Vaccine & Vaccination

Veterinary Medicine Brawijaya University

Antibody/ Immunoglobulin
Is family of structurally related glycoproteins produced in membrane-bound or secreted form by B lymphocytes (mature B cells). Membrane-bound antibodies serve as receptors that mediate the antigen-triggered activation of B cells (IgM & IgD) Secreted antibodies functions as mediators of specific humoral immunity by engaging various effectors mechanisms that serve to eliminate the bound

Fab

The antigen-binding regions of antibody molecules are highly variable, and any one individual produces up to 109 different antibodies, each with distinct antigen specificity

ANTIGENS
An antigen is a foreign or recognized as foreign substances, that may be specifically bound by an antibody molecule or T cell receptor Molecules that stimulate immune responses are called immunogens Macromolecular antigens contain multiple epitopes or determinants, each of which may be recognized by an antibody Specificity of an antigen is on its epitopes Linear epitopes of protein antigens may be formed by a sequence of adjacent amino acids, and conformational determinants may be formed by folding of polypeptide chain. Polyvalent antigens contain multiple identical epitopes to which identical antibody molecules can bind.

 Antibodies can bind to two or in case of IgM, up to 10 identical epitopes simultaneously, leading to enhance avidity of antibody-antigen interaction. The relative concentrations of polyvalent antigens and antibodies may favor the formation of immune complexes that may deposit in tissues and cause damage

Requirements for an antigen

Foreign substance for the body Have high molecular weight (> 10.000 Da) Have complex chemical arrangement
Protein Glycoprotein Lipoprotein Polysaccharide

Possess determinant antigen/epitopes

Intrinsic properties & Extrinsic factors influencing the immunogenicity of proteins

Parameter Size Dose Route Composition Increased immunogenicity Large Intermediate Decreased immunogenicity Small ( MW<2.500 Da) Low or High

Subcutaneous > Intraperitoneal > Intravenous or Intragastric Complex Simple

Form

Particulate
Denatured

Soluble
Native Few Differences Rapid release No bacteria Ineffective

Similarity to self protein Multiple differences (level of foreignness) Adjuvants Interaction with host MHC Slow release Bacteria Effective

The dose of antigen used in initial immunization affects the primary & secondary antibody responses
Primary Immunization with different dose of antigen Antibody responses Secondary Immunization with single dose of antigen
Antibody responses

low-zone tolerance

high zone tolerance

Antigen dose

Antigen dose

Booster
First immunization Repeat immunization

Amount of antibody

Days after antigen exposure

Days after antigen exposure

Primary and secondary humoral immune responses

Feature Time lag after immunization Peak response Antibody isotype Primary responses Usually 5 10 days Smaller Usually IgM>IgG Secondary responses Usually 1 3 days Larger Relative increase in IgG and under certain situation s in IgA or IgE Higher average affinity (affinity maturation) Only protein antigens Low doses of antigens, adjuvants may not be necessary

Antibody affinity Induce by Required immunization

Lower average affinity, more variable All immunogens Relatively high doses of antigens, optimally with adjuvants (for protein antigen)

Type of antigen
1. Complete antigen
Immunogenic Induce immunocompetent cells to produce antibodies Reactive The antibodies react specifically to the antigen Ex: Protein, lipoprotein, microbial antigen, etc

2. Incomplete antigen (hapten)

Low molecular weight Immunogenicity : Reactivity : + Hapten-binding protein (protein carrier) Complete Antigen Ex: Antibiotics, drugs, cosmetics

Antigen classification base on:

Genetic
Antigen histocompatibility organs transplantation Auto-Antigen auto immune diseases Iso-Antigen : Individually antigen in a species (blood group) Allo-Antigen Totally different antigen between organism

Dependency to T cell/ thymus

T cell/ thymus dependent antigen (terbalik)
Via B cell to induce antibody LPS, Poly-L-Lysine

T cell/thymus independent antigen

Antigen protein To produce immunity (humoral or and cellular) have to presented by APC to T cell

VACCINE
Edward Jenners (1796) smallpox
Disease Diphtheria Measles Mumps Pertussis Polio

vaccination against

Max number Number Percent of cases (year) cases in 2004 change 206.939 (1921) 895,134 (1941) 152.209 (1968) 265.269 (1934) 21.269 (1952) 0 37 236 18.957 0 -99,99 -99,99 -99,90 -96,84 -100,00

Rubella
Tetanus Hepatitis B

57.686 (1969)
1.560 (1923) 26.611 (1985)

12
26 16 6.632

-99,98
-98,33 -99,92 -75,08

Haemophilus influenzae type B 20.000 (1984)

The success of active immunization in eradicating infectious disease is dependent on numerous factors:
The infectious agent does not establish as latency Does not highly variable of antigenic structure Does not interfere by the host immune response Vaccines are most effective against infections that limited to one species host
Higher antibodies, long-live effectors cells & memory cells

Attenuated & Inactivated Vaccines

Composed of intact non pathogenic microbes:
Its virulence is attenuated Killing Retaining its immunogenicity

The great advantages:

Elicit all innate & adaptive immunity as same as the pathogenic microbes done Inducing protective immunity

Live attenuated usually more effective Viral vaccine often induce long-lasting specific immunity

Purified Antigen (Subunit) Vaccines

Composed of:
Purified antigens from microbes Toxin Usually administrated with an adjuvant

Toxin induce strong antibody responses LPS antigens induce low-affinity antibody responses (T cell independent antigen)
Against Pneumococcal & H influenzae

 Conjugate vaccines induce high-affinity antibody responses Composed of:

Poorly immunogenic antigens Coupling to protein Ex: hapten-carrier conjugates to pneumococcus, H. influenzae, meningococcus

Purified protein induce high-affinity antibody

Stimulate Th & antibody response Not recognized efficiently by class I-restricted CD8+ T cells

Synthetic Vaccines
Identify the most immunogenic microbial antigens or epitopes, to synthesize these in the laboratory synthetic antigens synthetic vaccines Recombinant DNA technology protein in large quantities vaccine made from recombinant DNA-derived antigens
Ex: hepatitis virus, herpes simplex virus, foot-andmouth disease virus

Live Viral Vectors

A vaccine development is to introduce genes encoding microbial antigens in a noncytophatic (not pathogenic) virus and to infect individuals with this virus The great advantage of viral vectors is induce:
Complement CTL responses lyses the target of vaccine kill the infected host cell

Induce both humoral & cell-mediated immunity

DNA Vaccines
An interesting method of vaccination was developed on the basis of an unexpected observation Inoculation of a plasmid containing complementary DNA (cDNA) encoding a protein antigen leads to strong and long-lived humoral and CMI responses to the antigen. It is likely that APC are transfected by plasmid and the cDNA is transcribed translates immunogenic protein that elicits specific responses Vaccine DNA-encoded viral protein eliciting strong CTL responses Vaccine DNA-encoded bacterial protein enhances adaptive immunity

Multivalent vaccines
Problems :
The vaccine poorly immunogenic Induce either humoral or CMI

Approach: 1. SMMA (solid matrix-antibody antigen)

Monoclonal antibody attaching to particulate solid matrices + saturating the monoclonal antibody with desired antigens possible bind mixture peptide/protein contain immunodominant epitopes for T and B cells

2. Desired antigen incorporate into protein micelles/ lipid vesicles (liposomes)/ immunostimulating complexes (ISCOMs), using detergents - Micelles formed by mixing protein antigens in detergent and then removing the detergent - Liposome is lipid bilayer - ISCOM is lipid carrier 3. Membrane protein from various pathogen incorporated into micelles, liposome or ISCOM

Adjuvant
Adjuvants (latin: adjuvare, to help) are substances that when mixed with an antigen and injected with it, enhance the immunogenicity of the antigen. Protect the antigen (vaccine) Non Antigenic material Used to boost the immune response when:
The antigen has low immunogenicity Only small amounts of antigen

 Adjuvant:
Elicit innate immune responses Increase costimulators signals Induce granuloma formation Increase cytokines, such as IL-12 stimulate growth & differentiation of immunocompetent cells Stimulate lymphocyte proliferation non specifically

Mode of action of some commonly used adjuvants

Postulated mode of action adjuvant Prolongs antigen persistence + Enhance costimulatory signal + Induce Stimulates granuloma lymphocytes formation nonspecifically + -

Freunds incomplete adjuvant

Freunds complete adjuvant

Aluminum potassium sulfate (alum) M. tuberculosis B. pertussis LPS Synthetic polynucleotides (poly IC/poly AU)

+
+ -

++
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++
+ + -

+ + +

Passive Immunization
By transfer of specific antibodies Most commonly used for rapid treatment of potentially fatal disease caused by toxin
Tetanus Rabies Diphtheria Snake venom

Naturally transfer maternal antibodies across placenta to the developing fetus Passive immunity is short-lived, because:

Host does not respond to immunization Protection lasts only as long as the injected antibody persists Does not induce memory not protected against subsequent exposure toxin