* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Download ETP: The Immune System
Survey
Document related concepts
Germ theory of disease wikipedia , lookup
Globalization and disease wikipedia , lookup
Lymphopoiesis wikipedia , lookup
Hygiene hypothesis wikipedia , lookup
Childhood immunizations in the United States wikipedia , lookup
Immune system wikipedia , lookup
Molecular mimicry wikipedia , lookup
Monoclonal antibody wikipedia , lookup
Adaptive immune system wikipedia , lookup
Sjögren syndrome wikipedia , lookup
Psychoneuroimmunology wikipedia , lookup
Adoptive cell transfer wikipedia , lookup
Cancer immunotherapy wikipedia , lookup
Polyclonal B cell response wikipedia , lookup
Transcript
ETP: The Immune System Ellen Fasman IISME: Summer 2008 Main Teacher Objective: to enhance student understanding of immunology, as well as interest powerpoint. Students will do a variety of labs, activities, and present a Specific objectives: to explain the structures and functions of the immune system. This includes concepts such as hematopoiesis, innate and adaptive immunity, the inflammatory response, antigen-antibody reaction, and vaccines. Labs will include microscopy using prepared slides of blood smears and pathogens, the ELISA test, blood typing, making serial dilutions, and PCR. Labs will highlight the importance of replicates and controls in an experiment Grade levels: 9th and 10th grade Biology, and grade 10-12 Physiology students . Standards covered: My ETP will focus on Standard #10 of the California State Standards. This includes the following: 10a. students know the role of skin in providing nonspecific defenses against infection. 10b. Students know the role of antibodies in the body's response to infection 10c. Students know how vaccination protects an individual from infectious diseases. 10d. Students know there are important differences between bacteria and viruses with respect to their requirements for growth and replication, the body's primary defenses against bacterial and viral infections, and effective treatments of these infections. 10e. Students know why an individual with a compromised immune system (a person with AIDS) may be unable to fight off and survive infections by microorganisms that are usually benign. 10f. Students know the roles of phagocytes, B-lymphocytes, and T-lymphocytes in the immune system Materials needed: micropipettes with tips, centrifuge, ELISA kits with reagents, antibodies, incubator, freezer, ABO Rh kits Lectures and labs will go together Week 1: Innate and Adaptive Immune Systems Week 2: Structures and functions of the immune sytem, hematopoiesis Week 3: Blood typing and ELISA (antigen/antibody reactions) Week 4: Student projects/assessments 3 PPTs 1. The Immune System 2. The Inflammatory Response 3. Vaccines Topics Covered in PPTs a. Describe the organization of the Immune System and Lymphatic System b. Describe the protective functions of the skin and mucous membranes c. Explain hematopoiesis d. Explain the importance of phagocytes and natural killer cells e. Compare body defenses: non-specific (innate) vs. specific (adaptive) immune systems f. Describe the inflammatory response g. Describe ways in which antibodies act against antigens h. Compare and contrast the development of B and T cells i. State the roles of B and T cells j. Explain how antigen-presenting cells (macrophages) function k. Explain T cell activation and interactions with other cells of the immune response l. Describe how vaccination works m. Explain how the ELISA test works, in addition to its uses, and limitations 4. Assigned Readings: a. Textbook: Biology, Prentice-Hall Publishing Company, 2007. b. Textbook: Essentials of Human Anatomy and Physiology, Marieb, Elaine, Pearson Education, Inc, Eighth Edition, 2006. 5. Activities a. Immune cell posters: B cells, Helper T cells, monocytes, phagocytes, Natural Killer cells, antigen presenting cells, plasma cells(plus antibody group); Groups of 4; jigsaw the parts b. Student PPTs on immune system topics (possible topics: Asthma, Hemolytic Disease of the Newborn, Anaphylaxis, Lupus, Eczema, Crohn’s Disease, allergy, allergy shots, influenza vaccine, smallpox vaccine, poison ivy, HCG pregnancy test, HIV antibody test) 6. Labs: a. ABO/Rh blood typing b. Microscopy: look at stained cells, hematocrit c. Micropipetting and making serial dilutions d. Bodily Fluids Lab (ELISA test) Name: Date: Period: The Immune System Pre-Test 1. What is the body’s first line of defense? 2. What are pathogens? Give three examples. 3. What are the names of the cells of the immune system? 4. In what organs are the cells of the immune system made? 5. What is innate immunity? Give an example. 6. What is adaptive immunity? Give an example. 7. Define antigen. 8. How do antibodies help to defend the body? 9. Differentiate between cellular and humoral immunity. 10. What is a vaccine made of? How does it work? Bonus Questions: Mr. Brown, an 80-year old man, is complaining about having to receive a flu shot every year. Why is it necessary to receive these shots every year? Name: Date: Period: The Immune System Post-Test 11. What is the body’s first line of defense? 12. What are pathogens? Give three examples. 13. What are the names of the cells of the immune system? 14. In what organs are the cells of the immune system made? 15. What is innate immunity? Give an example. 16. What is adaptive immunity? Give an example. 17. Define antigen. 18. How do antibodies help to defend the body? 19. Differentiate between cellular and humoral immunity. 20. What is a vaccine made of? How does it work? Bonus Questions: Mr. Brown, an 80-year old man, is complaining about having to receive a flu shot every year. Why is it necessary to receive these shots every year? Bodily Fluids Lab Objectives To learn: * how to perform an ELISA assay * the use of antibodies in research and in diagnostic/forensic tests * how an infectious agent is spread through sharing of “bodily fluids” * the importance of replicates and controls in an experiment Summary Students use an ELISA assay to determine whether they are carriers of an “infectious agent.” The lab is a simulation; no infectious agents or bodily fluids are used. Background Information The enzyme-linked immunosorbent assay, ELISA, is a test used to detect and quantify specific antigens in a mixture; this mixture could be blood, urine, semen, or other bodily fluids. It is the basis for many diagnostic tests, including the home pregnancy test and the HIV test for AIDS. In the ELISA test, protein (either antigen or antibody) is bound in wells of an assay plate. The assay plate is made of polypropylene and is specifically made for binding to proteins. We will use the antibody-capture assay method. The steps of the general ELISA test are: 1. The sample (bodily fluid) that may contain the antigen of interest is added to a polypropylene plate with wells. 2. The excess sample is removed and wells are washed. 3. Add enzyme-tagged antibodies, which are directed against the antigen of interest. 4. Wash the unbound antibodies. 5. Add a color substrate. 6. View the result: if there is a color change, then the sample contains the infectious agent (antigen of interest). Experimental Procedure Introduction: This experiment contains two parts. In the first part, each student will share his or her “bodily fluids” with three other students. Then each student will perform an ELISA assay on the shared fluids. Each group will run positive and negative controls on a group plate. After recording the data, each student will determine if he or she is infected and the total number of students infected with the “disease agent.” Kit Materials: Solutions: 20X PBS 10% Tween 20 10X Sodium Carbonate Solution Antigen (BSA $ biotinylated-BSA) 1X PBS with BSA Streptavidin-peroxidase 1X Citric Phosphate Buffer TMB 50 mL 10 mL 20 mL 150 mL 10 mL 2 uL 20 mL tablet Abbreviations used in this lab: PBS BSA mL ul mg min TMB H2O2 Ag Ab Tween 20 Phosphate-buffered saline Bovine serum albumin milliliter microliter milligram minute(s) 3,3’,5,5’-tetramethylbenzidine hydrogen peroxide antigen antibody polyoxyethylenesorbitan monolaurate Part 1: Preparing the Samples Materials: Clear “bodily fluids” tubes Yellow microcentrifuge tube (for unshared sample) Blue microcentrifuge tube (for shared sample) Transfer pipette Marker Rack for microcentrifuge tubes Procedure 1. Label a yellow tube and a blue tube with your initials. Divide the “bodily fluids” sample (clear tube) evenly between the yellow and blue tubes using a transfer pipette. 2. Place the yellow tube into the rack at your lab station. This is your unshared sample. 3. Share your sample in the blue tube with another student. To do this, use a transfer pipette to combine the two samples into a single blue tube. Then divide the sample evenly between the two blue tubes. Throw out the transfer pipette. Record the name of the person with whom you share fluids on the form below. 4. Find a second person with whom to share fluids and repeat step #3. Again, record the name and throw the transfer pipette away. 5. Find a third person with whom to share and repeat step #3. Your blue tubes should now contain the “bodily fluids” from 4 different people. You should have 3 names recorded on your form. Record of Sample Sharing Your name: ________________________________ Students with whom you shared “bodily fluids:” 1.______________________________________ 2.______________________________________ 3.______________________________________ Part 2: ELISA Assay on Shared Samples In this part of the lab, you will run an ELISA test on your shared samples. Each sample will be in triplicate and you will share an assay plate with your lab partners. Materials Quantity Mixed “bodily fluids” in blue tube ELISA plate 1 tube per student 1 plate per group Green tube (positive control sample) Pink tube (negative control sample) Orange tube (antibody solution) Purple tube ( color substrate) Squeeze bottle with wash buffer Stack of paper towels 50 uL fixed volume pipette Yellow pipette tips Marking pen Rack for microcentrifuge tubes 1 tube per group 1 tube per group 1 per group 1 per group 1 bottle per group 1 stack per group 1 per group 10 per group 1 per group 1 per student Procedure: 1. Using the 50-uL fixed volume pipette with a new tip, each person in the group adds 50 uL of shared “body fluid” (blue tube) into each of 3 wells. Record the location of each person’s wells on the ELISA assay form. 2. Using a fresh pipette tip for each reagent, prepare 3 positive (green tube) and 3 negative (pink tube) control wells on your plate. Record the location of the controls on the ELISA assay form. Note: the control wells should be adjacent to the experimental wells—not separated by empty wells. 3. Let the plate sit for 15 minutes at room temperature. While the plates are incubating, write your name in the table on the board, as instructed by your teacher. 4. Shake off the fluid into the sink. To empty the wells completely, flick the plates (well side down) against the edge of the sink. 5. Use the squeeze bottle of the wash solution to wash the wells 3 times, shaking out the wash buffer between washes. 6. Tap the plate upside down firmly no a stack of paper towels to remove excess fluid. 7. Using a new pipette tip, add 50 uL of antibody solution (orange tube) to each well. Let the plate sit for 5 minutes at room temperature. 8. Shake off fluid and wash the plate as in steps 4-6 above. 9. Last step: Using a new pipette tip, add 50 uL of color reagent (purple tube) to each well. Let the plate sit for 10 minutes at room temperature. 10. Observe the results. Negative wells (“uninfected” and the negative controls) will remain totally clear. Positive wells (“infected” and the positive controls) will turn blue. 11. Go to the table where you wrote your name and place a plus or a minus next to your name, depending on your results. Also, put a plus or a minus next to the names of the 3 people with whom you shared “bodily fluids.” For example, if your result was positive, you mark a plus sign (+) next to your name and 3 other names and if your result was negative, you mark a negative sign (-) next to your name and 3 other names. Sample Loaded (Student Name) Results (infected/non-infected) ____________________ ____________________ ____________________ ____________________ ____________________ ____________________ ____________________ ____________________ ____________________ ____________________ ____________________ ____________________ ____________________ ____________________ Positive controls ___________________ ____________________ Negative controls ___________________ ____________________ Triplicate Assay Mark (+) or (-) Simple Procedure (at lab bench) 1. Each student receives a clear tube of “bodily fluid.” Put aside half the sample for later testing into a yellow tube. 2. Share your “bodily fluid” with another student by combining them in a single tube, mixing, and splitting them back into 2 tubes again. 3. Now share your “bodily fluid” with 2 more students in the same way. 4. Pour shared sample into a blue tube for testing. 5. To confirm results of the first experiment, the unshared samples can be assayed. Questions 1. What is the full name of the test we simulated in the laboratory? 2. What is the purpose of this experiment? 3. What antibody “tag” does an ELISA use? 4. What’s the difference between a positive and negative test? 5. Why did we run positive and negative controls with the assays? Why did we run them on every plate, instead of just one? 6. What is meant by a “false positive” result? What is one way you could get a false positive result? 7. What is meant by a “a false negative” result?” What is one way you could get a false negative result? 8. Why did we assay each sample in triplicate? 9. Did your 3 wells for each sample have exactly the same intensity of blue color? 10. Why did you wash the plate before you added a new reagent? What could happen if you didn’t wash the plate? 11. How many students in the class were “infected” by the disease agent? 12. How could you figure out who the original “infected” person is? 13. What do your results say about the spread of disease through activities in which bodily fluids are shared? 14. After doing the lab, would you agree or disagree with the following statement: “When you have sex with someone, you are also having sex with everyone that they have previously had sex with.” Explain your answer. Activities 1. Immune cell posters: B cells, T cells, monocytes, phagocytes, Natural Killer cells, antigen presenting cells, Cytotoxic T cells; Groups of 4; jigsaw the parts Poster Requirements: 1. Draw cell and label nucleus, cell membrane, 3 organelles 2. Show the cell in action; it must be interacting with something else (foreign antigen, pathogen, cell, etc). 3. Explain, in steps, the cell in action. What is it reacting to? Specify what it’s doing and if this is an example of innate or adaptive immunity. 4. How is this cell different from all other cells? How is it similar? 2. Student PPTs on immune system topics (Possible topics: Asthma, Hemolytic Disease of the Newborn, Anaphylaxis, Lupus, Eczema, Crohn’s Disease, allergy, allergy shots, influenza vaccine, smallpox vaccine, poison ivy, HCG pregnancy test, HIV antibody test) PPT Requirements: Disease 1. Summary statement: how is this disease significant in the field of immunology? 2. What is the cause of the disease? 3. How does the disease work (symptoms)? 4. Which cells and antibodies of the immune system are involved in the disease? 5. Treatment of the disease Pregnancy, HIV Test: 1. How is this test significant in the field of immunology? 2. What is the reason for the test? When and to whom is it given? 3. How does the test work? 4. Which cells and/or antibodies are involved? 5. What are the limitations and dangers of the test? -Advertisement- Serial Dilutions Made Easy Author: Jan Hilten and Carol Sanders Woodrow Wilson Biology Institute 1993 Introduction: Many areas of science use serial dilutions in the preparations for different experiments. This exercise is presented as an aid for the teacher in helping his/her students improve their skills and more quickly understand the particular application for which serial dilutions are a tool. Serial dilutions are often used in microbiology, biotechnology, and in chemistry classes, to name just a few. Therefore a clear, concise, and non-threatening approach to the learning of this very important concept is essential. Serial dilutions are usually made in increments of 1000, 100 or 10. The concentration of the original solution and the desired concentration will determine how great the dilutions need to be and how many dilutions are required. Important also is the total volume of solution needed. If only small quantities of solutions are needed then greater numbers of dilutions are necessary. The most common examples deal with concentration of cells or organisms, or the concentration of a solute. The approximate concentration should be known at the start of the experiment before the appropriate number and amount of dilutions can be made. In order to arrive at the desired concentration, use serial dilutions, instead of making one big dilution, in order to finally arrive at the desired concentration. This method is not only cost effective but it also allows for small aliquots to be diluted instead of unnecessarily large quantities of materials. This technique involves the removal of a small amount of an original solution to another container which is then brought up to the original volume using the required buffer or water. In the example below, if you have 1 mL of your original solution, and you remove 10 µL and place it in a tube containing 990 µL of water or media you have made a 1:100 dilution. If the original solution contained 5 x 108 organisms or cells/mL, we now have a concentration of 5 x 106 cells/mL, because we have simply divided our concentration by 100. Now, if we want to dilute this by a factor of 1:1000, we must remove 1 µL of the second solution and place it in a tube containing 999 µL of media. We have now diluted our secondary concentration by 1000, and would then divide our concentration by 1000 to yield a 5 x 103 cells/mL. Practice Exercises 1. You are given a test tube containing 10 mL of a solution with 8.4 x 107 cells/mL. You are to produce a solution that contains less than 100 cells/mL. What dilutions must you perform in order to arrive at the desired result? ANSWER: You should perform a series of three 1:100 dilutions to yield 84 cells/mL. 1 mL of original solution to 99 mL of water = 8.4 x 105 cells/mL. 1 mL of second solution to 99 mL of water = 8.4 x 103 cells/mL. 1 mL of third solution to 99 mL of water = 8.4 x 101 or 84 cells/mL. 2. You have a microtube containing 1 mL of a solution with 4.3 x 104 cells/mL and you are to produce a solution that contains 43 cells/mL. What dilutions must you perform? ANSWER: You could perform the following dilutions: 10 µL of original solution to 990 µL of water = 4.3 x 102 cells/mL. 100 µL of second solution to 900 µL of water = 4.3 x 101 or 43 cells/mL. 3. You are given a container with 5 mL of a solution containing 5.1 x 103 cells/mL. You are to produce a solution that contains approximately 100 cells/mL. ANSWER: You would perform the following dilutions: 0.5 mL of original solution to 4.5 mL of water = 5.1 x 102 cells/mL 1 mL of second solution to 4 mL of water = 1.02 x 102 cells/mL or 102 cells/mL 4. You are given a container of yeast cells for which Klett units have been determined on a Klett Summerson Colorimeter. The container contains a population whose concentration is 2.6 x106 cells/mL. You are to prepare a suspension which, when you spread 1 mL of the suspension on appropriate media, will result in about 100 cells. ANSWER: 10 µl of original solution to 990 µl (or 1.0 mL) of sterile water = 2.6 x 104 cells/mL 10 µl of second solution to 990 µl (or 1.0 mL) of sterile water = 2.6 x 102 cells/mL 0.5 mL of third solution to 0.5 mL of sterile water = 1.3 x 102 or 130 cells/mL 0.77 mL of fourth solution to 0.23 mL of sterile water = 100 cells/mL TITLE OF VIDEO: An Inside Look: The Flu VIDEO COMPREHENSION QUESTIONS: 1. What happens immediately after viruses and bacteria enter the body? 2. How does a virus take advantage of the way human cells work? 3. Why does the entire body rather than just the throat experience flu symptoms? 4. How does a T cell become activated, and what happens immediately after it's been called into duty? 5. What are antibodies, and what is their role in fighting the flu? 6. What causes immunity to the flu? An Inside Look: The Flu VIDEO COMPREHENSION QUESTIONS AND ANSWERS: 1. What happens immediately after viruses and bacteria enter the body? When viruses and bacteria first enter the body through the nose, they encounter the hairs lining the nostrils. These hairs trap nearly every particle that's inhaled, and they contain mucus that dissolves bacteria. A virus may survive, however, if it becomes detached from a nose hair and is sucked deeper into the nose. From the back of the nose, the virus can either go to the stomach, where it will be destroyed, or make its way to the throat, beginning the process of giving the person the flu. 2. How does a virus take advantage of the way human cells work? It impersonates one of the proteins that cells use to communicate with each other. A cell can therefore be fooled into thinking the virus is a harmless protein and will permit the virus to enter it. Once inside the cell, the virus begins to use the cell's structure to manufacture components for new viruses. 3. Why does the entire body rather than just the throat experience flu symptoms? As the flu viruses multiply, macrophages in the throat release interleukins, which are chemicals that go through the bloodstream to summon reinforcements to the throat. The interleukins cause nerves to be hypersensitive, making even slight touches uncomfortable. They also raise the body's temperature to make it a less hospitable environment for the virus. This temperature increase tricks the body into feeling cold. Blood vessels around the brain swell, causing a headache. All of these symptoms help remind the flu victim to slow down so that energy can be channeled into defeating the virus. 4. How does a T cell become activated, and what happens immediately after it's been called into duty? Dendritic cells gather fragments of the flu virus and then search the body's lymph glands for an appropriate T cell. Once this T cell has been located, it begins to divide. The new T cells then move to the throat through the bloodstream and selectively destroy the infected throat cells. 5. What are antibodies, and what is their role in fighting the flu? Antibodies are tiny proteins manufactured by B cells. They lock onto the spikes of newly produced viruses, paralyzing the viruses and preventing them from infecting new cells. 6. What causes immunity to the flu? Some T cells remain as memory cells, patrolling the body for the rest of the person's life. If the virus tries to enter the body again, these memory cells will instantly wipe it out, unless it has mutated. Pathogen Project Objectives: to learn about a disease: cause, symptoms, diagnosis, prognosis, mode of transmission, epidemiology, and vaccines (if any) Instructions: Use a variety of resources, including books, periodicals, the Internet, health care professionals, and the local department of public health Below is a list of suggested diseases to research: Polio Pneumonia Botulism Salmonella Meningitis Herpes 1 Herpes 11 Scarlet Fever Tuberculosis Staphylococcus aureus Syphilis Mumps Lyme Disease Encaphalitis Pseudomonas Pyelonephritis Chicken Pox Small Pox Rubella Yellow Fever Rabies Mononucleosis Tetanus Gonorrhea Lupus Anthrax Diphtheria Ebola Streptococcus pyrogenes Format: Poster and speech or powerpoint (including oral presentation) Parts of the Project: 1. Summary Statement: name of disease, standout features, part of the body which is affected 2. Organism causing disease: virus, bacteria, parasite, fungus? Give the genus and species or viral name 3. Symptoms: List the characteristic signs of the disease. Do new symptoms develop as the disease progresses? What happens if the disease goes untreated? 4. Diagnosis: what tests are performed by doctors to diagnose the disease? 5. Mode of transmission: How is this disease passed from one person to another? 6. Treatment: Consider all aspects of treatment, including medication, physical therapy, rest, etc. Mention any treatments which are not based in Western medical tradition; is there evidence for the effectiveness of these treatments? Why do you think they may work? 7. Vaccines: Are they available for the disease? If yes, what type of vaccine is it? What is the recommended schedule for vaccination to prevent this disease? Does the vaccine have any side effects? 8. Epidemiology: Who gets the disease? Mention distribution of disease, based on factors such as age, sex, race, religion, lifestyle, and geographical area. 9. Prognosis: What is the chance of being cured if you contract this disease? If cured, could you get the disease again? Medical Detective A Walk in the Woods Jack is a lawyer who lives in a wooded suburb near Stony Brook, New York He has two children, the youngest being 25 years old. He typically works 50 hours a week. In his spare time he likes to go fishing. His house, a 100 acre estate, is tucked away from the noise and commerce of the city. His daily routine includes going for a walk through the woods and fields of his property with his Golden Retriever, Lancelot. Jack is also an avid birder and usually brings his binoculars with him. . Jack grew up in inner city Pittsburgh in an area where there were a lot of factories and textile mills. Jack had always been healthy but one of his two siblings died of leukemia. The other was chronically ill and the doctors could never come up with a specific diagnosis. In early August, Jack began to feel run down. He noticed that he had been having more headaches than usual. He didn’t go to work for three days due to a fever and general malaise. He felt better for a few days and then missed another two days of work with a low grade fever, night sweats and chills, and severe back pain. Back at work, Jack’s coworkers worried about him. They noticed that he appeared sluggish and without his usual high spirits. They also noted that he was more apt to forget things and had a hard time focusing on important matters at work. Jack quit working because he had worsening pain in his neck and shoulders and he felt too exhausted. He felt a strange weakness in the left side of his body. What was once a small pink dimple on his right leg now looked like a round red rash with a smaller rash in the center. He became depressed and began staying in bed longer and longer periods. Finally, his wife convinced him to see a doctor. Now you be the doctor. What do you think is wrong with Jack? Teacher’s Guide: A Walk in the Woods Disease: Lyme Disease Relevant facts: Lives in wooded area- where there are deer, ticks, mosquitoes Location: NY: site of an epidemic of Lyme Disease Time of year: August: breeding time for ticks Symptoms: malaise, fever, muscle weakness, lack of sensation on one side, depression, outwardly expanding skin rash/bullseye rash (erythema migrans) Medical Detective #2 Kids on Fire Joey is a 5-year old boy who lives with his two brothers in a suburb of Minneapolis. He was born with deafness in his right ear. Joey started kindergarten in the Fall and goes to the school day care center from 12:00 PM until 5:00 PM most school days. It’s a fairly crowded day care, 20 kids on average. Three kids were reported sick this week, all with high fevers. Two days a week, Joey plays soccer at the local elementary school. He had a mild concussion two months prior after heading a ball in a championship game. Joey’s parents both work full-time, and they just had a new swimming pool installed in their backyard, equipped with a hot tub. After a birthday party for his oldest party at their pool, Joey suddenly felt hot. In fact, that night his fever jumped to 103 degrees. After some Tylenol and a lukewarm bath, it went down to 101. However, his mother was still worried. The next morning Joey’s fever spiked again, this time to 105 degrees. He complained of a stiff neck and said there were snakes crawling all over him. Joey’s mother rushed him to the hospital. While he sat in the waiting room, the bright lights made him scream. He said his head felt like it was exploding Sadly enough, a secretary from Joey’s school called to tell her that one of the boys at Joey’s day care suddenly died. Five years later, Joey has remnants of the disease: weakness on one side of his body and loss of the tips of two toes. Now you be the doctor. What do you think is wrong with Joey? Teacher’s Guide: Kids on Fire Disease: Bacterial Meningitis Relevant clues: Goes to daycare with lots of kids, where infections spread easily Two kids reportedly with high fevers, one died Symptoms: high fever, stiff neck, hallucinations (snakes crawling over him), light sensitivity Lasting effects: loss of the tips of his toes/ weakness on one side of his body Medical Detective A Boy Named Chile Chile Juarez is an eighth grader living in Sylvester, Georgia. He is the youngest of three brothers and lives with his mother and grandmother in an apartment above a pawn shop. He likes to play baseball or skateboard at the park with kids who live nearby. He has a stray cat, named Reggie, who came into his life when he was nine years old. Reggie is an outdoor cat who seems to have been in a lot of fights, owning to his numerous scars and “foxhole air.” Chile is an excellent student and his mother dreams of being able to afford college for him someday. She figures by the year 2011, she’ll be able to work fulltime and won’t have her elderly mother to take care of. She works at a local factory which makes truck parts. Chile’s father left when he was only one. When Chile was ten, he came home from school complaining of stomach cramps. After dinner, he experienced more cramps and diarrhea. For the next two days, he lay in bed, and couldn’t hold down any food. His mother did not have medical insurance, so she treated Chile with the best she knew how: cool compresses, Tylenol, and, of course his favorite snack of peanut butter and honey sandwiches. After about one week, his symptoms subsided and he felt his old self again. Four months later, Chile began having terrible pains in his wrists and heels. He stopped urinating. This went on for seven days. Chile’s mother went to Wal-Mart and picked up some aspirin to relieve the pain. His mother brought him to the emergency room after noticing the whites of his eyes turned yellow. At the hospital, his fever spiked to 103.7; Chile’s mother told the doctor about his previous bout with upset stomach and diarrhea. They decided to do a blood and urine test, including a HLA-B27 ELISA, plus a spinal tap. The doctors concluded that he was suffering from complications of his untreated illness six months prior. Now.. you be the doctor…What disease did Chile have? Teacher’s Guide: A Boy Named Chile Disease: Salmonella Poisoning Symptoms: stomach cramps, diarrhea, fever, joint pain, Later symptoms: yellow white of eyes (jaundice), lack of urination (kidney failure) are later effects of untreated Salmonella poisoning—all part of Reiter’s Syndrome Other clues: mother gives him peanut butter sandwiches; there was an outbreak of salmonella contamination in Peter Pan peanut butter from Wal-Mart stores in Sylvester, Georgia in 2007 HLA-B27 ELISA test done- the major test for salmonella poisoning Medical Detective: Bea Stung Bea Hopstone lives in the outskirts of Central Park in New York. She is a freelance writer for the New York Times and the New York Herald Tribune and co-author of History of Dictatorship. She travels frequently to Thailand, where she covers the recent political tension on the Thai-Cambodian border. Born and educated in England, she prefers the rattle and race of New York City to her quiet life in Chelsea. She is 36, a mother of three children, twin girls, aged 4 and a boy, aged 8. Her husband is an architect. Bea enjoys big band jazz, tennis, and long walks with her family through Central Park. She is an excellent ping pong player and regularly plays in tournaments on Canal Street with a team called the Barons of Bounce. She has tendonitis in her right elbow and asthma. Bea was working on an article one sunny September morning in her uptown office, eating a corned beef sandwich with one hand, typing in the other. Over the course of the afternoon, she experienced a headache which wouldn’t stop and nausea. She went home early and lay in bed for what seemed like a couple of hours (It was actually 12). Her family assumed she was exhausted, since she was working more hours than usual, covering a feature story on the recent spike in death rate among crows and blue jays in New York City. Her headache, now much worse, had spread to her back, and her limbs felt especially weak. Her husband took her temperature: 39.4 C. “My head feels like it’s on fire!,” she yelled. He rushed her to the hospital. The doctor, a specialist in tropical diseases, noted the following symptoms: fever, frontal headache, diaphoresis, lymphadenopathy, severe vomiting and diarrhea, bilateral leg weakness, neck stiffness, and severe eye pain. Over the next six days, a broad spectrum antibiotic was administered, which didn’t relieve the most severe symptoms of the disease. A morphine drip through an I.V. was given for pain relief. Below is a summary of the test results for Bea Hopstone: Vital Signs: HEENT: photophobic Blood Pressure: 126/80 Neck: nuchal rigidity Pulse: 96 Respirations: 18 Temperature: 39.4 C, oral Neurological Exam: demonstrates an alert and oriented middle-aged female with normal cranial nerves 1-X11 Test Date Lumbar Puncture 09/04/1999 Results Clear CSF fluid, normal glucose, mildly elevated protein Absence of pathogens using gram stain and culture CBC 09/04/1999 CSF lymphocyte count: 158 cells/uL Total WBC: 10,000 cells/uL Lymphocyte: 4,000/uL Hemoglobin: 11.9g/dL Hematocrit: 33.1% Normal Cranial CT scan 09/04/1999 CSF IgM ELISA 09/10/1999 Toxicology Screen (urine) 09/04/1999 Positive for something else No alcohol or illicit drug use Chest X-ray 09/10/1999 Minimal hyperventilation MRI 09/10/1999 Leptomeningeal enhancement of the conus medullaris and cauda equina Negative for Herpes, AIDS, cytomegalovirus Two days later, Bea became less responsive and her respiratory muscles seemed to be working overtime. She stopped talking and had to be put on a respirator. Now you be the doctor. What do you think is wrong with Bea? Teacher’s Guide: Bea Stung Disease: Viral Encephalitis caused by West Nile Virus Relevant data: increase in mortality rate for blue jays and crows- vectors of West Nile Virus-90% crows killed in 1999 from West Nile Virus New York City- site of West Nile Virus epidemic Bea likes to walk in the park (near mosquitoes, birds) WNV is taken in by mosquitoes, birds; Bea was most likely bitten by an infected mosquito Symptoms: fatigue, fever, joint pain, loss of memory and consciousness, eye pain, high lymphocyte count, clear CSF (It’s milky in bacterial infections Vital Signs: Signs of WNV encephalitis: High pulse High body temperature Nuchal rigidity Photophobia Other clues of WNV viral encephalitis: Antibiotics didn’t work- not bacterial Gram stain and culture-negative--- not bacterial Lab tests: Clear CSF: indication of non-bacterial infection High CSF lymphocyte count (There should be none)- indication of infection CBC: Low RBC, high lymphocyte count MRI shows upper and lower spinal cord swelling ELISA test positive for something else: viral encephalitis Case Study Directions 1. 2. 3. 4. 5. 6. Read the case study a couple of times. Underline what you think are relevant pieces of information. NOT EVERYTHING IS RELEVANT. Make a list of the relevant facts on the Case Study Notes sheet. Research the symptoms. Make a list of “Rule outs” and “Possibilities.” Explain why you rule it out or consider it a possibility. Write a one paragraph essay on which disease you think the patient has. Explain which symptoms support your hypothesis and which symptoms are absent, according to the literature. IT’S OK IF YOUR DISEASE IS INCORRECT. YOU WILL STILL GET CREDIT IF YOUR REASONING IS LOGICAL AND WELL-RESEARCHED. Case Study Notes Relevant Facts/ Patient Background (age, sex, health, lifestyle, place of residence, occupation, etc) Bibliography Symptoms Research on symptoms Rule Outs Possibilities Model of the Immune System Project Objective: to make an interactive model of either Humoral Immunity or Cell-mediated Immunity and LEARN what’s going on Materials use any objects to represent parts of the immune system Parts of the Project: Model: 1. Create a “working” (interactive) model of either Humoral or Cell-mediated immunity. Each member of the group should be able to explain each structure and homeostatic function. Write-up: 2. 3. On a sheet of paper, draw the model. Label each structure and process according to its scientific name. Write a one paragraph summary of how your model works. How is it significant to biology? Directions for Model: 1. Make a moveable model showing the “cascade effect” in either humoral or cellmediated immunity. That is, show how one thing affects another and another down the line. A “domino effect” is one way of thinking about the adaptive immune response. 2. The items in boxes represents different cells. The items on arrows are actions. 3. The structures and actions should be as realistic as possible. Humoral Immunity T Helper Cell - activates B Cell Plasma cell differentiates into Becomes Bacterial cell stimulates differentiates into infects Memory Cell releases antibodies and binds to engulfs and digests Macrophage Bacteria covered with Antigen Cell-Mediated Immunity releases cytokines Antigen-Presenting Cell (APC) Cytotoxic T Cell T Helper Cell activates and activates differentiates infects APC cell Bacterial cell into Infected host cell phagocytosis macrophage T Memory CellMigrates to lymph nodes to be activated quickly during a second invasion References: http://www.accessexcellence.org http://www.biology.arizona.edu Kristi DeCourcy, The ELISA Assay: An Immunological Experiment, Information Manual, Fralin Biotechnology Center, Virginia Tech, 2003 http://www.discoveryschool.com Harlow,E., and Lane,D.1988. Antibodies, a Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor. http://infectious diseases.about.com http://www.informaworld.com, “Biomarkers,” 2007 Elaine M Marieb, Essentials of Human Anatomy and Physiology, Pearson Benjamin Cummings, 2006 Peter Parham, The Immune System, Garland Science Publishing, 2005 www.RnDSystems.com, Human CCL17/TARC, DuoSet ELISA Development System, R&D Systems Europe,Ltd. Acknowledgements: Roche Palo Alto, Mentors: Jennifa Gosling and Mirella Lazarov Roche Palo Alto Staff: Dave Morris PDEI, Dan Cooper PDEI, Rudyard Ress PDEI, Summer IISME Internship, 2008