Good Morning

Under the guidance of : Dr. Sneha Mantri Maam Dr. Chandan Rathod Sir

INTRODUCTION
The term immunity refers to the resistance exhibited by the host towards injury caused by microorganisms & their products. Protection against infectious disease is only one of the consequences of immune response which in its entirety is concerned with the reaction of the body against any foreign antigen.

Immunology
The Study Of Immune System Latin Word immunisexempt Earliest Written Reference was Thucydides 430 BC Pasteur Was First To Successfully Apply Vaccination

Humoral or Cellular Immunity

Pasteur did not know how vaccination worked Behring and Kitasato (1890) proposed serum was responsible for Immunity Elvin Kabat (1930), gamma-globulin, antibody Antibodies were present in body fluidsHumor Therefore: Humoral Immunity

Innate Immunity

Innate Immunity made up of 4 forms Anatomical, physiological, phagocytic and inflammatory Anatomical: skin, epidermis (densely packed dead cells) Flow of mucus prevents bacterial entry by washing them away Normal flora colonize epithelial cells of mucosal surfaces, pathogens compete with them for attachment sites.

Cell Mediated Immunity

In 1883 Ellie Metchnikoff showed that cells responsible for Immune state Phagocytes more active in Immune animals He hypothesized that cells responsible for Immunity, not serum components Controversy developed but Humoral School prevailed till 1940 Merrill Chase experimented (1940) with tuberculosis infected animals, immunity through White Blood Cell transfers

Immune System
It includes all parts of the body that help in the recognition and destruction of foreign materials. White blood cells, phagocytes and lymphocytes, bone marrow, lymph nodes, tonsils, thymus, and your spleen are all part of the immune system.

Components of Immune System

Classification

Innate (native) immunity Non specific: species racial individual Specific : species racial individual

Acquired (adaptive) immunity Active : natural artificial Passive : natural artificial

Innate immunity
Also known as Native Immunity Resistance to infections which an individual possesses by virtue of his genetic & constitutional makeup Not affected by prior contact with microorganisms or immunisation.

Components of Innate Immune system

First line
1) Mechanical barriers 2) Chemical & biochemical inhibitors 3) Normal flora

Second line
A- cells 1- Natural killer 2- Phagocytes B- Soluble factors C- Inflammatory barriers

Mechanical barriers
- Intact skin - Mucous coat - Mucous secretion - Blinking reflex and tears - The hair at the nares - Coughing and sneezing reflex

Chemical & biochemical

inhibitors
- Sweat and sebaceous secretion - Hydrolytic enzymes in saliva

- HCl of the stomach

- Proteolytic enzyme in small intestine

- Lysozyme in tears
- Acidic pH in the adult vagina

Normal bacterial flora

- Competition for essential nutrients

- Production of inhibitory substances

Cells
Natural killer (NK) Definition: Source :
Large granular lymphocytes Innate cytotoxic lymphocytes Bone marrow precursors 10% or 15% of lymphocytes in peripheral blood 1% or 2% of lymphocytes in spleen

Location : Function

Tumor cells : Cytotoxic for Viral infected cells Bacterial, fungal, parasitic infection Responsible for antibodydependent cell mediated cytotoxicity

Phagocytes
Specialized cells for capture, Ingestion and destruction of invading microorganisms * Polymorphonuclear leucocytes, mainly neutrophils: granulocytes circulate in blood

* Mononuclear cells (macrophages) - Monocytes in blood

- Histiocytes in connective tissues
- Fixed reticuloendothelial spleen, lymph nodes, bone marrow

cells in liver,

Phagocytosis

Phagocytes & their Relatives

Neutrophils
60% of WBCs Petrol tissues as they squeeze out of the capillaries. Large numbers are released during infections Short lived die after digesting bacteria Dead neutrophils make up a large proportion of pus.

Macrophages

Larger than neutrophils. Found in the organs, not the blood. Made in bone marrow as monocytes, called macrophages once they reach organs. Long lived Initiate immune responses as they display antigens from the pathogens to the lymphocytes.

Lymphocytes

Produce antibodies B-cells mature in bone marrow then concentrate in lymph nodes and spleen T-cells mature in thymus B and T cells mature then circulate in the blood and lymph Circulation ensures they come into contact with pathogens and each other

B- Lymphocytes
There are 10 million different Blymphocytes, each of which make a different antibody. The huge variety is caused by genes coding for abs changing slightly during development. There are a small group of clones of each type of B-lymphocyte

B- Lymphocytes
At the clone stage antibodies do not leave the B-cells. The abs are embedded in the plasma membrane of the cell and are called antibody receptors. When the receptors in the membrane recognise an antigen on the surface of the pathogen the B-cell divides rapidly. The antigens are presented to the B-cells by macrophages

B- Lymphocytes

Some activated B cells PLASMA CELLS these produce lots of antibodies

The antibodies travel to the blood, lymph, lining of gut and lungs. The number of plasma cells goes down after a few weeks Antibodies stay in the blood longer but eventually their

numbers go down too.

T- Lymphocytes

Mature T-cells have T cell receptors which have a very similar structure to antibodies and are specific to 1 antigen. They are activated when the receptor comes into contact with the Ag with another host cell (e.g. on a macrophage membrane or an invaded body cell)

T - Lymphocytes
After activation the cell divides to form: T-helper cells secrete CYTOKINES help B cells divide stimulate macrophages Cytotoxic T cells (killer T cells) Kill body cells displaying antigen Memory T cells remain in body

Soluble factors
1- Acute phase protein :Plasma protein, CRP=C reactive protein, Fibrin. 2- Complement : proteins in serum, body fluids 2- Interferons : Proteins against viral infections 3- Properdin : Complement activation 4- Beta lysine: Antibacterial protein from Platelets 5- Lactoferrrin,Transferrin :Iron binding protein 6- Lactoperoxidase : Saliva & Milk 7- Lysozyme : Hydrolyze cell wall

Interferons
Proteins usually produced by virally infected cells Types of interferons: 1- Alpha interferon Secreted by
Induced by
Polynucleotide Macrophages Viruses or

2- Beta interferon

Secreted by Fibroblasts, Viruses

3- Gamma interferon T- lymphocytes, Specific antigens

Protective action of interferons

1) Activate T-cells
2) Activate macrophages 3) Activate NK

Inflammatory barriers
* Tissue damage by a wound or by invading pathogen * Inflammatory response: Tissue damage Release of chemical mediators from Leukocytes (Histamine, fibrin, kinins, cytokines) Invading microbe Redness of tissue Tissue temperature Capillary

Vasodilatation of capillaries permeability

Influx of fluids Influx of phagocytes into tissues

Inflammation

Acquired Immunity
Resistance that an individual acquires during life is known as acquired immunity Distinct from inborn innate immunity Two types- active & passive

Active & Passive Immunity

ACTIVE IMMUNITY
Produced actively by hosts immune system Induced by infection or by immunogens Durable effective protection Immunity effective only after lag period Immunological memory present Booster effect on subsequent dose negative phase may occur Not applicable in immunodeficient

PASSIVE IMMUNITY
Received passively, no active host participation Readymade antibody transferred Transient, less effective Immediate immunity No memory Subsequent dose less effective No negative phase Applicable in immunodeficient

Active & Passive Immunity

Vaccination
A preparation containing antigenic material: Whole live microorganism Dead microorganism Attenuated (harmless) microorganism Toxoid (harmless form of toxin) Preparation of harmless ags

Definition
A vaccination

is an injection of a weakened form of the actual antigen that causes the disease. The injection is too weak to make you sick, but your B lymphocytes will recognize the antigen and react as if it were the "real thing". Thus, you produce MEMORY cells for long term immunity.

Examples of Vaccines
Bacterial vaccines
Live (BCG vaccine for tuberculosis Killed (cholera vaccine) Subunit (typhoid Vi antigen) Bacterial products( tetanus toxoid)

Viral vaccines

Live (oral polio vaccine- SABIN Killed (Injectable polio vaccine- SALK)

Subunit (hepatitis B vaccine)

Combined immunisation

Combination of active & passive methods of immunisation Combined immunisation is to be preferred as in protection of non immune individual with a tetanus prone wound

Vaccination Schedule

Vacccination Schedule

Adoptive immunity
Special type of immunisation is the injection of immunologically competent lymphocytes An extract of immunologically competent lymphocytes, known as transfer factor can be used Treatment of lepromatous leprosy

Local immunity
Concept proposed by Besredka (191924) Importance in the treatment of infections which are localised or where it is operative in combating infection at the site of primary entry of pathogen Special class of immunoglobulins (IgA) forms major component Poliomyelitis, influenza,

Herd immunity
Refers to overall level of immunity in a community Relevant in the control of epidemic diseases When herd immunity is low, epidemics are likely to occur Communicable disease eradication dependant on high level herd immunity

ANTIGEN

Defined as any substance which when introduced parenterally into the body, stimulates the production of an antibody with which it reacts specifically & in an observable manner

Attributes of Antigenicity
Immunogenicity- induction of an immune response Immunological reactivity-specific reaction with antibodies or sensitised cells

Types of Antigens
Complete antigen Hapten Epitope Paratope

Complete Antigen

Able to induce antibody formation Produce a specific & observable reaction with the antibody so produced

Hapten
Greek word Haptein means to fasten Substances incapable of inducing antibody formation by themselves but can react specifically with antibodies\ Immunogenic on combining with large molecular carrier

Epitope
Antigenic determinant Smallest unit of antigenicity That small area on antigen usually consisting of:4 or 5 aminoacid or
monosaccharide residue, specific chemical structure, electrical charge, spatial configuration, capable of sensitising an immunocyte & reacting with its complimentary site on specific antibody or T cell receptor

Epitope

Types of Epitope
Sequential epitope- single linear segment of primary sequence, recognised by T cells Conformational epitope- formed by bringing together on surface residues from different sites during folding into tertiary structure, recognised by B cells

Paratope

Combining area on antibody molecule, corresponding to epitope Epitopes & Paratopes determine the specificity of immunological reactions

Determinants of Antigenicity

Size Chemical nature Susceptibility to tissue enzymes Foreignness Antigenic specificity Species specificity Isospecificity Autospecificity Organ specificity Heterogenetic (heterophile) specificity

Biological classes of Antigens

T cell dependant(TD) T cell independent(TI)
T cell independent
Directly stimulate antibody production by B cells

T cell dependent
Require T cell participation

No participation of T cells

Difference between TI & TD

T cell independant
Structurally simple
Immune response critically dose dependant Too little non immunogenic Much results in immunological tolerance

T cell dependant
Structurally more complex
Immunogenic over wide dose range Do not cause tolerance readily Induce full gamut of immunoglobulin isotype

Antibody response limited to IgM & Immunological memory IgG3 No immunological memory Metabolized very slowly Rapidly metabolized in the body

Remain in body for long periods

Antibodies

Also known as immunoglobulins Globular glycoproteins The heavy and light chains are polypeptides The chains are held together by disulphide bridges Each antibody has 2 identical antigen binding sites variable regions. The order of amino acids in the variable region determines the shape of the binding site

Antibody

Proteins that recognize and bind to a particular antigen with very high specificity. Made in response to exposure to the antigen. One virus or microbe may have several antigenic determinant sites, to which different antibodies may bind.

Structure of Antibody

Structure of Antigen Molecule

Functions of Antibodies
Some act as labels to identify antigens for phagocytes Some work as antitoxins i.e. they block toxins for e.g. those causing diphtheria and tetanus Some attach to bacterial flagella making them less active and easier for phagocytes to engulf Some cause agglutination (clumping together) of bacteria making them less likely to spread

Structure of Immunoglobulins

Immunoglobulin Classes
I. IgG

Structure: Monomer Percentage serum antibodies: 80% Location: Blood, lymph, intestine Half-life in serum: 23 days Complement Fixation: Yes Placental Transfer: Yes Known Functions: Enhances phagocytosis, neutralizes toxins and viruses, protects fetus and newborn.

IgM

Structure: Pentamer Percentage serum antibodies: 5-10% Location: Blood, lymph, B cell surface (monomer) Half-life in serum: 5 days Complement Fixation: Yes Placental Transfer: No Known Functions: First antibodies produced during an infection. Effective against microbes and agglutinating antigens.

IgA

Structure: Dimer Percentage serum antibodies: 10-15% Location: Secretions (tears, saliva, intestine, milk), blood and lymph. Half-life in serum: 6 days Complement Fixation: No Placental Transfer: No Known Functions: Localized protection of mucosal surfaces. Provides immunity to infant digestive tract.

IgD

Structure: Monomer Percentage serum antibodies: 0.2% Location: B-cell surface, blood, and lymph Half-life in serum: 3 days Complement Fixation: No Placental Transfer: No Known Functions: In serum function is unknown. On B cell surface, initiate immune response.

IgE

Structure: Monomer Percentage serum antibodies: 0.002% Location: Bound to mast cells and basophils throughout body. Blood. Half-life in serum: 2 days Complement Fixation: No Placental Transfer: No Known Functions: Allergic reactions. Possibly lysis of worms.

Abnormal Immunoglobulin
Earliest description was the discovery by Bence Jones (1847) Apart from antibodies, they are structurally similar proteins seen in serum in many pathological processes, & sometimes even in healthy persons

Examples
Multiple Myeloma- bence jones protein is a typical finding Waldenstroms Macroglobunemiaexcessive production of M proteins & Bence Jones proteins Heavy chain Disease- lymphoid neoplasia characterized by overproduction of Fc parts of immunoglobulin heavy chain Cryoglobunemia formation of a gel or precipitate on cooling the serum which redissolves on warming

Antigen - Antibody Reactions

Antigens & Antibodies, by definition, combine with each other specifically & in an observable manner In Body- forms basis of antibody mediated immunity In Labs- investigation purposes & surveys In general- detection & quantitation of either antigens or antibodies

General features
Reaction is specific. Specifity is not absolute & cross- reactions may occur due to antigenic similarity or relatedness Entire molecules react & not fragments No denaturation of the antigen or the antibody during the reaction Combination occurs at the surface

General features
Combination is firm but reversible Both antigens & antibodies participate in the formation of agglutinates or precipitates Antigens & antibodies can combine in varying proportions, unlike chemicals with fixed valencies

Affinity
Affinity refers to the intensity of attraction between the antigen & antibody molecules. It is a function of the closeness of fit between an epitope & the antigen combining region of its antibody

Avidity
It is the strength of the bond after the formation of the antigen- antibody complexes It reflects overall combining property of the various antibody molecules in an antiserum

Measurement of Antigen & Antibody

Antigen-Antibody reactions in vitro called Serological Reactions Two important parameters of serological tests are Sensitivity & Specificity Reactions occur in three stagesprimary, secondary & tertiary Reagents & commercially available test kits are used

Sensitivity & Specificity

Sensitivity refers to the ability of the test to detect even very minute quantities of antigen or antibody Specificity refers to ability of test to detect reactions between homologous antigens & antibodies only, & with no other

Precipitation reaction
Abs and Ags in aqueous solutions form a lattice -Precipitin Lattice formation requires: polyvalent Abs Ag must be bivalent, polyvalent

Immunodiffusion
Precipitation reaction in agarose gel 1% agar Reaction is visible Distinct & stable band of precipitation formed Can be stained for preservation

Immunoelectrophoresis
Involves electrophoretic separation of composite antigen into its constituent proteins, followed by immunodiffusion against its antiserum, resulting in separate precipitin lines, indicating reaction between each individual protein with its antibody

Used to detect: a)presence/absence of specific proteins or Ig classes b) immunodeficiency or immunoproliferative disorder

Immunoelectrophoresis

An Ag mixture is 1st separated by charge Then, troughs are cut parallel to direction of electric field and antisera is added to trough Ags and Abs diffuse towards each other to produce precipitin bands

Agglutination reactions

Simple, inexpensive, but sensitive Several types exist:

a) Hemagglutination of RBCs b) Bacterial Agglutination c) Passive Agglutination d) Agglutination Inhibition

Radioimmunoassay

very sensitive test; used for measuring hormones, serum proteins, drugs, etc. at low [C]s ( 0.001ug/ml) measures competitive binding of radiolabelled Ag + unlabelled (test) Ag to high affinity Ab

ELISA
depending on enzyme conugated to Ab reacting with a specific substrate to produce a color rxn. Most sensitive of tests for Ag/Ab!!

Variations of ELISAs: Allows for qualitative or quantitative testing. Each one can be used for qualitative detection of Ag or Ab Also, a std curve based on known [C]s of Ag/Ab can be prepped and an unknown [C} can be ascertained
Indirect ELISA

a.

b.
c.

Sandwich ELISA
Competitive ELISA

ELISA

Immunoflouroscence

Provides a quick method for the id of pathogens and lymphocytes

Abs are conjugated with a fluorescent dye

(fluorescein, rhodamine, phycoerythrin) If Abs bind to specific Ags, they can be illum w/ UV light and emit bright colors There are currently 2 methods employed:
Direct staining Indirect staining

Immunoflouroscence

Complement system

Refers to a system of factors which occur in normal serum & are activated characteristically by antigen- antibody nteraction & subsequently mediate a number of biologically significant consequences

Activities of Complement System

Complement system
Control of inflammatory reaction and chemotaxis Clearance of the immune complexes Cellular activation and antimicrobial defense It is a major effector in immunopathological diseases

Complement activation

Functional protein classes in complement system

Structure & Functions of Immune System

The lymphoreticular system is a complex organization of cells of diverse morphology distributed widely in different organs & tissues of the body responsible for immunity two component concept of immunity Humoral or antibody mediated immunity Cellular or cell mediated immunity

Functions of immune system

Drain fluid from tissue spaces and return to it to the blood Transport materials (nutrients, hormones and oxygen) to body cells Carry away waste products to the blood Transport lipids away from digestive system control of infection

Types of Lymphoid Organs

Primary or central Thymus Bursa of Fabricius Secondary or peripheral lymph nodes spleen mucosa associated lymphoid tissue

Humoral & Cellular immunity

Hypersensitivity

Erythroblastosis foetalis

Penicillin induced Hypersensitive reactions

Delayed hypersensitivity reactions

Summary

Immune response is carried out by white blood cells that circulate between blood and lymphoid organs. Most infectious organisms activate innate immune mechanisms. T and B cells recognize specific antigens and generate cell-mediated and humoral immunity, respectively. The response is regulated by T cells.

Summary

Defects in the immune system result in increased susceptibility to infection, and may themselves cause disease. Understanding these mechanisms is important for control of allergies, autoimmune disease, and organ graft rejection

References

Ananthnarayan & Panikers Textbook Of Microbiology. 7th edition Michael J. Pelczar,JR. Microbiology. 5th edition Goldsby, Kindt, Osborne, Kuby. Immunology. 5th edition Harsh Mohan. Essential Pathology for Dental Students. 2nd edition Encyclopedia for Immunity Internet

Thank you

Thank you