IMMUNOLOGY AND SEROLOGY LECTURE

Immunology
Study of the components and processes involved in the combat and disposal of foreign material
Began as a branch of Microbiology
HISTORICAL BACKGROUND
Scientist(s)
Thucydides
Lady Montagu (British Ambassador)
Edward Jenner
Neisser
Louis Pasteur (Founder of Immunology)
Elie Metchnikoff
Klebs and Loeffler
Roux and Yersin
Emil von Behring and Shibasaburo Kitasato
Almroth Wright and S.R. Douglas
Paul Ehrlich
Charles Richet and Paul Porter
Clemens von Pirquet and Bela Schick
Karl Prausnitz and Heinz Kustner
Riley and West
Karl Landsteiner
Haurowitz
Niels Jerne and McFarlane Burnet
Dausset and Rapaport
Claman
Milstein and Kohler

Contribution(s)
Bubonic plague those who had the disease
could nurse patients and would not contract it
again
Introduced variolation
*Variolation introduction of materials from
lesions to induce resistance
Vaccination (from vacca=cow)
Isolated first human pathogen, Gonococcus
Chicken cholera vaccine
Anthrax vaccine
Rabies vaccine
Cellular Phagocytic Theory of Immunity or Cellmediated Immunity
Isolated diphtheria bacilli
Produced first defined antigen (diphtheria toxin)
Humoral Immunity
Immune response is both cell-mediated and
humoral
Opsonins
Standardization of diphtheria toxin and antitoxin
preparation
Phenomenon of prophylaxis
Anaphylaxis is both beneficial and damaging
Serum sickness
Allergies can be transmitted
Histamine is present in mast cells
ABO Blood group system
Template Theory of Antibody formation
Clonal Selection Theory
HLA (Human Leukocyte Antigen)
T-cells and B-cells work together in an immune
response
Hybridoma for the production of monoclonal
antibodies

IMMUNOLOGIC METHODS
Immunologic Method(s)
Complement Fixation

Discoverer(s)
Bordet and Gengou

Passive Hemagglutination
Boyden
Rosette Formation
Biozzi and Zaalberg
Plaque Formation by Immunocompetent Cells in Jerne, Henry and Nordin
Agar Gel
Use of Abs or Ags Labelled by Fluorescent
Coons
Compound
Use of Abs or Ags Labelled by Enzymes
Avrameas and Uriel
Use of Abs or Ags Labelled by Radioactive
Yalow and Berson
Elements
Precipitation in Gelled Media
Ring test (disk test)
Bechold
Disk test for diagnosis of anthrax
Ascoli
Simple diffusion method to quantitate antigenOudin
antibody complexes
Double diffusion method useful for qualitative
Ouchterlony and Elek
comparisons of antigens and antibodies
Immunoelectrophoresis Analysis in Gels
Grabar and Williams
Quantitative Modification of
Ressler and Laurel
Immunoelectrophoresis
LYMPHOID ORGANS
Lymphoid Organ(s)

Function(s)
Primary
Bone marrow
Site of B-cell maturation
Center for antigen-independent lymphopoiesis
Thymus
Produces T-cells
Secondary
Spleen
Has white and red pulp, removes old and
damaged cells
Filters antigens found in blood
Lymph nodes
Filter antigens found in lymph (tissue fluid)
Tonsils
Filter and trap antigens entering via the
nasopharyngeal or alimentary tract
Peyers Patches, Appendix, MALT (Mucosal
Filters antigens found in the gastrointestinal tract
Associated Lymphoid Tissue)
Immunity
Sum of all mechanisms involved in resisting infections
Has 2 types
o Natural/Innate/Non Specific Immunity doesnt require exposure to develop antibodies
o Acquired/ Specific Immunity requires exposure to develop antibodies

NATURAL/INNATE/NON-SPECIFIC IMMUNITY
External Defense System
Skin

Internal Defense System

Cellular factors (phagocytosis)

Mucous membrane

Humoral factors
Cytotoxic lymphocytes
Inflammation

Component(s)
Physical/Mechanical Barrier

Constituent(s)
Intact skin (keratin = inhibition of microbial
growth)
Viscous Mucus (cilia and mucous secretions)
Shedding of cells
Body fluids
Tears and saliva (lysozyme)
Urine
Sweat and fatty acids (lactic acid)
Bile salts and gastric acid
Vomiting and Diarrhea
pH (lactic acid in genital tract)
Body temperature
Oxygen tension (High OT = no anaerobes)
Hormonal balance
Age

Physiologic/Biochemical Factors

Phagocytosis, Lymphocytes, Inflammation,

Humoral Factors
Endocytosis entry of particles or materials into the cell
Phagocytosis cell-eating
Pinocytosis cell-drinking
PHAGOCYTOSIS
Ingestion of a particulate matter (e.g bacteria) to destroy it
Carried out by neutrophils, monocytes, macrophages
Factors Enhancing Phagocytosis
Integrins attaches the antigen to the phagocyte
Opsonins makes the antigen more susceptible for phagocytosis
Phagocytic Cells
1. Neutrophils
PMN with a nucleus of 2-5 lobes
Primary granules (azurophilic granules)
o Myeloperoxidase
o Elastase
o Proteinase 3
o Lysozyme
o Cathepsin G
o Defensins
Secondary granules

o Collagenase
o Lactoferrin
o Lysozyme
o Reduced NADPH (Nicotinamide Adenine Dinucleotide Phosphate) oxidase
Tertiary granules
o Gelatinase
o Plasminogen activator

2. Monocytes
Largest cell in peripheral blood having a horseshoe-shaped nucleus with a dull grayishblue cytoplasm and ground-glass appearance
Granules contain:
o Peroxidase
o Acid phosphatase
o Arylsulfatase
o Lipase
o Lysozyme
o -glucoronidase
3. Macrophages
Arise from monocytes
Functions mainly in phagocytosis and antigen presentation
Tissue Macrophage
Kupffer cells
Alveolar macrophage
Mesangial cells
Microglial cells
Synovial A cells
Histiocytes

Location
Liver
Lungs
Kidney
Brain
Joints
Connective tissues

Stages of Phagocytosis
1. Initiation
formation of cell surface receptors (CR3, Laminin, FMLP or Formyl Methionyl Leucyl
Phenylalanine) important for the attachment of antigens
2. Chemotaxis
Movement of phagocyte towards site of infection
Diapedesis movement of phagocyte into tissues
Mediated by chemotaxins substances that attract or help the phagocyte locate the site
of infection
3. Engulfment
Attachment of phagocyte to the antigen
Formation of phagocytic vacuole or phagosome
Respiratory burst occurs
4. Digestion or Cytopepsis

 Formation of phagolysosome (phagosome + lysosome)

Phagocytes granules act on the antigen
5. Excretion
Disposal of undigested materials via exocytosis
INFLAMMATION
Overall reaction of the body to injury or infection
Involves both cell-mediated and humoral immunity
Cardinal Sign(s)

Cause
Lactic acid production due to aerobic
glycolysis
Heat (calor)
Interleukin-1 secreted by monocytes
Redness (rubor)
Hyperemia (increased blood flow)
Swelling (tumor)
Leakage of plasma to tissues
*When these signs arent addressed, loss of function (functio laesa) occurs.
Pain (dolor)

Stages of Inflammation
1. Vascular response
Histamine (from mast cells) dilates blood vessels causing increased blood flow (heat and
redness occurs)
Blood vessels contract increasing permeability resulting to leakage of plasma to tissues
(swelling and pain take place)
2. Cellular response
Through the action of chemotaxins, neutrophils migrate to site of inflammation which
will be followed by monocytes then macrophages.
3. Cellular proliferation and repair
Fibrinogen initiates clotting to prevent further antigenic invasion.
Fibroblasts appear to replace dead cells.
HUMORAL FACTORS
1. Antibody and Complement System
Ab-activated complement lyses LPS (lipopolysaccharide) layer of bacterial cell wall
Lysozyme enters and destroys the mucopeptide layer
MAC (membrane attack complex) produces pores resulting to death of the bacterial cell
o Perforins substances that produce pores or holes on the cell membrane of
bacterial cell wall
2. Non antibody Humoral Factors
Chemotaxin 5a attracts phagocytes
Properdin activates alternative pathway of complement
IFN or Interferon proteins from virally-infected cells that protect neighboring cells
o Type I IFN- (macrophages) and IFN- (fibroblasts)

o Type II or immune IFN (T-cells)

-Lysine from ruptured platelets, directed to Gram (+) bacteria
Lactoferrin (milk) and Transferrin (serum) deprives bacteria of iron
Lactoperoxidase (milk and saliva) catalyst in the breakdown of hydrogen peroxide
(H2O2) into water (H2O) and oxygen (O2)
Lysozyme (serum, tears, saliva, nasal secretions, etc.) hydrolyzes mucopeptide layer of
bacterial cell wall

ACQUIRED/SPECIFIC IMMUNITY
Specific Immunity may be:
Cell-mediated
Involves T-cells
Achieved through direct cell-cell contact
Defense against viral and fungal infections,
and graft rejection

Humoral
Involves B-cells
Composed of antibodies
Primary defense against bacterial infection

CHARACTERISTICS
1. Ability to distinguish self from non-self
2. Specificity (particular antibodies act on particular antigens)
3. Immunologic memory (ability to remember the antigen to which antibodies react)
Ways on How to Acquire Specific Immunity
Mode of Acquisition
Active
Passive
Active
Passive

Example(s)
Naturally Acquired
Convalescence from an infection
Transplacental passage of antibodies from mother
to fetus
Artificially Acquired
Immunization or vaccination
Injection of immune globulins

Active Immunization vs. Passive Immunization

Active immunization administration of the antigen itself (attenuated, killed or extracted
components) to induce an immune response
Passive immunization administration of immune globulins (antibodies); has 3 types of
preparations antitoxin, immune globulin, & specific immune globulin
Routes of Administration
1. Intramuscular and subcutaneous
2. Intradermal or intracutaneous

3. Oral
4. Intranasal

Vaccines
May be composed of killed antigen (with the use of chemicals or heat), live attenuated antigens,
or antigenic materials (cell wall, capsule, pili, etc.)
Type(s) of Vaccine

Bacterial

Representative Vaccines
Diphtheria toxoid

Tetanus toxoid
Pertussis (whooping cough)
Bacille-Calmette-Guerin (BCG)
Typhoid vaccine
Pneumovax
Haemophilus influenza vaccine
N. meningitides vaccine

Rubella (German measles) vaccine

Influenza virus vaccine
Measles and mumps vaccine
Poliomyelitis vaccine
Viral

Rabies vaccine
Hepatitis B vaccine
Typhus vaccine
Ricketsial

Rocky Mountain spotted fever

vaccine

Preparation
Precipitation with potassium
alum (tawas) or adsorbed
onto aluminum hydroxide
Inactivated C. tetani toxin
Thimerosal or heat-killed
B.pertussis
Live attenuated M.bovis
Heat or acetone-killed
S.typhi
Strains of S.pneumoniae
Polysaccharide capsule of H.
influenzae
Capsular carbohydrates of
meningococci
Live attenuated virus in
tissue culture
Whole virus or disrupted
virus products inactivated by
formalin or UV light.
Live attenuated virus grown
in chick embryo
Salk vaccine (inactivated
polio vaccine)
Sabin vaccine (live
attenuated oral polio
vaccine)
Inactivation by propiolactone or tributyl
phosphate
Purified,inactivated, alumadsorbed HbsAg
Formalin-killed Ricketssia
prowazeki
Formalin-killed R.rickettsi

Passive Immunization
Preparation

Representative vaccines

Antitoxin

Botulism antitoxin, Diphtheria antitoxin, Tetanus

antitoxin, Antivenom
Human-derived gamma globulin
Hepatitis B immune globulin; Human-derived
rabies, varicella-zoster, tetanus immune globulin

Immune Globulins
Specific Immune Globulins

ANTIGENS/IMMUNOGENS
Immunogens vs. Antigens
Immunogens macromolecules capable of triggering an immune response
Antigens foreign materials which cannot induce an immune response.
All immunogens are antigens but not all antigens are immunogens.
Factors Affecting Immune Response
1. Age
Neonates ( - immunity not well developed)
Old individuals ( - immunity deteriorating)
2. Overall Health
Malnutrition, fatigue, stress ( resistance to infection)
3. Dose of Inoculation
Higher dose = greater immune response
*Dosage of immunogen needed differs with route of inoculation.
4. Genetic Predisposition
MHC
T-cell and B-cell receptors
Characteristics of Immunogens
1. Macromolecular size
Greater mol. wt. = More potent immunogen
At least 10,000 daltons recognizable immunogen
Best immunogens mol. wt. of >100,000 daltons
2. Chemical Composition and Molecular Complexity
Proteins & polysaccharides best immunogens
Proteins
Made of amino acids
Has primary structure (sequence of amino acids), secondary structure (orientation of
amino acids w/in the chain), tertiary structure (spatial or 3D orientation of the
molecule) and quaternary structure (2 or more chains associate to make a single
polymeric unit)

Carbohydrates
Less immunogenic than proteins
Glycolipids and glycoproteins immunogenic carbohydrates
Blood group Ags are carbohydrate complexes (A,B,H antigens glycolipids; Rh
and Lewis antigens glycoproteins)
3. Foreignness
Acquired when lymphocytes mature in primary lymphoid organs
more distant taxonomically the source of immunogen from the host = better
immunogen
autoantibodies antibodies against self cells
ability to be processed and presented with MHC molecules
Epitopes
antigenic determinant or portion of Ag that binds with an Ab
molecular shapes or configurations recognized by B and T cells
may be sequential, linear, of repeating copies or different specificities
conformational epitope - the folding of one chain or multiple chains, bringing certain amino
acids from different segments of a linear sequence or sequences into close proximity with each
other
T cells recognize an epitope only as a part of a complex formed with MHC proteins on the
surface of an antigen-presenting cell
Haptens
Non-immunogenic materials that when combined with a carrier (e.g protein) may induce an
immune response
Incomplete or partial immunogen
Agglutination and precipitation cannot occur
Examples
o Catechols (in poison ivy) results to contact dermatitis
o Drug-protein conjugate (like penicillin) may cause life-threatening allergic reactions
Antigen Host Relationship
1. Autoantigen
Belongs to the host
Doesnt induce immune response under normal condition
2. Alloantigen
From other members of same species
Induce immune response
Tissue transplantation and blood donation
3. Heteroantigen
From other species

Heterophile antigens Ags from other species but are identical or closely related to another
species

ANTIBODIES/IMMUNOGLOBULINS
Antibodies
Humoral branch of immune response
Neutralize toxic substances, facilitate phagocytosis and microbial killing, and combine with
antigens on their surface resulting to lysis of these cells extravascularly (mononuclear-phagocyte
system) or intravascularly (through complement system)
Antibody Structure

Consist of 2 heavy chains and 2 light chains

Light Chain
23,000 daltons composed of 200 amino acids
Has kappa () and lambda () with a ratio of 2:1 respectively
Heavy Chain
50,000-70,000 daltons composed of about 400 amino acids
Has five distinct isotypes
Gamma (contains gamma chain)
Mu (contains mu chain)
Alpha (contains alpha chain)
Delta (contains delta chain)
Epsilon (contains epsilon chain)

Disulfide bonds hold the four polypeptide chains

Interchain Heavy to Heavy, Light to Light, and Heavy to Light
Intrachain disulfide bonds holding the amino acids in one polypeptide chain

Regions

Variable region variety of amino acids in the amino terminal portion of the polypeptide chain
Constant region unvarying sequence of amino acids in the carboxy terminal portion of the
polypeptide chain

Domains

Series of globular homology regions

Immunoglobulin

Constant

Variable

IgG
IgM
IgA
IgD
IgE

3
4
3
3
4

1
1
1
1
1