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Applications of Immunology. Talaro Chapter 16. Acquired Immunity. Natural Immunity Acquired as part of normal life experiences Artificial Immunity Acquired through vaccination Active Immunity Results when a person is challenged with Ag that stimulates production of Ab Memory

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acquired immunity
Acquired Immunity
  • Natural Immunity
    • Acquired as part of normal life experiences
  • Artificial Immunity
    • Acquired through vaccination
  • Active Immunity
    • Results when a person is challenged with Ag that stimulates production of Ab
    • Memory
  • Passive Immunity
    • Preformed Ab are donated to an individual
    • Acts immediately but is short term
    • No memory
      • Breast feeding
      • Gamma globulin
      • Monoclonal antibodies
vaccines
Vaccines

Why did the vaccinia virus

work?

  • Provide an antigenic stimulus that does not cause disease
    • Attenuated strain
      • Tissue culture or unnatural / unusual host
      • Hypovirulent
    • Dead whole cells or inactivated viruses
      • Heat, formalin, UV irradiation
    • Purified antigen subunits from cells or viruses
    • Surface antigens produce via rDNA technology
    • DNA vaccines
  • Produces long lasting protective immunity
  • Edward Jenner (page 476)
    • www.sc.edu/library/spcoll/nathist/jenner2.html
    • Cowpox
    • Smallpox
      • Variola
    • Controlled experiments
  • Vaccinia virus
    • Cultured cow pox virus for many years
  • Small pox eradicated in 1973

Immunization using a closely related, less pathogenic organism to give protection against a more pathogenic one.

slide5

Smallpox

www.cdc.gov

Cowpox on human forearm

www.cdc.gov

vaccination success

www.who.int/immunization_safety/en/

www.cdc.gov

  • Small pox
    •  2 million people a year died from small pox until 1967
    • The World Health Organization initiated an immunization
    • campaign that eradicated small pox in 12 years
  • Poliomyelitis (polio)
    • This virus attacks the motor neurons of the brain and spinal cord
    • Causes paralysis and death
    • Immunization campaigns since the 1950s have virtually eradicated
    • polio in developed countries
Vaccination Success

news.bbc.co.uk

slide9

www.cdc.gov

  • Haemophilus influenzae type b
    • Mistakenly believed to have caused influenza
    • Type b strains accounted for majority of bacterial meningitis
  • Meningitis, pneumonia, and epiglottitis
  • Was the leading cause of bacterial meningitis of children 5 years or younger
    • 1/200 childrencontracted Hib
    • Incidence has decreased 99% since the vaccine was introduced

20,000 cases per year in the early 1980’s

1,247 in 2000

slide10

www.who.int/immunization_safety/aefi/immunization_misconceptions/en/index.htmlwww.who.int/immunization_safety/aefi/immunization_misconceptions/en/index.html

Vaccines cause disease

Disease is no longer a threat in my country

Cost of vaccination

Difficulty reaching vaccination center

Not recommended by my physician

Safety concerns

Side effects

SIDS

Autism

Depress the immune system

Mercury Poisoning

Religious beliefs

Herd Immunity

Protection from a disease among unvaccinated individuals occurs when  90% of a population is immunized.

% depends on the disease & vaccine

Content and Design Attributes of Antivaccination Web Sites

Robert M. Wolfe, MD; Lisa K. Sharp, PhD; Martin S. Lipsky, MD JAMA 2002 287:3245-3248.

slide11

Polio Outbreak Occurs Among Amish Families In Minnesota

by David Brown

Washington Post Staff WriterOctober 14, 2005

The first outbreak of polio in the United States in 26 years occurred earlier this fall in an Amish community in central Minnesota, state and federal health officials reported yesterday. Four children have been infected with the virus, although none has become paralyzed. The Amish typically decline to vaccinate their children. The last large outbreak of polio occurred in numerous Amish communities in several states in 1979.

Fears Rising Over Measles Outbreaks

by ROBERT A. HAMILTON

STATE health officials are preparing for what they fear could be ''a major outbreak'' of measles when students who attend Boston University return home next week for spring break. Last week, a measles case was reported in Fairfield, an 18-year-old Boston University freshman who returned home eight days ago with cold symptoms and by Tuesday was in the hospital. ''We expect a lot more students will be returning to Connecticut next week, for the spring break,'' the program director of the state immunization program, Dr. Charles H. Alexander, .

March 3, 1985 The New York Times

slide12

irradiation

hypovirulent

serology

Abs

Proteins

Clotting factors

Hormones

Nutrients

Ions

Serology

Clear fluid

from clotted

blood

leukocytes

  • A part of immunology that attempts to detect signs of infection in a patient’s serum
    • Use Abs that specifically bind to Ag
  • Ag-Ab reactions are visible by
    • Clumps
    • Precipitates
    • Color changes
    • Release of radioactivity
  • The most effective tests have high specificity and sensitivity.

Isolate Ab from serum

agglutination test

Isolate Abs from

patient

Agglutination Test
  • Ab and Ag from a whole cell crosslink
    • Forming complexes that settle out and from visible clumps in the test chamber
precipitation tests
Precipitation Tests
  • Soluble Ag is precipitated
    • Cloudy or opaque zone
    • Many variations are used to maximize this technique
    • Measure optical density
slide20

Calculate titer

The highest dilution of serum that shows a positive result

enzyme linked immunosorbent assay elisa
Detect unknown Ag or Ab

A positive result is visualized when a colored product is released by an enzyme-substrate reaction

Enzyme-linked immunosorbent assay (ELISA)

Anti-immunoglobulin Ab complexed to an enzyme

conjugate

slide22

Perform a virtual ELISA courtesy of the Howard Hughes Medical Institute

www.hhmi.org/biointeractive/vlabs/

slide23

Radioimmunoassay

Ag or Abs are labeled with radioactive isotopes and traced

elisa www biology arizona edu
ELISAwww.biology.arizona.edu

Positive Rxn

Negative Rxn

+ is 0.5 or greater

Indeterminate

0.300 – 0.495

- is 0.300 or less

western blot

Western Blot for a HIV on page 487

Western Blot
  • More sensitive than ELSIA
    • Less chance of a false +
  • Detects specific Ag or Ab proteins in serum
  • Separates proteins into bands via gel electrophoresis
  • Protein bands are transferred to a membrane
  • Specific Abs are used to “probe” the membrane
    • Primary Ab are from patient
    • Secondary Ab
      • Anti-immunoglobulin Ab complex to an enzyme
    • Colorless substrate
  • Colormetric
western blot for hiv www biology arizona edu
Western Blot for HIVwww.biology.arizona.edu

Serum proteins (virus) separated via electrophoresis.

Transferred to a membrane.

The primary Abs from the serum of a patient is added and will react to the HIV proteins (above).

The secondary Ab is an anti-immunoglobulin conjugated to an enzyme. This Ab is specific for the primary Abs.

A colorless substrate is cleaved if the anti-immunoglobulin conjugated to the primary Ab.

slide27

Band Pattern Interpretation

Lane 1, HIV+ serum (positive control)

Lane 2, HIV- serum (negative control)

Lane A, Patient A

Lane B, Patient B

Lane C, Patient C

Patient A is –

Patient B is –

Patient C is +

slide29

Immunofluorescence

  • Fluorescent Ab (FAbs) either directly or indirectly to visualize cells or cell aggregates
monoclonal antibodies mab
Monoclonal Antibodies (Mab)
  • Single specificity antibodies formed by fusing a mouse B cell with a myeloma cell
    • A malignant tumor formed by the cells of the bone marrow
  • Used in diagnosis of disease, identification of microbes and therapy
slide31

Hybridomas produce antibody that

  • recognize single epitope
  • Produce uniform, highly specific
  • Ab in large supply
  • Multiple practical applications
  • Diagnostic tests
    • ELISA & Western Blots
  • Immunosuppressive therapy for
  • transplants
    • Prevent action of TC
  • Anticancer drugs
  • Antitoxins
  • Identify a pathogen
  • Purification of an important protein
    • Precipitate
  • Immunize animal
  • Harvest spleen
  • Fuse B & myeloma
  • Hybridoma
    • Immortal
  • Screen hybridomas for Abs
  • directed against antigen of
  • interest
monoclonals on the market
Monoclonals on the Market

Herceptin

Blocks the effects of the growth factor protein HER2, which transmits growth signals to breast cancer cells. Causes tumor shrinkage.

Mylotarg

Acute myeloid leukemia (AML)

Rituxan

Treat a variety of non-Hodgkin's lymphomas.

Target and destroy non-Hodgkin's lymphoma cells.

Remicade

Auto-immune disorders like Crohn's disease and rheumatoid arthritis

Target is tumor necrosis factor

Zenapax

Prevent rejection in organ transplantation, especially in kidney transplants.

Additional information at www.fda.gov