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IMMUNOLOGY LAB

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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

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Immune System

• Lymphoid tissue and Associated Organs:

• Bone marrow• Produces B-cells

• Lymph nodes• Filters lymph

• Spleen• Filters blood

• Thymus• Produces T-cells

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Lymphoid System

• System of mononuclear phagocytes

• Lymphocytes• Macrophages• Dendritic cells • Plasma cells

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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 –

•

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– Second level• Third level

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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

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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

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B-cells Make Antibodies

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Production of Antibodies

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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

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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*

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AB

1stD

3rd

**

I

HF

J

EG

A*

D*

C*

*

*

*

*

*

C*

2nd

source • 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

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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

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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.

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Questions?