Microbiology Chapter 22.txt

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tbednarick
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Microbiology Chapter 22.txt
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2010-07-28 16:23:51
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Microbiology Immunity Serology
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Microbiology Chapter 22
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  1. Four ways to acquire immunity
    • Naturally Acquired Active Immunity
    • Artificially Acquired Active Immunity
    • Naturally Acquired Passive Immunity
    • Artificially Acquired Passive Immunity
  2. Naturally Acquired Active Immunity
    Get the disease, then you are immune
  3. Artificially Acquired Active Immunity
    Vaccine
  4. Naturally Acquired Passive Immunity
    • From mother
    • IgA in colustrum
    • IgG in while in the placenta
    • Lasts about 6 months
  5. Artificially Acquired Passive Immunity
    • Given antibodies usually manufactured from horse serum, usually for Hepatitis A
    • Lasts about 6 months
  6. Inactivated viruses
    Cannot cause disease but you usually need boosters
  7. Attenuated viruses
    Weakened virus. Has the possibility of disease but there is a better immune response
  8. DNA vaccines
    • It's getting easier to make these
    • Put together a DNA sequence to create an epitope. The body codes an immune response.
    • Really good
  9. Physical agents that inactivate viruses
    • Heat - DNA & Capsid
    • UV Light - DNA
  10. Chemical agents that inactivate viruses
    • Formaldehyde - DNA
    • Metals - capsid
    • Phenols - capsid
    • Lipid solvents - damage to envelope
  11. Physical and chemical agents can inactivate viruses
    The best is to damage the DNA but not the capsid so you can get a good immune response but not the disease.
  12. Vaccine Strategies
    • Live, attenuated vaccines
    • Inactivated vaccines
    • Toxoid vaccines
    • Subunit Vaccines
    • Conjugate vaccines
    • DNA vaccines
  13. Live, attenuated vaccines contain
    • Weakened microbes that multiply at only low levels, inducing a strong immune response
    • Organisms can revert to a virulent form and cause disease
  14. A single-dose vaccine can
    Combine vaccines for different diseases
  15. Vaccines using attenuated bacteria are
    Difficult and not widely used
  16. Inactivated vaccines contain
    • Killed pathogens, which induce a weaker immune response.
    • Booster shots are required to maintain immunity.
    • They are safer than attenuated vaccines because they cannot cause disease.
  17. Toxoid vaccines contain
    • Inactivated toxins (toxoids).
    • Since the product is inactivated, booster shots are required.
  18. Subunit vaccines contain
    Only those parts of the antigens that stimulate a strong immune response.
  19. Recombinant DNA technology can be
    • Used to create recombinant subunit vaccines.
    • Subunits cannot cause disease.
  20. Conjugate vaccines are created by
    • Attaching bacterial capsule polysaccharides to a toxoid.
    • They elicit a strong immune response
    • HiB vaccine uses the virus capsule. Cooks the carbs and hooks to a toxiod.
  21. DNA vaccines depend on the ability of some cells to
    • Take up and translate foreign DNA.
    • Display the resulting proteins, inducing a strong immune response.
  22. Naked DNA vaccines contain
    • Engineered plasmids that contain a gene from a pathogen.
    • They are not infective or replicative, so cannot cause disease.
  23. Recombinant vector vaccines involve
    • DNA incorporated into an attenuated pathogen.
    • The pathogen takes the DNA into the cells (viral vector) or incorporates the DNA and present antigens (bacterial vector).
  24. Adverse reactions to vaccines are reported to the
    Vaccine Adverse Events Reporting System (VAERS).
  25. People with egg allergies
    Should not take flu vaccinations.
  26. The risk of contracting a disease with death or disability is
    Much greater than any risk associated with vaccines
  27. Adjuvents
    • Make vaccines thick to slow down the bodies ability to destroy the antigen
    • Gives a better immune response
    • Witches brew of mineral oil, aluminum sulfate, cell walls of bacteria
  28. Serological Reactions
    • Precipitation
    • Lattice formation
    • Fluid precipitation
    • Gel precipitation
    • Diffusion and double diffusion
    • Immunoelectrophoresis
  29. Precipitation
    • Requires the formation of a lattice between soluble antigen and antibody
    • Something soluble in liquid
    • Forms a lattice snowflake look
    • The antigen is soluble
    • Need about equal amounts of antigen and antibody to get this effect
  30. Other types of precipitation
    • Fluid precipitation
    • Gel precipitation
    • Diffusion and double diffusion
  31. Determination of titer
    • Looking for antibodies in the blood
    • Use a known amount of antigen and keep diluting
    • Hopefully there is so much antibody, eventually you get a 1 to 1 precipitation
  32. Titer
    Highest dilution that gives a positive result
  33. Gel precipitation
    • Used in determining titer
    • If the line is close to Antigen, there are lots of Antibodies
    • If the line is close to the Antibody, not good immunity to the disease
  34. Ouchterlony plate
    • A gel plate is cut to form a series of holes ("wells") in the gel.
    • A sample is placed in one well, and sera or purified antibodies are placed in another well and the plate left for 48 hours to develop.
    • During this time the antigens in the sample extract and the antibodies each diffuse out of their respective wells.
    • Where the two diffusion fronts meet, if any of the antibodies recognize any of the antigens, they will bind to the antigens and form what is known as an immune complex. This immune complex precipitates in the gel to give a thin white line, which is a visual signature of antigen recognition.
  35. Immunoelectrophoresis
    • Is a general name for biochemical methods for separation and characterization of proteins based on electrophoresis and reaction with antibodies.
    • In immunoelectrophoresis, diffusion is combined with electrophoresis.
    • Pulls antigen based on charge
    • Can separate out lots of antigens.
  36. Gel electrophoresis
    • On a gel-coated slide, an antigen is placed in a central well.
    • An electrical current is run thru the gel to separate antigens by their electrical charge.
    • A trough is made and a known solution of antibody is added.
    • As antigens and antibodies diffuse toward one another through gel, precipitation lines are seen where optimal concentrations of antigens and antibodies meet.
  37. Agglutination
    Clumping when antibodies and antigen are mixed
  38. Passive agglutination
    • Antigens are absorbed onto latex beads.
    • Antibodies are mixed in.
    • Serum antibodies can be detected rapidly by observing agglutination of the carrier particle.
  39. Hemagglitunation
    Red blood cells clumping
  40. Hemagglutination is used to
    • Determine blood type.
    • Detect viruses that cause agglutination of red blood cells.
  41. Blood type can be determined by
    Using antibodies that bind to the A or B blood group in a sample of blood.
  42. Complement fixation
    • Used to detect the presence of either specific antibody or specific antigen in serum
    • Used to diagnose infections, particularly with microbes that are not easily detected by culture methods, and in rheumatic diseases
    • Superseded by ELISA and by DNA-based methods of pathogen detection, particularly PCR.
    • Test serum - Heat to destroy complement
    • Add known antigen & complement
    • If serum is positive
    • then Ag-Ab reaction uses up the complement
    • Else
    • complement is not used
    • Add sheep RBC & anti-sheep Ab
    • If complement exists
    • Then lysys of sheep RBC
    • If lysis then negative
    • If no lysis then posistive
  43. ELISA
    • Enzyme linked immunosorbent assay
    • Requires a positive and a negative control. If either is wrong, throw out and start over
  44. ELISA procedure
    • Hook antigen onto latex bead
    • Add test serum
    • If Ab present then
    • Attach to Ag
    • Add Anti-Antibody
    • If positive, the Anti-Antibody hooks to latex bead
    • The Anti-Antibody has an attached enzyme (horseradish peroxidase)
    • Add substrate � some type of colored protein
    • Enzyme acts on substrate and will change color of substrate if positive
  45. Indirect Fluorescent Antibody Technique
    • For diagnosing syphilis
    • Only looking for antibodies
    • Similar to Eliza but instead of enzymes it uses fluorescence
  46. Production of Monoclonal Antibodies
    • Ab made by plasma cells (working B cells)
    • Fused to myeloma cells from a mouse
    • The fusion results in a clone of hybridoma cells which are immortal and which produce a single type of antibody, the monoclonal antibody
    • Can be used to produce Hep A Ab for those exposed.
  47. Polymerase chain reaction
    • Method to produce multiple copies of DNA
    • Have to have a primer: oligonucleotide
    • Need 15-30 base pairs
    • 1: DNA heated to separate the 2 strands (denature)
    • 2: Cooled to allow primer oligonucleotides to hydrogen bond to each complementary strand
    • 3: The DNA polymerase replicates the new strand by adding the appropriate complementary nucleotide
    • 4: The cycle of heating, cooling and strand replication is repeated many times to produce millions of copies of the same double stranded DNA sequence.
  48. Acquired immunity can result by
    Actively producing antibodies to an antigen.
  49. Active immunity occurs when
    The body's immune system responds to antigens by producing antibodies and lymphocytes.
  50. Naturally acquired active immunity follows
    Illness or pathogen exposure.
  51. Artificially acquired active immunity occurs through
    Vaccination.
  52. Vaccines
    • Contain treated or altered microbes, toxins, or parts of microbes.
    • A primary immune response occurs.
    • Memory cells are formed.
    • The person does not usually become ill.
  53. There are several types of vaccine strategies.
    • Live, attenuated vaccines contain weakened microbes that multiply at only low levels, inducing a strong immune response
    • Organisms can revert to a virulent form and cause disease
  54. Vaccines using attenuated bacteria are
    Difficult and not widely used
  55. A single-dose vaccine can
    Combine vaccines for different diseases
  56. Inactivated vaccines
    • Contain killed pathogens, which induce a weaker immune response.
    • Booster shots are required to maintain immunity.
    • They are safer than attenuated vaccines because they cannot cause disease.
  57. Toxoid vaccines
    • Contain inactivated toxins (toxoids).
    • Since the product is inactivated, booster shots are required.
  58. Subunit vaccines
    • Contain only those parts of the antigens that stimulate a strong immune response.
    • Subunits cannot cause disease.
  59. Recombinant DNA technology
    Can be used to create recombinant subunit vaccines.
  60. Conjugate vaccines
    • Are created by attaching bacterial capsule polysaccharides to a toxoid.
    • They elicit a strong immune response.
  61. DNA vaccines depend on the ability of some cells to:
    • Take up and translate foreign DNA.
    • Display the resulting proteins, inducing a strong immune response.
  62. Naked DNA vaccines
    • Contain engineered plasmids that contain a gene from a pathogen.
    • They are not infective or replicative, so cannot cause disease.
  63. Recombinant vector vaccines involve
    • DNA incorporated into an attenuated pathogen.
    • The pathogen:
    • takes the DNA into the cells (viral vector) or
    • incorporates the DNA and present antigens (bacterial vector).
  64. Acquired immunity can
    Also result by passively receiving antibodies to an antigen.
  65. Naturally acquired passive immunity
    • Ccongenital immunity
    • occurs when antibodies pass from mother to fetus.
  66. Maternal IgG antibodies remain in the child
    3�6 months after birth
  67. Maternal IgA antibodies also pass to the newborn through:
    • First milk (colostrum).
    • Breast milk.
  68. Artificially acquired passive immunity involves
    • Injection of antibody-rich serum into a body.
    • The serum can be used to:
    • Prevent disease (prophylactic).
    • Treat disease (therapeutic serum).
  69. The immune system may recognize foreign serum proteins
    • As "nonself" and mount an allergic reaction.
    • Immune complexes may form and serum sickness may develop.
  70. Herd immunity results from
    • Effective vaccination programs
    • The majority of a population are immune
    • Unvaccinated individuals are unlikely to contact an infected individual.
  71. Herd immunity is affected by:
    • Population density
    • The strength of a person's immune system
  72. The titer is the most dilute
    • Concentration of serum antibody that reacts to its antigen.
    • A rise in the titer ratio (antibody:serum) indicates disease.

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