Practical Applications of Immunology (Ch 18)
Zazzle.com
Practical Applications of Immunology (Ch 18)
behance.net, CDC.gov
Lady Mary Wortley Montague (1689-1762)
“Sacred to the Memory
of
the Right Honourable
Lady Mary Wortley Montague.
Who happily introduc’d from Turkey,
into this Country,
The Salutary Art
Of inoculating the Small Pox.
Convinc’d of its Efficacy
She first tried it with Success
On her own Children.
And then recommended the practice of it
To her fellow Citizens.
Thus by her Example and Advice,
We have soften’d the Virulence
And escap’d the danger of this malignant
Disease…”
1789
Chapter 18, unnumbered figure B, page 510, Reported numbers of measles cases in the United States,
1960–2010. (CDC, 2010)
140
Reported number of cases
120
Vaccine
licensed
500,000
450,000
Reported number of cases
400,000
100
80
60
40
20
350,000
0
2000
300,000
01
02
03
04
05
Year
06
07
08
09
10
250,000
200,000
150,000
100,000
50,000
0
1960
1965
1970
1975
1985
1980
Year
1990
1995
2000
2005
2010
Live Attenuated: Figure 18.1 Influenza viruses are grown in embryonated eggs.
Live Attenuated: Figure 13.7 Inoculation of an embryonated egg.
Shell
Amniotic
cavity
Chlorioallantoic
membrane
Chlorioallantoic
membrane
innoculation
Air
sac
Amniotic
innoculation
Yolk
sac
Shell
membrane
Albumin
Allantoic
cavity
Allantoic
innoculation
Yolk sac
innoculation
Conjugated vaccines: Figure 17.6 T-independent antigen
West Nile DNA vaccine for horses
nature.com
Adjuvants:
Thimerosal FAQ page from CDC
Mumps
Measles
Measles
Rubella / German measles
wikipedia.org
Diphtheria, skin lesion
Diphtheria
wikipedia.org
Polio, un.org
Tetanus
wikipedia.org
braindiseases.org
freeinfosociety.com
Diagnostic Immunology
Figure 18.2.1-2 The Production of Monoclonal Antibodies.
Antigen
1 Mouse injected with antigen. Antibodies
produced.
2 Mouse spleen removed. Contains B cells
that produce antibodies.
Spleen
Figure 18.2.3-4 The Production of Monoclonal Antibodies.
3 Spleen cells mixed with myeloma cells. Some
fuse into hybrid cells.
Suspension of
spleen cells
Spleen cells
Myeloma cells
Cultured myeloma cells
(cancerous B cells)
Suspension of
myeloma cells
Hybrid cell
4 Mixture of cells placed
in selective medium that
allows only hybrid cells
to grow.
Hybrid cells
Myeloma cell
Spleen cell
Figure 18.2.5-6 The Production of Monoclonal Antibodies.
Hybridomas
5 Hybrid cells proliferate into
clones called hybridomas.
The hybridomas are
screened for production of
the desired antibody.
Desired
monoclonal
antibodies
6 The selected hybridomas produce
large quantities of monoclonal
antibodies, for treating and
diagnosing disease.
healthcentral.com
Figure 18.3 A precipitation curve.
Antigen
Antibody
Zone of
antibody
excess
Precipitate
formed
Antibody in precipitate
Zone of
equivalence
Zone of
antigen
excess
Antigen added
Figure 18.4 The precipitin ring test.
Antigens
(soluble)
Zone of equivalence:
visible precipitate
Precipitation
band
Antibodies
Figure 18.5 An agglutination reaction.
IgM
Epitopes
Bacterium
Figure 18.6 Measuring antibody titer with the direct agglutination test.
1:20
1:40
1:80
1:160
1:320
1:640
Control
(a) Each well in this microtiter plate contains,
from left to right, only half the concentration
of serum that is contained in the preceding
well. Each well contains the same
concentration of particulate antigens, in this
instance red blood cells.
Top view
of wells
Enlarged
photo of
wells
(b) In a positive (agglutinated) reaction,
sufficient antibodies are present in the serum
to link the antigens together, forming a mat of
antigen–antibody complexes on the bottom
of the well.
Side view
of wells
Agglutinated
Nonagglutinated
(c) In a negative (nonagglutinated) reaction, not
enough antibodies are present to cause the linking
of antigens. The particulate antigens roll down the
sloping sides of the well, forming a pellet at the
bottom. In this example, the antibody titer is 160
because the well with a 1:160 concentration is the
most dilute concentration that produces a positive
reaction.
Figure 18.7 Reactions in indirect agglutination tests.
Antigen attached
to bead
Latex
bead
Latex
bead
Latex
bead
Latex
bead
Latex
bead
Latex
bead
Latex
bead
Latex
bead
Latex
bead
Latex
bead
Insert Fig 18.7
Latex
bead
Latex
bead
IgM antibody
Reaction in a positive indirect test for
antibodies. When particles (latex beads here)
are coated with antigens, agglutination
indicates the presence of antibodies, such as
the IgM shown here.
Antibody
attached to bead
Latex
bead
Bacterial
antigen
Reaction in a positive indirect test for
antigens. When particles are coated with
monoclonal antibodies, agglutination
indicates the presence of antigens.
Figure 18.11a Fluorescent-antibody (FA) techniques.
Group A
streptococci from
patient’s throat
Fluorescent dye–labeled
antibodies to group A
streptococci
Fluorescent
streptococci
Reactions in a positive direct fluorescent-antibody test
Figure 18.11b Fluorescent-antibody (FA) techniques.
T. pallidum from
laboratory stock
Specific antibodies Antibodies binding Fluorescent dye–labeled
in serum of patient
to T. pallidum
anti-human immune
serum globulin
Fluorescent spirochetes
(will react with
(see Figure 3.6b)
any immunoglobulin)
Reactions in a positive indirect fluorescent-antibody test
Figure 18.14a The ELISA method.
1 Antibody is adsorbed to well.
2 Patient sample is added;
complementary antigen
binds to antibody.
3 Enzyme-linked antibody specific
for test antigen is added and binds
to antigen, forming sandwich.
4 Enzyme's substrate ( ) is added,
and reaction produces a product that
causes a visible color change ( ).
A positive direct ELISA to detect antigens
Figure 18.14b The ELISA method.
1 Antigen is adsorbed to well.
2 Patient serum is added;
complementary antibody
binds to antigen.
3 Enzyme-linked anti-HISG
(see page 518) is added and
binds to bound antibody.
4 Enzyme's substrate ( ) is added,
and reaction produces a product that
causes a visible color change ( ).
A positive indirect ELISA to detect antibodies
Figure 18.13 The use of monoclonal antibodies in a home pregnancy test.
Control
windows
1 Free monoclonal
antibody specific
for hCG, a hormone
produced during
pregnancy.
Test
windows
2 Capture monoclonal
antibody bound to
substrate.
3 Sandwich formed by combination of
capture antibody and free antibody
when hCG is present, creating a
color change.
Not pregnant
Pregnant
http://www.whfreeman.com/catalog/static/w
hf/kuby/content/anm/kb07an01.htm