Unit 5: Vaccines and Immunodiagnostics

Table of Contents

1. Vaccines and Vaccination

A vaccine is a biological preparation that provides active acquired immunity to a particular disease. It "teaches" the immune system to recognize and fight a pathogen without causing the disease itself.

Vaccination is the act of administering the vaccine. The goal is to stimulate the adaptive immune system to create immunologic memory (memory B-cells and T-cells). When the real pathogen attacks, the secondary response is so fast and strong that the person does not get sick.

2. Passive and Active Immunization

a) Active Immunization

b) Passive Immunization

3. Types of Vaccines

Vaccine Type Description Advantages Disadvantages
Live-Attenuated (Bacterial/Viral) A "weakened" version of the living pathogen that can still replicate but not cause disease. Very strong, long-lasting immunity (activates both cellular and humoral). Can revert to virulent form (rare). Cannot be given to immunocompromised.
Inactivated (Killed) (Bacterial/Viral) The pathogen has been killed (e.g., with heat or chemicals) and cannot replicate. Very safe, cannot cause disease. Weaker response (mainly humoral). Requires multiple doses ("boosters").
Subunit / Recombinant Contains only a *piece* (antigen) of the pathogen (e.g., a protein). Made using recombinant DNA technology. Extremely safe, no risk of infection. Requires adjuvants and boosters.
Toxoid An inactivated version of a bacterial *toxin*. Teaches the body to fight the toxin, not the bacteria. Effective against toxin-mediated diseases (e.g., Tetanus, Diphtheria). Requires boosters.
DNA / mRNA Vaccines Injects the genetic *instructions* (DNA or mRNA) for the antigen. The host's own cells make the antigen. Fast to develop. Elicits strong cellular and humoral response. Newer technology; mRNA requires ultra-cold storage.

4. Adjuvants and Cytokines

a) Adjuvants

An adjuvant is a substance added to a vaccine (especially non-live vaccines like subunit) to enhance the immune response. It acts as a "danger signal," stimulating the innate immune system and attracting immune cells to the injection site, which makes the adaptive response stronger and longer-lasting.

A common example is Alum (aluminum salts).

b) Cytokines

Cytokines are small, soluble proteins that act as signaling molecules for the immune system. They are the "messengers" that cells use to communicate. (e.g., "Activate!", "Divide!", "Come here!", "Stop!").

Examples include Interleukins (ILs) (which communicate between leukocytes) and Interferons (IFNs) (which interfere with viral replication).

5. Introduction to Immunodiagnostics

Immunodiagnostics are laboratory techniques that use the high specificity of the antibody-antigen reaction to detect and/or quantify a specific substance (e.g., a hormone, a drug, a viral protein) in a sample (e.g., blood, urine).

The core principle is: If you have a known antibody, you can detect an unknown antigen. If you have a known antigen, you can detect an unknown antibody.

6. ELISA (Enzyme-Linked Immunosorbent Assay)

ELISA is a very common technique used in pregnancy tests, HIV tests, and many research labs.

Principle: It uses an antibody that is covalently linked (stuck) to an enzyme. When the correct substrate is added, this enzyme produces a measurable color change.

[Image of sandwich ELISA procedure]

Example: Sandwich ELISA (to detect an antigen)

  1. Coat: A plastic well is coated with a "capture" antibody (specific for the antigen we're looking for).
  2. Sample: The patient's sample (e.g., blood) is added. If the antigen is present, it gets "sandwiched" and stuck to the capture antibody.
  3. Detect: A second "detection" antibody (also specific for the antigen) is added. This antibody is linked to an enzyme.
  4. Substrate: A colorless substrate is added. If the enzyme is present (meaning the antigen was "sandwiched"), the substrate is converted to a colored product.
  5. Result: Color change = Positive test. No color change = Negative test.

7. RIA (Radioimmunoassay)

RIA is an older, highly sensitive technique used to measure the concentration of very small molecules (like hormones).

Principle: It is based on competition.

  1. Setup: You start with a known amount of "hot" (radioactively labeled) antigen and a limited number of "cold" (non-labeled) antibodies. They all bind together.
  2. Sample: You add the patient's sample, which contains an *unknown* amount of "cold" (non-radioactive) antigen.
  3. Competition: The "cold" antigen from the patient *competes* with the "hot" antigen to bind to the limited antibodies.
  4. Result: The *more* "cold" antigen there is in the sample, the *less* "hot" antigen will be able to bind. By measuring the final radioactivity, you can calculate the concentration of the unknown antigen.

8. Immune-electrophoresis

This technique combines two methods to separate and identify proteins in a complex mixture (like blood serum).

  1. Step 1: Electrophoresis: The serum sample is placed in an agar gel and an electric current is applied. The proteins separate based on their charge and size (e.g., albumin, alpha, beta, and gamma globulins).
  2. Step 2: Immunodiffusion: A trough is cut in the gel parallel to the separated proteins. An antiserum (containing antibodies against all serum proteins) is added to the trough.
  3. Step 3: Precipitation: The separated proteins and the antibodies diffuse towards each other. Where they meet, they form visible precipitin arcs (lines of precipitate).

Use: It can be used to identify if a person is missing a specific protein or is over-producing an abnormal one (e.g., in multiple myeloma).