CHAPTER 18   PRACTICAL APPLICATIONS OF

 

                                  IMMUNOLOGY

 

Long before any understanding of the immune system and vaccines had come along, it was recognized that people who had survived a case of an infectious disease were often immune to that disease for the rest of their lives. The first record we have of any type of deliberate exposure to a disease in an attempt to produce immunity was called variolation. This involved injecting pus from a smallpox patient into a healthy person. The result was usually a very mild case of the disease which produced immunity. Unfortunately, some persons treated this way died.

 

Edward Jenner provided the next step, using the cowpox organism to provide immunity to smallpox. This was a much safer procedure.

 

Currently, we are able to produce safe vaccines for many infectious diseases. As a result, smallpox has been eliminated worldwide, and polio and measles are  targeted. Most childhood diseases, which caused numerous deaths as recently as the first half of the 1900’s,  are rare in this country today due to vaccines.

 

Not all diseases can be prevented by vaccines. There is no vaccine for AIDS. No vaccines are available for chlamydias, fungi, protozoa, or helminths. Other vaccines, such as those for cholera and typhoid fever, provide less than complete protection and last a relatively short time.

 

 

TYPES OF VACCINES CURRENTLY USED FOR HUMANS

 

A safe and effective vaccine stimulates the formation of memory B and T cells without causing the actual disease.

 

1. Attenuated whole-agent vaccines—entire microbes which are alive but have been weakened or changed in some way so that they can no longer cause the disease but can still provoke an immune response. These vaccines are generally very effective and often provide lifelong protection. The organisms used often originate from mutations in long-term cultures or may have been treated in some way to weaken them. Examples: Sabin polio vaccine, vaccines against measles, mumps, and rubella (MMR). Rarely, the organisms in this type of vaccine can revert to virulence (back-mutate) and cause the disease.

 

2. Inactivated whole-agent vaccines---microbes have been killed, usually by formalin or phenol. May not be as effective as the vaccines that contain living organisms. Examples: human rabies vaccine, influenza vaccine, Salk polio vaccine, cholera vaccine, pneumonia vaccine.

 

3. Toxoids—these are inactivated toxins, which provoke the production of antibodies that will inactivate the toxin if it is produced by pathogens in the body. These require a series of vaccines and periodic boosters. Examples: tetanus toxoid, diphtheria toxoid.

 

4. Subunit vaccines—these contain only those fragments of the pathogen that best stimulate the immune response, making them 100% safe as far as actually causing the disease. Most are produced by locating the genes for certain proteins found on the outside of the microbe and inserting them into cells by genetic engineering. Cells then make the protein, which is recovered, purified, and serves as the vaccine. These are called recombinant vaccines. Examples:  Hepatitis B vaccine and a new, more effective vaccine for whooping cough. The whooping cough vaccine is not made by genetic engineering, it is still a subunit vaccine, but is made by using parts of disrupted bacterial cells. 

 

5. Conjugated vaccines--used in young children. Immune response to the polysaccharide of bacterial capsules does not work well in very young children. Combining the polysaccharide with a protein allows an immune response even in babies. Haemophilus influenzae type B vaccine is an example.

 

6. DNA vaccines--newest idea in vaccines, not yet in use in humans. Injection of "naked" DNA into muscle tissue of animals results in production of whatever  proteins are coded in that DNA. These proteins stimulate an immune response. In some cases the DNA degrades before achieving the full effect, but these vaccines will most likely eventually become common in the future.

 

 

 

DIAGNOSTIC IMMUNOLOGY

 

 In addition to providing protection against disease,  immunologic functions are widely used today to aid in diagnosis. For this purpose:

          Known antibodies are used to detect the presence of specific antigens

          Known antigens are used to detect the presence of specific antibodies

 

These tests depend on the availability of monoclonal antibodies. These are made by isolating a B cell that makes the desired antibodies and combining it with a cancerous B cell, which will divide indefinitely. The combination is called a hybridoma. These cells can produce unlimited amounts of very pure antibodies, called monoclonal antibodies (Mabs). Monoclonal antibodies are also used in treatment of several diseases.

 

1. Precipitation reactions—soluble antigens react with antibodies to form complexes that settle out of a solution. Immunodiffusion tests are precipitation tests carried out in an agar gel medium. A line of visible precipitate develops in the gel.

 

2. Agglutination reactions—particulate antigens react with antibodies to form visible clumps. Patient’s serum is combined with a known antigen. If the patient has antibodies against the antigen, the reaction will occur.

 

Hemagglutination is the use of red blood cells in an agglutination reaction. This is used for blood typing.

Viral hemagglutination inhibition tests—used for influenza diagnosis.

                             Virus + RBC à Agglutination

                             Virus + RBC + serum with no antibodies à  Agglutination

                             Virus + RBC + serum with antibodies  à No agglutination

 

3. Neutralization reactions—harmful effects of a bacterial toxin or a virus are eliminated by a specific antibody.

 

4. Complement fixation reactions---based on the depletion of a measured amount of complement if an antigen-antibody reaction occurs.

 

5. Fluorescent antibody techniques—a clinical sample is fixed to a slide. Antibodies with a fluorescent dye attached are mixed with the sample. If they find their antigen in the sample, they will combine with it. If not, they will wash off the slide. The slide is viewed—if it fluoresces, the test is positive.

 

6. Enzyme-Linked Immunosorbent Assay (ELISA) tests

    a. Direct—detects the presence of antigens.  Antibodies linked to an enzyme are used.

    b. Indirect—detects the presence of antibodies. Antigens linked to an enzyme are used.