Click to edit Master subtitle style 5/9/10 IMMUNOLOGY LAB

5/9/10 Host Defenses 1st defense. Structures, chemicals and processes that work together to prevent pathogens from entering the body – Skin, mucous membranes, microflora, body secretions 2nd defense. Cells, chemicals and processes that work together to destroy pathogens that managed to enter the body – Phagocytosis, antimicrobial chemicals, inflammation and fever 3rd defense. Immune response against pathogens (specific Immunity) – Macrophages and lymphocytes

Click to edit the outline text format Second Outline Level  Third Outline Level Fourth Outline Level  Fifth Outline Level  Sixth Outline Level  Seventh Outline Level  Eighth Outline Level Ninth Outline LevelClick to edit Master text styles 5/9/10 Immune System Lymphoid tissue and Associated Organs: Bone marrow Produces B-cells Lymph nodes Filters lymph Spleen Filters blood Thymus Produces T-cells

Click to edit the outline text format Second Outline Level  Third Outline Level Fourth Outline Level  Fifth Outline Level  Sixth Outline Level  Seventh Outline Level  Eighth Outline Level Ninth Outline LevelClick to edit Master text styles 5/9/10 Lymphoid System System of mononuclear phagocytes Lymphocytes Macrophages Dendritic cells Plasma cells

Click to edit the outline text format Second Outline Level  Third Outline Level Fourth Outline Level  Fifth Outline Level  Sixth Outline Level  Seventh Outline Level  Eighth Outline Level Ninth Outline LevelClick to edit Master text styles 5/9/10 Immunity Adaptive Immunity – Naturally acquired (contact with pathogen) – Artificially acquired (vaccination) Innate Immunity – You are born with Passive Immunity – Transfer of antibodies – Activated T cells from immune host Active Immunity – Antigen –

Click to edit the outline text format Second Outline Level  Third Outline Level Fourth Outline Level  Fifth Outline Level  Sixth Outline Level  Seventh Outline Level  Eighth Outline Level Ninth Outline LevelClick to edit Master text styles – Second level Third level – Fourth level » Fifth level 5/9/10 Innate Vs. Acquired Innate – Non-specific – Responds the same way – Includes barriers (skin and mucosa), chemicals (histamine), and non-specific macrophages Acquired – Memory and specificity – Includes B-cell and T-cell mediated response – Slower response than innate

5/9/10 Immune Response Antigens Surface molecules of pathogens Nonliving toxins, chemicals, drugs, and foreign particles Antibody Primary—produced in response to antigen (ELISA) Secondary— bind to primary antibodies

5/9/10 B-cells Make Antibodies

5/9/10 Production of Antibodies

5/9/10 ELISA – Enzyme linked ImmunoSorbent Assay Qualitative or quantitative Detects antigens or antibodies Amount of colored product from secondary antigen (HRP conjugate) is proportional to antigen-antibody interaction

5/9/10 Simulating a Spread of An Infectious Disease, and Tracking Down this Source Shared fluids will be assayed using ELISA to determine if you have been infected and to track down the source of the infection! Unknown Fluid Sample (A) Sample A + B (mix)→ ½ AB Sample AB + C (mix) → ½ ABC Sample ABC + D (mix) → ABCD *BE CAREFUL OF WHICH SAMPLES YOU CROSS CONTAMINATE*

5/9/10 A B 1st D 3rd * * I H F J E G A* D* C* * * * * * 2nd sourc e In this example, A shares fluids with individual B, then C, then D * = HIV infection You will share fluids with three different people from the neighboring group Infection Simulation

5/9/10 Sandwich ELISA Wells coated with purified antibody Add sample to be tested for antigen Wash of unbound antigen Add enzyme labeled specific antibody Wash off unbound antibody “+” control → known antigen “-” control → omit known antigen

5/9/10 Tracking Down the Infection A B C D E F G + - H After running ELISA, you will write a plus our minus beside your name. You will also mark a plus or minus by the name of the partners you shared Whoever has the most pluses is revealed as the early sources of infections Limitation: source cannot be traced to a single individual.

5/9/10 Questions?