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Applications of Immune Response
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Terms in this set (52)
Natural immunity
Antibodies are acquired from the mother though breastfeeding.
transport gut biome.
IgG: from mother's placenta.
IgA: from mothers breast milk.
EXAMPLE: chicken pox
Artificial immunity
by injection of antibodies.
makes the individual at high risk/immunocompromised and they cannot make gamma globulins IgG.
EX: rabies, tetanus, Hep. A, Hep. B, measles, chicken pox, E. Bola.
Effective 2-3 months of age.
Transfer antibodies produced by another person or animal. Can be used to prevent disease before or after likely exposure.
passive immunity
NO MEMORY.
-naturally acquired: antibodies acquired from the mother through breastfeeding. (ex: chicken pox)
-Artificial or induced: by injection of antibodies. (ex: ebola)
Naturally:
-IgG from mother crosses placenta, infers protection to the baby.
-IgA antibodies in the breast milk given to the child.
active immunity
set-up memory to antibodies through stimulation by infection or vaccination.
Natural: infection, contact with pathogen. (ex: fighting off the cold)
Artificial: injection of immune serum, gamma globulin. (ex: injection of flu virus to build up resistance to a disease)
Attenuated vaccines
weakened form of a pathogen generally unable to cause disease.
Strain replicates in vaccine recipient: causes infection with undetectable or mild symptoms.
RESULT: long-lasting immunity.
virus genes for virulence are removed/weakened. Replicates but slowly; upon maturation may revert to virulence in small cases, pregnancy and immunocompromised.
EX: FLUAD, H1N1 (swine flu), Sabin poilo vaccine, MMR, rotavirus, shingles, yellow fever, chicken pox zoster.
Advantages of Attentuated vaccines
only required single dose (usually) to induce long-lasting immunity.
Vaccine has added potential for being spread to unimmunized individuals.
Disadvantages of attenuated vaccines.
have potential to cause disease in immunocompromised individuals.
pregnant women should avoid immunization as well.
Inactivated vaccines
unable to replicate in vaccinated individual. retains immunogenicity of infectious agent - NOT PATHOGENIC but IMMUNOGENIC.
Falls into two categories:
1. whole agents.
2. fragments.
Whole agents
contain killed organisms of inactivated virus.
does not change epitopes.
EX: cholera, plague, influenza, stalk polio.
Fragments
portions of organisms or agents including toxins, proteins, and cell wall components (PAMPS).
Includes toxoids, protein subunit vaccines, and polysaccharide vaccines.
Toxoid vaccines
inactivated toxins but retain antigen determinants on the molecule.
EX: diptheria and tetanus.
empty viral capsids: polysaccharides/capsules, conjugate.
Pertussis
Vaccine type: subunit.
vaccine in two forms: Whole-cell vaccine (wP) or the acellular vaccine (aP).
part of DPT vaccine unit.
vaccines given to baby in first few months.
TOO MUCH.
the overall effects are not known on the immune system.
Hepatitis B
Vaccine type: recombinant
Rotovirus (DPT)
Vaccine type: attentuated
Diptheria
Vaccine type: toxoid.
Tetanus
vaccine type: toxoid.
Hib
vaccine type: conjugate (2 Ag bound: weak polysaccharide and stonger protein).
Influenza
non-attenuated vaccine--> Inactivated whole agent vaccine.
Previously would use the nasal swab/mist, attenuated: ineffective, low efficacy especially in children.
2017-2018
QUADRIVALENT VACCINE: protects against 4 strains of influenza vaccine.
what diseases do we have vaccinations for?
Cholera
Dengue
Diphtheria
Hepatitis A
Hepatitis B
Hepatitis E
Haemophilus influenzae type b (Hib)
Human papillomavirus (HPV)
Influenza
Japanese encephalitis
Malaria
Measles
Meningococcal meningitis
Mumps
Pertussis
Pneumococcal disease
Poliomyelitis
Rabies
Rotavirus
Rubella
Tetanus
Tick-borne encephalitis
Tuberculosis
Typhoid
Varicella
Yellow Fever
What diseases do not have vaccines?
HIV/AIDS
Malaria
Hookworm infection
herpes simplex virus
Group B streptococcus
shigella
staphylococcus aureus
streptococcus pneumoniae
tuberculosis
What diseases have been eradicated?
only one in humans: small pox
What diseases are under 'control'?
diptheria (86%)
polio (86%)
whooping cough, aka pertussis (64%)
measles (60%)
neonatal tetanus (58%)
...
What diseases are still a major problem?
Hepatitus B, tuberculosis, malaria, HIV, Diarrhoea/enteric fevers, Acute respiratory infections
...
Immunoassay
Identify unknown antigens and/or detect specific antibodies.
obtain an antibody by harvesting the animal's serum.
serological test.
titer.
adjuvant
pharmaceutical/immunological agent that modifies the effect of other agents.
can be added to a vaccine to give higher amounts of antibiotics and longer protection; minimizes the amount of injected foreign material.
serology
method used to determine the presence of antibodies.
tests to study antibodies and
antibody/antigen interactions in blood serum.
herd immunity
when a significant portion of a community is immunized and protects those who are not immunized from infection.
chains of the infection are stopped.
important method of protection for those individuals who cannot be immunized. can become dangerous is un-immunized individuals leave their immunized herd to be around other unimmunized individuals-- results in the spread of a disease.
anti-human IgG antibiotics
antibodies that bind to human IgG; termed anti-human IgG.
can be produced in animals immunized
with IgG from human serum (foreign antigen to
that animal)
Seronegative:
• Person not yet exposed to antigen and has
no specific antibodies.
Seropositive:
• Person with exposure and actively
producing antibody.
Titer:
• Concentration of antibody in serum---
Indicates previous exposure.
How are antibodies obtained?
Plasma is fluid portion with clotting factors; Serum is fluid portion of blood with no clotting factors.
Laboratory animals are used to produce known antibodies: Animal is immunized with antigen and produces specific antibodies.
Antibodies are retrieved by harvesting animal's serum.
Quantifying antigen-antibody
rxns
Concentrations of antibody usually
determined through dilution.
Antigen added to dilution; Titer is taken from last
dilution to give detectable
reaction.
Antigen-antibody complexes form _________ or binding complexes.
aggregates
Antigen-antibody binding can be seen in_______(settling out of
solution) and _________ (clumping) reactions.
precipitation, agglutination
Precipitation reactions.
soluble proteins become insoluble.
Antibodies binding to soluble antigen form insoluble complexes that precipitate out of solution.
Complete aggregate formation occurs at certain
concentrations; To achieve concentrations, place separate antigen and antibody suspensions side by side.
Involve soluble
antigens with
antibodies diffusing
and thus precipitating
when equal titers of
each occur.
Agglutination reactions
particles settle out of suspension.
Reactions involve particulate (insoluble antigens larger than those used in PPT rxns) and antibodies. Antigens may be on a cell such as a Red Blood Cell (direct agglutination); Ab attached to latex beads (indirect or passive agglutination results from binding of Ag).
Zone of equivalence
a.k.a. zone of optimal proportion.
in this zone the antibody and antigen molecules mix and are extensively cross-linked.
precipitation occurs.
Immunodiffusion tests
PPT reactions carried out in an agar gel medium.
Most widely known is Ouchterlony technique.
Antigen and antibody placed in separate wells cut in gel; Solutions diffuse and meet between the wells.
Result in line of precipitation at zone of
optimal proportion
Radial immunodiffusion test is quantitative
1. Antibody is added to liquid agar that is allowed to harden; Creates a uniform antibody concentration in agar gel.
2. Antigen added to wells cut in the agar gel
Diffusion outward of Ag forms concentration gradient; Ring forms at antigen-antibody precipitation.
Standards can be used to construct standard curve to establish concentration.
Immunoelectrophoresis
A combination of imunodiffusion and electrophoresis techniques.
1. Proteins separated using gel electrophoresis.
2. Antibodies are placed in wells and allowed to diffuse towards separated proteins.
3. Line of precipitation forms at antibody-protein recognition.
Used to determine patient antibody level.
DIRECT agglutination tests
• Specific antibody mixed with insoluble antigen.
• Readily visible clumping indication of positive result.
Used in Blood Typing with RBC's and Ab.
INDIRECT agglutination
Amplifies aggregation formation.
1. Ag are adsorbed onto particles of clay or latex beads.
2. Antibody attaches to antigen adhering to the latex beads.
3. Agglutination of these beads occurs.
May test for Ab or Ag.
Fluorescent antibody test
Relies on fluorescent microscopy to locate fluorescent-labeled antibodies bound to antigens that are fixed to a microscope slide.
Direct FA test
Indirect FA test
DIRECT Fluorescent antibody test
Used to identify an unknown Ag fixed to the slide.
Known Ab/unknown Ag binding will be visible under fluorescent microscopy and will ID the Ag.
Unbound Ab is washed away;
Detect unknown Ag complexed with Ab.
INDIRECT Fluorescent antibody test
Used to identify the presence of a given Ab in serum. Known Ag fixed to the slide. Add serum with unknown Abs. Add Fluorescent tagged Anti Human IgG Abs: bind to IgG Ab.
Enzyme-Linked Immunosorbant Assay "ELISA"
Employs antibodies that have been labeled with detectable enzyme commonly peroxidase.
Labeled antibodies bind to fixed antigen
Binding can be direct (detects Ag) or indirect (detects Ab).
Antigen location is determined using colorimetric assay (msr Change in color: determine conc.
DIRECT ELISA
Looks for specific antigen.
EX: human chorionic gonadotropin (hCG)
in Pregnancy and Rapid Strep A tests. Specimen placed in wells of microtiter plate.
EX 2: and Rapid Strep A tests Wells treated with antibody Ex Ab to S. pyogenes
INDIRECT ELISA
Food allergy tests.
Looks for antibody in patient serum; Human IgG is added.
Wells of plate treated with known antigen
Western blotting technique
Identify presence of Antibodies in a patient's serum.
Proteins of known antigen are separated using a special gel electrophoresis by size before reacting with antibody.
Makes it possible to establish which proteins are
recognized by antibodies.
METHOD:
1. Electrophoresis of proteins of known Ag.
2. Proteins transferred to a filter by blotting.
3. Patient serum washed over the filter.
4. Matching Ab and the Ag bind.
5. Anti-human serum c/enzyme washed over the filter.
6. Reaction with substrate and enzyme produce colored bands of the filter.
USED TO DIAGNOSE HIV AND LYME DISEASE
Fluorescence Activated Cell Sorter (FACS)
Special version of flow cytometry (Similar to a Coulter Counter) counts cells labeled with fluorescent antibodies.
Used to count subsets of T cells: CD4 and CD8 cells especially (Th and Tc); Antibodies are attached to the CD4 and CD8 markers.
Cells with fluorescently labeled markers
are counted; T cell counts important in keeping track of the progression of HIV into AIDS.
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