Amanda Smith, Ed.M.Amanda Smith, Ed.M.

Union City High School, Union City, NJUnion City High School, Union City, NJ

AAI Summer Research Program for AAI Summer Research Program for TeachersTeachers

Introduction to the Immune System

Lesson 1

Think back to the Online Stem Cell Lab, health

class, and past science classes.

How many different cell types can you name that are related to the immune system?

List them with their corresponding functions in your notebook.

Do Now

Innate vs. Adaptive Immunity

Innate

Immediate, rapid response

Generalized response, nonspecific

No “memory”

Adaptive

Slow response (a few days)

Specific response to molecules that caused it

Repeated exposure to molecules will cause increased responses

You get the flu vaccine. A few weeks later,

your sibling gets the flu and exposes you to the virus. Which part of your immune system will be prepared to fight off the flu virus?

A. Innate Immune SystemB. Adaptive Immune System

Response Question 1

Found in bone marrow

Give rise to blood and immune cells

Form two lineages: myeloid and lymphoid

Hematopoietic Stem Cells

Myeloid Lineage

Gives rise to eight types of cells found in body tissues, focusing on three1. Mast Cells

Degranulate hormonal mediators when activated, involved with inflammation

Activated by IgE antibodies, physical/chemical injury, or complement proteins

Morphology: typical round cellular shape Defining Organelles: secretory granules and lipid bodies A Visual Interpretation

Image at: http://www.mastcellaware.com/about.html

Myeloid Lineage

2. Macrophages Matured monocytes that migrate into tissues from the

circulatory system Role is to phagocytize dead cells and pathogens

Produce NO, O2-, and H2O2 to break down material.

Specific names for specific locations, ex: Kupffer cells in liver

Morphology: monocyte is a spherical shape, but macrophage shape depends on what tissue it goes and matures.

Defining Organelles: extensive Rough ER, lysosomes/phagosomes, pseudopods

3. Dendritic Cells

Main role is to process antigen material and present it to other cells on its surface.

AKA antigen presenting cell Morphology: form branch like projections (dendrites)

during stage of development Defining Organelles: Golgi, ER,

(proteasome – protein complex that degrades proteins)

Myeloid Lineage

Image at: https://www.boundless.com/microbiology/immunology--2/antigen-presenting-cells/dendritic-cells/

A cut on your arm went untreated, and now, it

is infected by bacteria. Which of the myeloid lineage cells are involved with fighting off the infection?

A. Mast CellsB. MacrophagesC. Dendritic CellsD. A & BE. A, B & C

Response Question 2

Gives rise to three major types of cells1. Natural Killer Cells (NK)

Role: limits the spread of tumors and microbial infections by inducing apoptosis in cells, limiting tissue damage

Morphology: large granular lymphocyte, spherical Defining Organelles: cytoplasmic lytic granules,

secretory lysosomes, golgi, vesicles, microtubules

Lymphoid Lineage

Image at: http://imagebank.hematology.org/AssetDetail.aspx?AssetID=4105&AssetType=Asset

2. B Cells

Role: produce antibodies, become antigen-presenting cells, become memory B cells Memory B cells “remember” specific antigens and

can launch fast immune response if antigen is encountered again.

Live for about 10 years. Morphology: spherical, with very large nucleus

and scant cytoplasm around it, has B-cell receptor on cell surface

Defining Organelles: large ER and Golgi, lysosomes

Lymphoid Lineage

Image at: http://commons.wikimedia.org/wiki/File:Lymphocyte2.jpg

3. T Cells

Several types including T Helper and cytotoxic T cells

Role: secrete cytokines after activation by APC to help aid active immune responses, and destroy virally infected and tumor cells by binding to antigens on their surface, respectively

Morphology: Spherical with T-cell receptor on cell surface, covered with short microvilli

Defining Organelles: ER, Golgi

Lymphoid Lineage

Image at: http://www.iayork.com/MysteryRays/2008/03/30/whats-in-a-name-are-cytotoxic-t-lymphocytes-cytotoxic/

What type of cell is also derived from

hematopoietic stem cells that we have not gone over?

A. Chondroblasts (cartilage cells)B. Erythrocytes (red blood cells)C. Endothelium (cells that line blood vessels)D. Hepatocytes (liver cells)

Response Question 3

Dendritic Cells present antigen to activate

immature T cells to become CD4 or CD8 cells.

TH2 cells and costimulatory molecules

released by TH1 cells activate B cells to

produce antibodies.

Interactions between cells

What are the two major lineages that arise

from hematopoietic stem cells? What types of cells can naïve T cells

differentiate into? What is the relationship between dendritic

cells and naïve T cells? How are T cells related to B cells? What is the difference between an antigen and

an antibody?

Discussion

Graphic Organizer

Show the major lineages and what cells are formed.

Connect lineages with lines and descriptor words.

Start with hematopoietic stem cells.

Review Activity

TEACHER USE ONLY! View diagram showing all cell types derived from hematopoietic stem

cells at http://www.allthingsstemcell.com/category/hematopoietic-stem-cells/

Have students make their own/use worksheet.

Nitric Oxide, Inflammation, and

the Immune Response

Lesson 2

What do you think are some possible effects of

the immune system over-responding (responding too much)?

Would the effects be positive, negative, or neutral? Explain your reasoning.

Do Now

Part of the immune response to pathogens

Caused by vasodilation for the movement of plasma and immune cells into the injured tissues.

Can be acute or chronic. Chronic inflammation occurs if the stimulus for

the inflammation persists.

Mediated by macrophages and released chemicals such as IL-1 and TNF-alpha

Inflammation

Nitric Oxide (NO) is released by macrophages

to destroy the source of inflammation.

NO is produced by the protein inducible nitric oxide synthase (iNOS). iNOS is formed in after injury and inflammatory

mediators are released.

An increase in NO production can lead to tissue injury. Occurs when there’s an increase in inflammatory

cytokines being released

Nitric Oxide & Inflammation

Chronic Asthma

Can be caused by allergies, environmental pollutants, microbial infections, or genetics

NO normally helps to regulate airway vascular tone and pulmonary surface tension. Over expression disrupts innate lung defenses

that modulate lung inflammation, leading to chronic inflammation.

Read over questions for article.

Read “The Inflammatory Response in the

Pathogenesis of Asthma.”

Highlight important information to help you

answer questions as you read.

Complete related worksheet & Inflammatory

Response Flow Chart.

Article Review

Intro to Aseptic Technique & Lab

Procedures

Lesson 3

What does it mean when somebody is septic or

goes into septic shock?

What does Aseptic mean?

How can you practice being aseptic in a lab setting? List as many possible ways as you can.

Do now

Origin: septikos – rotten; a – without

Purpose: to keep the sterile cell culture free of microorganisms from the environment

Four basic ways to achieve this

1. Sterile work area

2. Good personal hygiene

3. Sterile reagents & media

4. Sterile handling

Aseptic Technique

1. Sterile Work Area

Use a cell culture hood.

Maintain a clear work surface.

Disinfect work surface before and after use with

70% ethanol.

Image at: http://www.invitrogen.com/site/us/en/home/References/gibco-cell-culture-basics/cell-culture-equipment/laminar-flow-hood.html

Wash your hands before and after working

with cell cultures.

Tie your hair back.

Wear personal protective equipment.

Gloves

Lab coat

2. Good personal hygiene

Image at: http://web.princeton.edu/sites/ehs/labsafetymanual/sec6c.htm

Commercial materials (bought from a

company) are already sterilized.

Materials prepared in the lab should be

prepared with sterile instruments using

aseptic technique.

3. Sterile Reagents & Media

Wipe outside of containers with 70% ethanol .

Use sterile glass or disposable plastic materials.

Do not cross contaminate.

Do not unwrap materials until the moment you are going to use them.

Cap or seal with tape bottles, well plates, etc. to prevent contamination.

Perform experiments rapidly (but safely!) to minimize exposure to contamination.

4. Sterile Handling

Video

http://media.invitrogen.com.edgesuite.net/Cell-Culture/videos/SterileTechnique.html?CID=ccbvid2

The Micropipette

Used to measure small volumes accurately

Uses disposable tips to prevent cross

contamination

Can be set at a fixed volume or variable – you

specify within a rangeImage at: http://faculty.ksu.edu.sa/moncef/Pages/OperatingtheMicropipette.aspx

Setting the Volume

Turn the volumeter adjustment on top. Stay within the micropipette’s range. Decimal appears only for smaller range

pipettes – make sure to turn the adjustment to the correct volume.

Image at: http://www.medicine.mcgill.ca/physio/vlab/bloodlab/eryfrag2_n.htmTable image at: http://www.scienceteacherprogram.org/biology/JBodenmiller10.html

Drawing liquid1. Push pipette plunger down to FIRST stop while OUT of liquid.2. Place tip into liquid, release plunger to draw liquid up.3. Push back down on plunger to first stop to release liquid.4. Push plunger further to second stop to expel any remaining liquid from tip.5. Release plunger to go back to starting position.

Image at: http://www.karantinapertaniansby.com/en/?hal=detil_artikel&id=2

Learn how to use a micropipette to make a

serial dilution.

Virtual Lab Activity

Practice!

Originally used to count blood cells, now used

for various cell types.

Cells first stained with trypan blue to determine viable from non-viable cells.

Used to calculate amount of cells in a solution from a 10 microliter sample.

Hemocytometer

Image at: http://www.sigmaaldrich.com/catalog/product/sigma/z359629?lang=en&region=US

Hemocytometer virtual lab

Practice!