)
A30(W19) B35 DW14 DRW14(W6)
A31(W19) B37 DW15 DRW15(2)
AW34(10) B38(16) DW16 DRW16(2)
AW36 B39(16) DW17(W7) DRW17(3)
AW43 B40 DW18(W6) DRW18(3)
BW41 DW19(W6) DRW19(3)
BW42 DRW52
DR21

DRW53
NB. The alleles that have been reported and provisionally given a number pending verification and
official recognition are referred to by the letter “W” before the number such as Aw33(w19).
 Inheritance of Haplotypes
MHC genes are co-dominant and the alleles equally
expressed.
Given a simple Mendelian pattern of inheritance,
 Distribution of MHC haplotypes in a family: 25%

identical; 25% non-identical and 50% partially –

identical.
Linkage disequilibrium
 Certain gene combinations occur more frequently

than is expected given random Mendelian

combinations and permutations
 HLA-B8 and HLA-DR3 alleles more closely

linked (97%)
Alleles in Linkage Dysequilibrium
A1, B8 AW30 B13 DW1, BW35
A1, BW17 AW33, BW14 DW, BW7
A2, BW12 CW1, B5 DW4, BW15
A2, BW16 CW1, BW16
A11, B5 CW3, BW15
A29, B12 CW4, BW35
CW5, B12
 Importance of MHC Antigens
MHC antigens-a group of cell membrane
components
 Expressed on cells by certain individuals
in a population referred to as
alloantigens.
Clinical transplantation
 Matching donor HLA antigens with
those of recipient to avoid induction of
graft rejection reactions
 Patenity identity
HLA gene products of a child compared with
 Putative candidates to ascertain the
biological father in a given ethnic or
racial group.
 Match between the HLA antigens of

the child and candidates will establish

patenity.
 Anthropological studies
HLA closely linked and their distribution in the
population markedly restricted.
HLA polymorphism shows
 Sub-Saharan African populations
genetically most diverse (founding
human race probably of the African
descent).
Migration patterns of peoples in populations
determined through HLA characterization.
 Liberians appear more closely related to
a distinct Malawian population than to
their West African neighbours.
 Forensic use of HLA typing
 DNA probes used in resolving inconclusive
blood group typing or finger print results in
criminal investigations.
HLA and disease association
Theories include
 Linkage disequilibrium,

 Antigenic mimicry

 Role of various infectious agents

 Strong HLA disease association exists between
M.tuberculosis and HLA-Bw 15, M. leprae infections with
HLA-DR3.