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HASPI: Measles and Immunity Lab 1

Teacher Information The Vaccination and Immunity Module

Vaccinations are one of our greatest public health achievements. The CDC estimates that 732,000 deaths and 322 million illnesses have been prevented by childhood vaccinations in the U.S. over the last 20 years and $295 billion saved in related direct costs. While vaccines have been highly effective, they may also have become a victim of their own success. Most parents today have never seen the diseases against which we vaccinate, and may not understand their potential severity. Although the documented risks associated with immunizations are low, an increasing number of parents in the US have become fearful of vaccinations and are choosing to delay or refuse vaccines for their children. San Diego County is a good example of this, with an overall rate of personal belief exemption (PBE) from school vaccine requirements for the 2014-‐15 school year of 3.5 percent, higher than the national average and higher than California as a whole. It is important to note that high rates of PBEs from school vaccine requirements cluster within communities, putting the children in these geographic areas at increased risk. For example in San Diego County, PBE rates within individual schools reach as high as 56%.

Dr. Robert Benjamin, Deputy Public Health Office of Marin County stated that, “…a key strategy for improving immunization rates should include educating parents-‐to-‐be, the high school students of today, to understand the science of immunology. Today’s high school science courses should not only teach, but engage students in the real world application of concepts like herd/community immunity, vaccine safety and the importance of immunization throughout the lifespan; thus preparing parents-‐to-‐be to make informed decisions for themselves and their future children”. The vaccination and immunity module is designed to pursue this strategy.

The Vaccination and Immunity module can be used to educate students on the safety, effectiveness, and importance of vaccinations. Students will have the opportunity to watch two documentaries, put themselves in outbreak scenarios, research and discuss vaccination controversies, and educate their peers on vaccine safety and effectiveness. This module has been created and produced by the Health and Science Pipeline Initiative (HASPI) in collaboration with Rady Children’s Hospital, the Jonas Salk Legacy Foundation, the San Diego Rotary, and the San Diego Foundation.

Activity Descriptions

a. A Shot to Save the World Documentary In the early to mid-‐1900s polio was a horrific disease that caused permanent paralysis and that could be fatal. Children were the primary victims. At its peak in 1952, the polio virus caused more than 21,000 paralytic cases in the United States. As a result of widespread vaccination, polio has been eliminated in the United States. “A Shot to Save the World” tells the story of the fear surrounding the polio epidemic, the cooperation among society, politicians, and the scientific community, and the scientists who developed the vaccine that would save thousands of children from paralysis or death. Video questions are included.

b. Measles and Immunity Lab This lab is a simulation of a disease outbreak. Several individuals with measles have been reported at your school. The HASPI Epidemic Prevention Coalition (HEPC) has quarantined your site. No one will be allowed to return home until it has been determined whether a risk of a widespread measles outbreak exists, and whether there are enough individuals on campus who have been vaccinated to prevent an epidemic. If at least 90-‐95% of the campus population demonstrates immunity, an epidemic could be prevented through herd immunity. After analyzing the samples for infection and immunity, the class will determine whether herd immunity exists and whether classmates will be allowed to return home.

c. Vaccination and Immunity Probability Simulation Your class has been exposed to Wookiefacia and you will use probability tools to determine what happens to each patient. Students will calculate the probability of patients being hospitalized or dying and compare vaccinated to unvaccinated probabilities. They will then model what would happen if exposed to this disease with different probabilities of contracting the disease for students with and without immunization.


d. Invisible Threat Documentary An excellent example of how young adults can actively learn the science behind immunizations is the award winning 42-‐minute documentary Invisible Threat made by high school students at Carlsbad High School in California. These students set out to produce a documentary about the human immune system and uncovered a social controversy they didn’t know existed. “Invisible Threat” explores the science of vaccination and chronicles the dangers of not vaccinating through interviews with families, physicians and experts. The film, funded through unrestricted grants from Rotary Clubs in California, has received praise from more than 50 organizations including Children's Hospitals across the country. Dr. Paul Offit, a leading expert and Chief of the Division of Infectious Disease at Philadelphia Children’s Hospital, calls it “one of the best films made on this subject.” Video questions are included.

e. Vaccine Safety and Effectiveness Are vaccinations safe and effective? What are the risks and benefits of vaccinating? In this activity you will have the opportunity to conduct research on vaccines and the vaccination controversy. It is important to note that WHERE you get your research is just as important than WHAT you research. In Part I, you will do directed research to learn more about vaccines and misconceptions, and in Part II you will have the opportunity to do some of your own research into the issue.

f. Vaccination Survey While you have had the opportunity to research and gain more knowledge about vaccination safety and effectiveness, there are many individuals who remain unsure on the issue. In this activity you will conduct a survey to determine the knowledge and opinions of individuals around you in relation to vaccine safety and effectiveness. You will also have the opportunity to share and analyze information from your classmates to get a better idea of how individuals on your campus view vaccines.

g. Discussing the Issue To vaccinate or not to vaccinate, that is the question! You have had the opportunity to learn, research, and perform a survey on vaccine safety, effectiveness, and controversy. Now it is time to discuss the issues with your peers. In this activity, you will discuss the questions, and record/share your answers.

h. Vaccination Education Project Now that you are more educated on the issues surrounding vaccinations, it is time to share what you have learned with your peers. With a partner, you will create an informational poster or brochure to communicate some of the survey results and information you have collected on vaccination safety and effectiveness.

Using the Module

This module can be completed in a science class in a 2 – 3 week period. It is best suited for an Anatomy & Physiology course, but could also be used in a Biology, Medical Biology, or Medical Chemistry course. It could also be utilized as part of a linked learning unit between a Science, English, and Math course. The following schedule is an example of linked learning for this module.

Day 1

Science Math English a. A Shot to Save the World Documentary

c. Vaccination and Immunity Probability

d. Invisible Threat Documentary

Day 2 b. Measles and Immunity Lab

c. Vaccination and Immunity Probability

e. Vaccine Safety & Effectiveness Research – Part I

Day 3 g. Discussion Questions b. Lab Results Analysis and Review

e. Vaccine Safety & Effectiveness Research – Part II

Day 4 -‐ 7 h. Education Project


Lab Materials

Materials will be provided as listed below in HASPI kits. If materials ore no longer available through HASPI, purchasing information has also been provided.

Supply Provided (P) or Needed (N)

Quantity Company/ Item #

Approximate Cost

96-‐Well microtiter plates P 10 Edvotek/666 $19.00/set of 6 Plastic pipettes P 20 Carolina/974512 $7.45 IgM Test (0.04% Bromothymol blue) P 30 ml Carolina/849164 $3.05 IgG Test (Phenolphthalein solution) P 30 ml Carolina/879963 $4.60 0.1 M Sodium hydroxide (or any weak base) P 30 ml Carolina/889551 $5.50 0.1 M Hydrochloric acid (or any weak acid) P 20 ml Carolina/867821 $6.00 Water N 60 ml -‐ -‐ Test tubes N 20 -‐ -‐ Paper towels N As needed -‐ -‐ Paper Sheet (white) N 10 -‐ -‐

Company Contact Information:

Carolina Biological Supply www.carolina.com

800.334.5551

Edvotek www.edvotek.com

800.338.6835

HASPI www.haspi.org

Set Up Information

Measles and Immunity Lab This lab requires quite a bit of set-‐up! All subject fluids and test solutions are simulated, but the IgM and IgG tests are the same tests that would be performed to test for a measles infection and immunity.

Each well on the plate represents 48 patient samples that were taken from a class. Rows A-‐D will be used to test for the IgM antibodies for each patient, and rows E-‐H will be used to test for the IgG antibodies for each patient. For example, the patient samples in well A1 and E1 are from the same patient, but A1 will represent the IgM test and E1 will represent the IgG test.

Prepping the 96-‐well Plates Each well in the plate will need to be pre-‐treated before the lab.

• There are two dropper bottles of solution used to treat the wells: hydrochloric acid (or any weak acid) and sodium hydroxide (or any weak base).

• Rows A-‐D: Adding a drop of hydrochloric acid to any well in rows A-‐D will result in a POSITIVE IgM test, and leaving it empty will result in a NEGATIVE IgM test. The IgM test bottle contains bromothymol blue indicator that turns yellow in the presence of the acid, causing the positive results.

• Rows E-‐H: Adding a drop of sodium hydroxide to any well in rows E-‐H will result in a POSITIVE IgG test, and leaving it empty will result in a NEGATIVE IgG test. The IgG test bottle contains phenolphthalein indicator that turns pink in the presence of the base, causing the positive results.


What Do the Results Mean Table 1 is a summary of the results for both the IgM and IgG tests.

Table 1. IgM & IgG Test Results IgM Test IgG Test Results Positive Negative Early stages of an infection Positive Positive Recent infection or exposure Negative Positive Immunity from vaccination or past infection Negative Negative No immunity and no infection

.

Example of a Completed Plate For the first 5 subjects on this plate the results are:

What to Add to the Wells to Get Results Be careful to only add one drop to each well, and keep the wells from cross-‐contamination. This can skew the results. • High Infection Rate: Infection rate is based on the IgM results. For a high infection rate add a drop of

hydrochloric acid to 4 or more wells in rows A-‐D. If 4 wells are positive then approximately 8% of the class is infected, and if 24 wells are positive then 50% of the class is infected.

• Low Infection Rate: Add a drop of hydrochloric acid to 3 or less wells in rows A-‐D.

• High Immunity: Immunity is based on the IgG results. For a high immunity rate add a drop of sodium hydroxide to 43 or more wells in rows E-‐H (yes, that is most of them). If at least 43 wells are positive, then approximately 90% of the class is immune, or will be immune.

• Low Immunity: Add a drop of sodium hydroxide to 42 or less wells in rows E-‐H.

The specific wells selected do not matter, and they should not be the same for each plate. Each research team will collect the data from one another to compile all of the results. For this reason, you could have a plate with a high infection rate and low immunity, and another with a low infection rate and high immunity.

It is up to each teacher to determine what types of results they would like the students to simulate. In general, if more than 90-‐95% of the samples demonstrate immunity for measles then it is unlikely that an epidemic will occur due to herd immunity. It is up to you whether you would like to simulate adequate or a lack of vaccination. Results can even be different from class to class.

When the lab is complete it is recommended thoroughly rinse the well plates be with soap and water or vinegar to eliminate any solutions or indicator that may remain.

If you are doing this lab in multiple classes, it will save set up time to have the lab groups add the hydrochloric acid and sodium hydroxide to the wells as soon as they are finished with the lab.

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Subject&Wells%

IgM&Test&

Results&+%or%–%(rows&A(D)&

IgG&Test&

Results&+%or%–%(rows&E(H)&

Final&Results&Early%Infection%=%E%

Recent/Exposure%=%R%Immunity%=%I%

No%Immunity%=%NI%

& &&

Subject&Wells%

IgM&Test&


IgG&Test&




No%Immunity%=%NI%

A1/E1& & & & & C1/G1& & & &

A2/E2& & & & & C2/G2& & & &

A3/E3& & & & & C3/G3& & & &

A4/E4& & & & & C4/G4& & & &

A5/E5& & & & & C5/G5& & & &

A6/E6& & & & & C6/G6& & & &

A7/E7& & & & & C7/G7& & & &

A8/E8& & & & & C8/G8& & & &

A9/E9& & & & & C9/G9& & & &

A10/E10& & & & & C10/G10& & & &

A11/E11& & & & & C11/G11& & & &

A12/E12& & & & & C12/G12& & & &

B1/F1& & & & & D1/H1& & & &

B2/F2& & & & & D2/H2& & & &

B3/F3& & & & & D3/H3& & & &

B4/F4& & & & & D4/H4& & & &

B5/F5& & & & & D5/H5& & & &

B6/F6& & & & & D6/H6& & & &

B7/F7& & & & & D7/H7& & & &

B8/F8& & & & & D8/H8& & & &

B9/F9& & & & & D9/H9& & & &

B10/F10& & & & & D10/H10& & & &

B11/F11& & & & & D11/H11& & & &

B12/F12& & & & & D12/H12& & & &!

-‐ + I + + R + + R

-‐ -‐ NI -‐ + I


Name: _______________________________________________ Period: ____________ Date: _____________

Health and Science Pipeline Initiative, Rady Children’s Hospital, The Jonas Salk Legacy Foundation, San Diego Rotary, San Diego Foundation Scenario

This lab is a simulation of a disease outbreak. Several individuals with measles have been reported at your school. The HASPI Epidemic Prevention Coalition (HEPC) has quarantined your site. No one will be allowed to return home until it has been determined whether a risk of a widespread measles outbreak exists, and whether there are enough individuals on campus who have been vaccinated to prevent an epidemic. If at least 90-‐95% of the campus population demonstrates immunity, an epidemic could be prevented through herd immunity.

Since no one is allowed on or off of campus your science class has been recruited to help test students to determine whether they are infected and whether they have immunity from measles. A research team has collected samples from 10 classes. Your class has been separated into 10 research teams that will test the samples for each class. After analyzing the samples for infection and immunity, your class will determine whether herd immunity exists, and if your classmates will be allowed to return home.

What You Should Know About Measles As of 2000, measles was considered eliminated in the United States due to widespread vaccination programs. In the last several years, the safety of the MMR vaccine has been questioned, primarily by a falsified report that the Measles, Mumps, and Rubella (MMR) vaccine caused autism. While we now know this to be false, the public image of the MMR vaccine was damaged. Parents continue to question the safety and effectiveness of the MMR vaccine, and the occurrence of measles outbreaks has increased due to a reduction in the proportion of children receiving the vaccine.

Measles is one of the most highly transmissible diseases and it is difficult to identify infectious individuals who can be contagious for several days before any symptoms develop. In fact, signs and symptoms do not develop until 7-‐18 days following infection. Symptoms of measles include: high fever, dry cough, itchy eyes, light sensitivity, sore throat, runny nose, white spots in mouth, and a rash starting at the face and spreading down the body.


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Testing for Infection and Immunity An antibody test can be used to diagnose a measles infection, confirm whether a person is immune, and detect an outbreak. Our bodies produce specific antibodies for measles in response to an infection or a vaccination. Two types of antibodies are made: IgM and IgG. The IgM antibodies are created shortly after an individual has been exposed or vaccinated. IgM antibodies are present up to for 4 weeks to help the immune system to respond to a measles infection, and then taper off until they disappear. IgG antibodies appear later following exposure/vaccination, but remain in the blood for an extended period of time to prevent the individual from contracting measles. By testing for the presence of both the IgM and IgG antibodies it is possible to tell if an individual is currently infected with measles or mumps, and whether an individual has immunity from a prior infection or vaccination. An individual who is positive for IgM antibodies is recently exposed o currently infected, while an individual who is positive for IgG antibodies is immune. NGSS/Common Core Standards

• HS-‐ETS1-‐1. Analyze a major global challenge to specify qualitative and quantitative criteria and constraints for solutions that account for societal needs and wants.

• HS-‐ETS1-‐3. Evaluate a solution to a complex real-‐world problem based on prioritized criteria and trade-‐offs that account for a range of constraints, including cost, safety, reliability, and aesthetics, as well as possible social, cultural, and environmental impacts.

• HS-‐LS1-‐3. Plan and conduct an investigation to provide evidence that feedback mechanisms maintain homeostasis.

• HS-‐LS2-‐8. Evaluate the evidence for the role of group behavior on individual and species’ chances to survive and reproduce.

• MP.2. Reason abstractly and quantitatively.

• HSN-‐Q.A.1. Use units as a way to understand problems and to guide the solution of multi-‐step problems; choose and interpret units consistently in formulas; choose and interpret the scale and the origin in graphs and data displays.

• HSN-‐Q.A.2. Define appropriate quantities for the purpose of descriptive modeling.

• HSN-‐Q.A.3. Choose a level of accuracy appropriate to limitations on measurement when reporting quantities.


Pre-‐Lab Questions Question Response

1

Why has your school been quarantined?

2

How much of the campus population must demonstrate immunity to prevent a measles epidemic? Why?

3

What has your research team been asked to do?

4

Why have measles outbreaks become more common?

5

Why does the long contagious period of measles make it hard to identify?

6

What are the symptoms of measles?

7

For what can an antibody test be used?

8

What is the difference between IgM and IgG antibodies?

9

What does the presence of IgM antibodies mean?

10

What does the presence of IgG antibodies mean?


Materials

96-‐Well plate Test tubes (2) Plastic pipettes (2) IgM Test White paper Paper towels IgG Test Test tube holder (if needed)

Directions

Testing for the IgM Antibodies Task Response

1

Name of your research team. Research Team Name:

2

Obtain a 96-‐well plate from your instructor. Be careful to keep the well plate upright and DO NOT SPILL any of the contents. Each well on the plate has already been treated with 48 simulated samples. Rows A-‐D will be used to test for the IgM antibodies for each subject, and rows E-‐H will be used to test for the IgG antibodies. For example, the samples in well A1 and E1 are from the same subject.

3 Place the well plate on the white sheet of paper on a flat surface. 4 Label one test tube “IgM” and the other test tube “IgG”.

5

Using a plastic pipette measure 3 ml of IgM test solution and place it in your test tube labeled “IgM.” The plastic pipettes have a one ml mark for measurement (see image).

6

Using the pipette and starting at well A1, add 1 drop of IgM test solution to the well. If the test is negative the well will remain blue. If the test is positive the well will turn yellow. The positive test color can fade, so take note of positive wells as soon as they occur. Color in each positive well on Diagram A to record the results.

Positive Negative

7 Each well represents a different subject. Continue adding 1 drop of IgM test solution to each well in Rows A-‐D and record the results in Diagram A. DO NOT ADD any IgM test solution to Rows E-‐H!

8 Return the remaining IgM test solution in your test tube to the bottle and rinse the plastic pipette.

Testing for the IgG Antibodies Task Response

1 Using a NEW plastic pipette measure 3 ml of IgG test solution and place it in your test tube labeled “IgG.”

2

Using the pipette and starting at well E1, add 1 drop of IgG test solution to the well. If the test is negative the well will remain clear. If the test is positive the well will turn pink. The positive test color can fade, so take note of positive wells as soon as they occur. Color in each positive well on Diagram A to record the results.

Positive Negative

3

Each well in rows E-‐H are from the same subjects as rows A-‐D. For example wells A1 and E1, B2 and F2, and C6 and G6 are for the same subject. Continue adding 1 drop of IgG test solution to each well in Rows E-‐H and record the results in Diagram A.

4 Return the remaining IgG test solution in your test tube to the bottle and rinse the plastic pipette.

1 ml mark

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Diagram A. Record the test results by coloring in the positive wells.

Analyzing the Results Task

What do these results mean? Table 1 below summarizes what the results of each subject’s IgM and IgG test mean in terms of infection and immunity. Use Table 1 to complete the results in Tables 2, 3, and 4.

Table 1. IgM & IgG Test Results IgM Test IgG Test Results

Positive

Negative

Early stages of an infection

Positive

Positive

Recent infection or exposure

Negative

Positive Immunity from vaccination or past infection

Negative

Negative

No immunity and no infection

.

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Summarize the results of Diagram A in Table 2. Use Table 1 to determine the Final Results for each subject. Remember that rows A-‐D and rows E-‐H are for the same subjects. The “Subject” column contains the number/letter for each well.

Table&2.&&&Antibody&Test&Results&&&

Subject&Wells%

IgM&Test&


IgG&Test&




No%Immunity%=%NI%

& &&

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IgM&Test&


IgG&Test&




No%Immunity%=%NI%

A1/E1& & & & & C1/G1& & & &

A2/E2& & & & & C2/G2& & & &

A3/E3& & & & & C3/G3& & & &

A4/E4& & & & & C4/G4& & & &

A5/E5& & & & & C5/G5& & & &

A6/E6& & & & & C6/G6& & & &

A7/E7& & & & & C7/G7& & & &

A8/E8& & & & & C8/G8& & & &

A9/E9& & & & & C9/G9& & & &

A10/E10& & & & & C10/G10& & & &

A11/E11& & & & & C11/G11& & & &

A12/E12& & & & & C12/G12& & & &

B1/F1& & & & & D1/H1& & & &

B2/F2& & & & & D2/H2& & & &

B3/F3& & & & & D3/H3& & & &

B4/F4& & & & & D4/H4& & & &

B5/F5& & & & & D5/H5& & & &

B6/F6& & & & & D6/H6& & & &

B7/F7& & & & & D7/H7& & & &

B8/F8& & & & & D8/H8& & & &

B9/F9& & & & & D9/H9& & & &

B10/F10& & & & & D10/H10& & & &

B11/F11& & & & & D11/H11& & & &

B12/F12& & & & & D12/H12& & & &!


Using the “Final Results” column in Table 2, complete Table 3 with the number and percentage of subjects with an early infection, recent infection or exposure, immunity, and no immunity.

Table 3. Infection & Immunity Data Early Infection

# of Subjects

% of Subjects (divide # of subjects by 48, multiply by 100)

Recent/Exposure Immunity No Immunity

Collaborate with the other research teams to complete Table 4. Other research teams will also be looking to collect this information. Once the numbers from ALL research teams has been collected determine the total number and percentage of individuals in each column.

Table 4. Research Team Data

Research Team Name

# With Early

Infection

# With Recent/ Exposure

# With Immunity

# With No

Immunity

TOTALS

Percentage (divide total by 480,

multiply by 100)


Analysis Questions Question Response

1

Summarize the procedure and results of your research.

2

Why was it important to test for both IgM and IgG antibodies?

3

How much of the tested campus population showed immunity to measles/mumps?

4

According to your results, does enough of the campus have immunity to prevent an epidemic? Explain your answer.

5

Which research teams results showed the highest number of infections (both early & current)?

6

Should classes showing higher amounts of infection remain quarantined from the rest of campus? Why or why not?

7

Explain how the MMR vaccination has prevented (if enough people are immune) OR could have prevented (if there are not enough immune people) a measles epidemic.

8

How would you feel as a student on this campus if this was a real scenario? Why?

Documents

Teacher!Information! TheVaccinationandImmunity!Module!€¦ · HASPI:’’Measles’and’Immunity’Lab’ 2" d.!!Invisible!Threat!Documentary! Anexcellent)example)of)how)young)adults)canactively)learnthe)science