Vaccines

What is a Vaccine?
A vaccine is a non-pathogenic antigen that mimics a particular pathogen in order to elicit an immune response as if that actual pathogen were in the body. The overall goal of a vaccine is to establish immunity against that particular pathogen.

Mechanism of Vaccination
Establish resistance to virus/pathological organism by evoking an immune response 1. Give host a foreign organism/protein in non-infectious form 2. Antibodies are generated Ab binds to surface proteins of organism 3. Memory B and T lymphocytes Antibody Response Graph

The Mechanism of a Vaccine

In an ideal scenario, whenever a vaccine is first administered, it is phagocytized by an antigen presenting cell (APC). Recent research suggest that it is particularly important that the vaccine be taken up by a dendritic cell. This is because dendritic cells play a key role in activating T cells, which become helper T cells (Th cells).

 From there, the activated Th cells goes on to activate mature B-cells. These activated B-cells divides into two cell types, antibody-producing plasma cells and, most importantly, memory B cells. Memory T-cells are also established, however, they usually have a shorter half-life than memory B cells, thus, they play only a minor role in long-term immunity. Usually, there are no cytotoxic T-cells formed whenever the body responds to a vaccine.

Potential Shortcomings of Vaccines

In some rare cases, a vaccine may directly activate a B cell, without stimulation from Th cells. Such antigens are known as Tindependent (TI) antigens. The problem with such a response is that only Ig-M antibodies are produced and there are no memory cells established. Thus, such a vaccine will be useless against establishing immunity.

 Sometimes, the vaccine may be cleared from the body before it has the chance to properly stimulate the immune system. Some pathogens, particularly viruses, has a tendency to mutate and change there surface antigens, making a vaccine against them ineffective.
This is especially true of malaria, which is constantly changing its surface antigens.

Several different types of pathogens may cause similar infections, thus, several different vaccines may be required for them.
For example, Heamophilus influenzae, a bacterium, and influenzavirus, a virus, causes diseases with similar symptoms.

The Importance of the Secondary Immune Response

During the secondary immune response, the body mounts a quicker, more robust attack on the pathogen. Thus, the pathogen is cleared from the body before it has the chance to cause an infection.

Adjuvants
An adjuvant is a chemical substance that can be added to a vaccine in order to enhance the immune response to the vaccine. There are three types of adjuvants.

Aluminum Hydroxide and Aluminum Phosphate (Alum)

Alum is an inorganic salt that binds to proteins and causes them to precipitate. Whenever the alum/vaccine complex is injected into the body, it slowly dissolves, releasing the vaccine. Bacterial extracts can be added, which enhances the immune response. Alum is the only adjuvant approved for use in humans.

Freund s Adjuvant
In Freund s adjuvant, the vaccine is suspended in oil droplets. When injected into the body, the vaccine slowly diffuses out of the oil drop. Bacterial antigens can be added in order to enhance the immune response. Not used in humans because of risk of severe inflammation.

Routes of Administration
There are three different routs of administration: Intradermal administration.
Three types are intravenous, intramuscular, and subcutaneous.

Oral administration.
Vaccine is usually given in liquid form. Foods, such as tomatoes, have been engineered to produce a vaccine.

Intranasal administration.

 For most vaccines, the immunity against a particular pathogen has a tendency to wear off over time. In this case, a periodic booster administration must be given in order to strengthen and lengthen the duration of immunity. Boosters can also be given during the primary response in order to prolong and strengthen the immune response against the vaccine.

Boosters

Types of vaccines

Traditional
I. Types A. Attentuated (Live, Non-infectious) B. Inactivated (Killed) C. Live LIVE MORE EFFECTIVE THAN KILLED II. Pathogens A. Bacteria B. Virus C. Parasites

Attenuated Virus/Bacteria
These vaccines consist of live, but weakened, viruses or bacteria. These organisms have been altered, either genetically or chemically, in a way that they are not pathogenic. E.g. polio attenuated virus vaccine for yellow fever, which utilizes the YF17D strain, a weakened form of the wild virus.

Advantage:
slow growth allows prolonged exposure to immune system & consequently induce B & T cell response

Disadvantage:
Chance of reversal of virulence

Killed Whole Organism

This vaccine consist of the actual pathogen, however, it has been killed, either by a heat treatment or chemically. Eg. Salk vaccine for polio, inactivated by formaldehyde. rabies vaccine rabies, inactivated by - propiolactane.
Advantage: No danger of mutation or reversal of virulence Disadvantage: Repeated boosters required Inappropriate for CMI induction Chance of reversal of virulence because of improper killing

Live vector Vaccine

The desired gene for antigen is transferred in an attenuated vector Common virusesused Vaccinia, adenovirus Vibrio cholerae
Advantage: multivalent vaccine possible induce both, B & T cell response Disadvantage: Chance of reversal of virulence viral protein causes risk

Toxoids
Some species of bacterial produce what is known as exotoxens. Toxoids are vaccines which consist of exotoxins that have been inactivated, either by heat or chemicals. These vaccines are intended to build an immunity against the toxins, but not necessarily the bacteria that produce the toxins. examples are botulinum antitoxen and diphtheria antitoxen.

Surface Molecules
Proteins, carbohydrates, and lipids, that are found on the surface of pathogens, are isolated and used as a vaccine. Proteins are usually large and complex enough to be used on there own. Carbohydrates and lipids requires to be conjugated with a large protein in order to be immunogenic. An example of surface molecules used as a vaccine are hepatitis B surface antigens.

Anti-Idiotype Vaccines
In this unique type of vaccine, antibodies from a sick individual are isolated. These antibodies are then injected into a lab animal, which may then produce an antibody whose antigen binding site mimics the epitope that the original antibody binds to. These antibodies are then isolated and injected into a healthy individual, who may produce antibodies with an antigen binding site that binds to the antigen binding site of the animals antibodies. Because the animals binding site resembles the epitope of an antigen on a particular pathogen, the individual will have an immunity against that pathogen.

Limitations To Traditional Vaccines

1. can t grow all organisms in culture 2. safety to lab personnel 3. Expense 4. insufficient attentuation 5. reversion to infectious state 6. need refrigeration 7. do not work for all infectious agents

DNA Vaccines
DNA vaccines consist of plasmids that contains genes for certain types of antigens. Once administered, the plasmid is taken up by the target cell and the genes are expressed. The cell then either excretes the antigen or displays it on an MHC-I molecule.

Chimeric Vaccines
Chimeric vaccines usually consist of attenuated viruses that have been engineered to carry antigens from multiple types of pathogens. For example, the yellow fever vaccine YF17D has been engineered to carry antigens from HIV, different types of bacteria, malaria, even cancer. The main of a chimeric vaccine is the establishment of immunity against several different diseases with one administration.

Recombinant Vaccines
1. Subunit Vaccines peptide vaccines Genetic immunization 3. Attentuated Vaccines 4. Vector Vaccines 5. Bacterial Antigen Delivery Systems

Recombinant Vaccines
1. Delete Virulence Genes (can not revert) V/B as Vaccine 2. Clone gene for pathogenic antigen into nonpathogenic virus or bacteria V/B as Vaccine 3. Clone pathogenic antigen gene into expression vector A. Vaccinate with protein 1. Subunit 2. Peptide

Subunit vaccines
Do NOT use entire virus or bacteria (pathogenic agent) Use components of pathogenic organism instead of whole organism Advantage: no extraneous pathogenic particles ie DNA Disadvantage: Is rprotein same as in situ? Cost

Examples of Subunit Vaccines

A. Hepatitis B Problem with Traditional vaccine- HSV is oncogenic envelope glycoprotein D (gD) elicits Ab response Clone gene into vector Express in yeast cells HBsAg - First Recombinant Vaccine (SB)

Examples of Subunit Vaccines

A. HSV Problem with Traditional vaccine- HSV is oncogenic envelope glycoprotein D (gD) elicits Ab response Clone gene for gD into vector Express in mammalian cells Transmembrane protein modify gene to remove TM portion

Examples of Subunit Vaccines

A. Rabies vaccine Problem with Traditional vaccine- Costly envelope glycoprotein (gD) elicits major Ab response Gene optimized for plant expression Suitable signal peptide for glycosylation Cloned gene for RGP into vector Expressed in plant cells Gylcosylated RGP purified and mice immunized Mice protected against live virulent virus infection

Other Subunit Vaccines

B. Tuberculosis Mycobacterium tuberculosis antibiotic resistant strains use purified extracellular (secreted) proteins as Vaccine C. Bordetella pertussis whopping cough express surface antigen in E coli D. Tetanus express toxin in E coli

Peptide Vaccines
Use discrete portion (domain) of a surface protein as Vaccine These domains are epitopes antigenic determinants are recognized by antibodies

Vaccine Production Methods

There are three main vaccine manufacturing strategies:
In-vivo In-vitro Chemical Synthesis

Some vaccines can be produced using any one of the three methods while for other vaccines, only one method will work.

In-Vivo
In in-vivo manufacturing, the vaccine is produced inside a living organism. Embryonated Chicken eggs are are commonly used, particularly in producing flu vaccines. Vaccines, such as anti-idiotype, can also be produced in lab animals, such as mice. There are even some species of plant, such as bananas, that have been genetically engineered to produce a vaccine.

In-Vitro
Here, using recombinant DNA technology, vaccines can be produced in yeast cultures, bacterial cultures, or cell cultures. Recombinant vaccines, such as chimeric vaccines, are produced in this manor. Attenuated virus/bacteria vaccines can also be produced this way.

Chemical Synthesis
Here, instead of using biological systems to produce a vaccine, a vaccine can be produced in a lab. Vaccines that utilize synthetic peptides as well as conjugated lipids and polysaccharides are manufactured this way. Usually, this method is used in combination with either in-vivo or in-vitro production.

Risks Associated With Vaccines

The primary risk associated with vaccines, especially vaccines that utilize live organisms, is that the vaccine itself causes illness. This Happened with the orally administered Sabin vaccine for polio, where some individuals became ill and, in rare cases, even spread the illness to other individuals who were not exposed to the vaccine. Another risk is that the vaccine may behave as a super antigen and over stimulate the immune system. Yet a third risk is that some individuals may have an allergic reaction to the vaccine, especially vaccines produced in embrionated chicken eggs and in transgenic plants.