Essentials of

Immunology
Chapters 28 and 29
The Oldest Vertebrate War
Virulence of Microorganisms -vs- Resistance of Host
THEM - Virulence:
Invasiveness
Toxigenicity
US - Resistance:
Acquired or Induced Immunity (resistance):
Humoral Immunity: mediated by antibodies
Cellular Immunity: mediated by cells (T cells)
Natural Resistance (Innate immunity):
Cells: macrophages, etc.
Mechanical barriers: skin & mucous membranes
Chemical factors: intereferons, fatty acids on skin
Microbial factors
2
Innate Immunity (Natural
Resistance) is Non-Specific
Mechanical barriers
Skin: very few microorganisms penetrate
Mucous membranes: mucous traps many airborne bacteria;
coughing/sneezing expel a portion of the microbe-laden mucous
into the atmosphere, the rest are carried into the stomach were
high acidity and digestive enzymes kill many microbes
Found in nasal-pharyngeal region and the upper respiratory tract and
intestinal epithelium [areas open to outside] as well as genourinary
Much more susceptible to penetration than the unbroken skin
Chemical or physical damage of mucous membranes increases the
chance of infection and disease
Tears: flushing of eyes (contains lysozymes)
Respiratory tract: ciliated epithelium lines the walls; cilia beats
upward, push bacteria, mucous and other particulate material
upward to where it is expelled in saliva and nasal secretions
3
Innate Immunity (Natural
Resistance)
Chemical factors
Fatty acids: attack Gram-negative on skin surface
Bile salts: in gall bladder, liver, intestines inhibit Gram-positive
Lysozyme: in tears and saliva
More effective against Gram-positive
Phagocytins: found in leukocyte
Complement: microbial cell membranes and some cancer cells
can lose physical integrity and lyse in presence of complement
Interferon: active against viruses

4
Innate Immunity (Natural
Resistance)
Microbial factors:
Normal flora of the body provides protection by competition with
pathogens
Example: Gram-positive making fatty acids on the skin
If damaged by antibiotics, the individual may get a fungal
infection
Cellular factors:
Phagocytic cells may engulf microorganisms and destroy it
5
Non-Specific Immunity
Phagocytes
On rare occasions, pathogens break through the physical and
chemical defenses one of the first cells that they encounter
are class of cells called phagocytes (cell that eats)
The primary function of phagocytes is to engulf and destroy
pathogens
Phagocytes can also act as antigen-presenting cells (APCs) and
generate a peptide antigen(s) that will activate specific
immune responses
3 main phagocytes: neutrophils (polymorphonuclear
leukocytes), monocytes and macrophages
Neutrophils: actively motile granulocytes containing large
numbers of lysosomes


6
Major Immune Cell Types: 29.4
(a) nucleated cell in
lower left is
neutrophil
segmented nucleus
granular cytoplasm
(a) Monocyte: slightly
above and to right

(b) Circulating
lymphocyte: no
visible cytoplasm
7
Phagocyte
Toxic oxygen compounds
produced in phagocytes to
destroy pathogens
Toxic compounds include
hydrogen peroxide (H
2
O
2
),
superoxide anions (O
2
-),
hydroxyl radicals (OH),
singlet oxygen (
1
O
2
),
hypochlorous acid (HOCl),
and nitric oxide (NO)
8
Phagocytes: 3 Functions
Phagocytosis:
engulfment
Pathogen
Destruction: by the
reactive oxygen
Antigen Processing:
Target cell lysis
Inflammation
9
Antigen Processing
Presentation to T
H
2 cell to activate B cells plasma cells antibodies
10
Destruction of Pathogen by
Complement and Antibodies
11
Overview of Innate System
12
Induced Immunity (Induced
Resistance)
Requires prior contact or exposure to antigen
Humoral Immunity: mediated by antibodies produced by B-
lymphocytes that differentiate into plasma cells that produce
antibody
Cellular Immunity: mediated by cells
Example: cytotoxic T-cells
13
Specific Immune Response
Humoral Immunity: Antibodies
B cells (lymphocytes) plasma cells Antibodies
Cellular Immunity: T-Cells
Precursor T Lymphocytes antigen reactive/activated T-cells
T-Cells:
CD4: T-helper (T
H
1 and T
H
2): helps or induces an immune
response; associated with MHC Class II
CD8: T-cytotoxic cells; associated with MHC class I
Characteristics of specific immunity:
1. Specificity
2. Memory: Capacity to respond more quickly and vigorously
after exposure to an antigen
3. Tolerance: acquired inability to make an immune response to
certain antigens
14
15

16
PAMP Interaction with PRR
PRR receptor on surface of
phagocyte
For fast and effective
interaction with a pathogen
Facilitates recognition of
pathogen associated
molecular patterns (PAMP)
Pattern recognition
receptors (PRR): membrane
bound phagocyte proteins
Interactions activate
phagocyte to ingest and
destroy

17
Overview of Immune Response
Pathogens are targeted and destroyed by 3 immune
mechanisms:
I. Innate immunity results from interactions between
pathogen-associated molecular patterns (PAMPS) found as
cell surface components and pattern recognition receptors
(PRRs) found on phagocytes (membrane bound phagocyte
proteins)
Does not require previous exposure to pathogen
Mediated by phagocytes
PRRs first observed in Drosophila, called Toll receptors
Toll-like receptors on human phagocytes recognize specific
PAMPS
Ex. TLR-4, a PRR on human phagocytes recognizes & responds to
interactions with LPS, a PAMP in outer membrane of Gram-negative,
inducing phagocyte activation and immunity to Gram-negative
18
Overview of Immune Response
Innate immunity continued:
Interaction of a PAMP with the phagocyte PRR triggers a trans-
membrane signal that results in transcription of a number of cell
proteins in phagocyte
This transcriptional activation leads to production of toxic oxygen
compounds that lead to pathogen death
H
2
O
2
; O
2
-
; HOCl; NO; OH
.
;
1
O
2
Theres an increase of O
2
use which called respiratory burst

19
Inhibition of Phagocytosis
Staphylococcus aureus produces carotenoids that neutralize
singlet oxygen
Mycobacterium tuberculosis uses its cell wall glycolipids to
absorb hydroxyl free radicals and superoxide anions
Others produce leukocidins
Pathogen ingested and produces leukocidin that kills phagocyte
the pathogen is released
Dead phagocytes make up much of material of pus
Staphylococcus aureus & Streptococcus pyogenes referred to as
pyogenic (pus-forming) pathogens
Streptococcus pneumoniae produces a bacterial capsule which
prevents adherence of phagocyte to bacterial cell
Streptococcus pyogenes produces M-protein that alters the
surface of the bacterial cell and inhibits phagocytosis

20
Inflammation
Nonspecific reaction to stimuli such as toxins and pathogens
Causes redness (erythema), swelling (edema), pain, and heat
typically localized to site of infection
Molecular mediators of inflammation include proteins called
cytokines and chemokines produced by various immune cells
but especially phagocytes and lymphocytes
Inflammation is typical outcome of an innate or an adaptive
immune response and typically protective
21
Inflammation
An effective response isolates and limits tissue damage
Rapid localization and destruction of pathogen by recruited
neutrophils and macrophages
Sometimes septic shock can occur if reaction is not localized
and spreads through entire circulatory and lymphatic system
Ex: Gram-negative bacteria - even though cleared, LPS interacts
with TLR-4 stimulating production of cytokines that induce
systemic responses that can have serious consequences like
increased vascular permeability leading to massive efflux of fluids
from central vascular tissue: loss of blood pressure, severe edema
in tissues, loss of blood volume

22
Overview of Immune Response
Pathogens are targeted and destroyed by 3 immune
mechanisms:
I. Innate immunity results from interactions between
pathogen-associated molecular patterns (PAMPS) found as
cell surface components and pattern recognition receptors
(PRRs - membrane bound phagocyte proteins) found on
phagocytes
Adaptive immunity: due to antigen specific T cells, resulting in
two distinct effector pathways
II. Antibody mediated immunity (humoral immunity): results
from soluble antigen-specific antibody proteins, products of
antigen stimulated B lymphocytes plasma cells antibody
T
H
2
III. Cell-mediated immunity: mediated by antigen specific T cells
T
C
and T
H
1

23
Origins of Cells of Immune
Response
Lymphoid precursor can generate T and B cells
Myeloid precursor can generate
Monocytes which in turn develop into macrophages or dendritic
cells that are phagocyte cells involved in antigen uptake and
display
Granulocytes which include neutrophils, mast cells, eosinophils,
and basophils
Neutrophils are also phagocytes
The others release their granule contents in response to pathogens,
pathogen products, or damaged host cells
24
Cells of Immune Response
Lymphoid Precursor:
Maturation in thymus: T cell
Maturation in bone marrow or fetal liver: B cell


25
Acquired Immunity: Induced
Acquired immunity is a state of altered responsiveness to a
specific substance induced by prior contact with the foreign
substance
Ex: Contact with Corynebacterium diphtheriae
Anamnestic: has memory therefore repeated contact with
antigen amplifies response
Immunogens: are capable of eliciting an immune response
26
Acquired Immunity: Induced
Characteristics of immunogens:
Usually protein or complex polysaccharides (molecular
complexity) of a microbe, these are on its surface
Nucleic acids, simple polysaccharides, and lipids are typically poor
immunogens because theyre composed of repeating monomers
High molecular weight: > 10,000 Daltons (10kDa)
Appropriate physical form: large, complex macromolecules in
insoluble or aggregate form (example proteins precipitated by
heating) are good immunogens
Insoluble readily taken up by phagocytes, leading to immune
response
Foreign to responder; not part of self (immunological tolerance to
self antigens)
Note: epitopes on antigen are recognized by antibody

27
Antibody
Antibodies are synthesized by plasma cells in humoral
response and are capable of combining with the provoking
antigen
5 classes
IgG, IgM, IgA, IgE, IgD: different heavy chains
Ex: IgG ( heavy chains); IgM ( chains)
Fab & Fc portions
Also subclasses IgG 1 2 3 4 heavy chains

28
More Terms
Immunogen: substance capable of eliciting an immune
response
An antigen may or may not be an immunogen
Antigens: substances that react with antibodies or TCRs (T cell
receptors)
May not have actually induced/activated them
Hapten: good example of antigen but not immunogen
Low molecular weight substance which can not act as an
immunogen by itself
When attached to high molecular weight materials, an antibody
response is made to the carrier (the high molecular weight
material), as well as to the hapten complex is immunogenic
but not hapten alone
Next time hapten enters it acts as antigen and can react
immediately with those antibodies made to its epitope(s)


29
Epitope
Antibody or TCR does not interact with the antigenic
macromolecule as a whole, but with distinct portions of the
molecule called the epitope (formerly called antigenic
determinative)
Antigens (most are immunogens) are complex; often >1 epitope
Antibody specificity is sensitive enough to distinguish between
two very similar epitopes
Ex: glucose from galactose which only differ in the orientation of
a -OH group

30
MHC Proteins:
Antigen Presentation
MHC: Major Histocompatibility Complex (proteins)
Function as antigen presenting molecules and interact with
both antigen and later TCR (T Cell Receptor)
T cell cannot recognize foreign antigen unless it is presented in
context of an MHC protein
MHC: think of it as a platform that brings degraded antigens and
presents them to the TCR
MHC in humans have differences (polymorphism) which leads
to an issue during organ transplant
Two classes:
1) Class I: found on surface of all nucleated cells
2) Class II: found only on surface of B lymphocytes, macrophages,
and dendritic cells, all dedicated APCs (antigen presenting cells)

31
MHC Class I
MHC I proteins are made and assembled
in the ER
Chaperon proteins stabilize MHC I until
antigen is bound
Protein antigens manufactured within
the cell (such as from viruses or tumors)
are degraded by the proteasome in the
cytoplasm, transported across the ER
through a pore formed by TAP proteins
They then bind to MHC Class I,
transported to cell surface to bind with
TCR (CD8 coreceptor makes binding
stronger)
T
C
cell then releases cytokines and
cytotoxins to kill the target cell



32
MHC Class II
MHC II proteins are produced in the ER
and assembled with a blocking protein (Ii,
invariant chain, which prevents MHC II
from complexing with peptides found in
the ER)
Lysosomes containing MHC II fuse with
phagosomes forming phagolysozome
where the proteins from outside the cell
[brought in by endocytosis] are digested (li
is also digested)
MHC Class II then binds to digested foreign
proteins and the complex is transported
out of cell to bind to TCR and CD4
coreceptor on T
H
cell T
H
release
cytokines which convert the activated T
cell to T
H
1 or T
H
2
Only APCs can be targets for T
H
cells recognizing
the peptide-MHC II complex


33
Comparison MHC Class I & II
34






Note: Peptide bound by both MHC I and TCR Proteins
35
T Cells: Cytotoxic & Helper
T-Cytotoxic Cells (T
C
): destroy cells that
display antigens embedded in MHC class I
molecules
Granules migrate to contact site and
content is released
Uses perforin (forms pore to deliver toxic
enzymes) and granzymes (proteins) that
enter through pore and cause apoptosis
Intracellular pathogens (such as viruses)
hide inside host cells, where they are
protected from antibodies
Best killed when host cell sacrificed
Class I MHC found on all nucleated cells
and thus any infected cell can activate T
C

which kills infected cell
36
37
T
H
(T
Helper
) Cells: T
H
1 and T
H
2
Macrophages: central role as APCs which bind,
process and present antigen to T
H
cells
T-inflammatory (T
H
1) cells are activated by
antigens presented on macrophages in the
context of MHC Class II protein increased
macrophage activity and inflammation
T
H
1 cells then produce cytokines that stimulate
macrophages to take up and kill certain foreign
cells by themselves increased phagocytosis
Activated macrophages can kill intracellular
bacteria that would normally multiply
T
H
1 also secrete IL-2 cytokine which is a 2
nd
signal
needed for activation of T
C
cells
1
st
signal was the interaction with MHC Class I
peptides on APC
Thus, T
H
1 involved in cellular immunity

38
T
H
2: Promotes Antibody
Production
T
H
2 cells play a key role in B cell
activation and antibody production
B cell coated with antibody that acts
as antigen receptors
Antigen endocytosed and degraded
are presented by MHC Class II
protein to T
H
2
Activated B cells are stimulated by
IL-4 and IL-5 which directs
differentiation into antibody
producing plasma cells
Other cytokines direct
differentiation into memory cells
39
40
Summary
Intracellular pathogens (such as viruses) generally activate cell
mediated immunity by stimulating cytotoxic T cells (T
C
)
Extracellular pathogens tend to activate humoral immunity
(T
H
2)
T
H
2 activates B cells to make antibody
T
H
1 activates macrophages to enhance killing of engulfed
pathogen promotes inflammation
Also further stimulates T
C
cells
41
Antibodies (Immunoglobulins)
Antibodies (Ab)/immunoglobulins (Ig) are protein molecules that are
able to combine with antigenic determinates/epitopes
Found in the serum and in other body fluids such as gastric
secretions and milk
Serum containing antigen-specific antibody is often called antiserum
Comprised of 5 major classes on the basis of their physical,
chemical, and immunological properties
IgG, IgA, IgM, IgD and IgE
About 80% of the serum antibodies are IgG proteins
Do not directly kill the pathogen
Mark it for destruction (opsonization)
IgM or IgG may attract the complement proteins
IgA blocks pathogen interaction with host cell
42
Opsonization
43
Immunoglobulins
All immunoglobulin classes have
variable domains
V
H
= variable heavy
V
L
= variable light
Both bind to antigens
All immunoglobulin classes have
constant domains
It is the difference in the amino
acid sequence of the constant
domain that defines the class of
immunoglobulin molecule
Gamma (), alpha (), mu (),
delta () or epilison ()
F
C
crystallizes , Fab does not
44
Papain Reaction with IgG
Papain: nonspecific thiol-endopeptidase
Enzymatically cleaves IgG just above the hinge region to create 2
separate Fab fragments and 1 Fc
Fab fragments bind antigen; antibody-bivalent
Purification scheme shown below
45
Molecular weight/ Serum Antigen- Properties Distrubution
Class/Chain formula mg/mL binding sites
IgG (gamma) 150,000 2(H+L) 13.5 2
Major circulating
Ab; 4 subclasses;
activate
complement
Extracelluar
fluid;blood & lymph;
crosses placenta
IgM (mu)
970,000 (pentamer);
5[2(H+L)] + J;
175,000 (monomer);
2 (H+L)
1.5
0
10 (pentamer),
2 (monomer)
1st Ab to appear
after immunization;
strong complement
activator
Blood & lymph;
monomer is B cell-
surface receptor
IgA (alpha)
150,000 (monomer);
2(H+L);
385,000 (secreted
dimer);
2[2(H+L)+J+SC
3.5
0.05
2 (monomer),
4 (secreted)
Important
circulating Ab: -
Major secretory
antibody
Secretions (saliva,
colostrum, cellular &
blood fluids);
monomer in blood
and dimer in
secretions
IgD (delta) 180,000, 2(H+L) 0.03 2 Minor circulating Ab
Blood & lymph; B
lymphocyte surfaces
IgE (epsilon) 190,000; 2(H+L) 0.00005 2
Involved in allergic
reactions; C
Helper4 contains
mast cell binding
fragment
Blood & lymph; binds
to mast cell surfaces
Table 22.2 Properties of human Immunoglobulins
46
IgG Antibody
Most common of the circulating
antibodies
Composed of 4 polypeptide
chains
Interchain disulfide bridges (S-S)
connect the individual chains
A functional IgG molecule
consists of two antigen binding
sites
IgG is therefore bivalent and can
bind two identical epitopes
Crosses the placental barrier
Subclasses: gamma 1, gamma 2,
gamma 3, gamma 4
47
IgM Antibody
Usually found as an aggregate of
five immunoglobulin molecules
attached by at least one J
(joining) chain
Every heavy chain of IgM contains
a fourth constant domain (C
H
4)
1
st
class of immunoglobulin made
in a typical immune response to a
bacterial infection
10 binding sites
Low affinity but high avidity
Monomers on B cell surface

48
IgA Antibody: Secretions
Present in the serum in the monomeric form, but in secretions it is
a dimer
Colostrum (breast milk), mucosal secretions of gastrointestinal, tears,
respiratory and genitourinary tracts
IgA dimer linked by disulfide bridges to J chain protein
Secretory piece wrapped around IgA dimer during secretion.
Total amount produced is twice the amount of IgG in serum
49
IgE Antibody
Found in the serum in very small amounts
Antibody that binds to eosinophils, arming these granulocytes
to target eukaryotic parasites
Ex: schistosomes and other worms
Mediates immediate-type hypersensitivities (allergies)
Very mild or life threatening (anaphylaxis)
Like IgM, IgE has a fourth constant domain (C
H
4)
Constant region functions to bind IgE to mast cells surfaces
Mast cell degranulation causes release of mediators such as
histamine and serotonin
Leading to dilation of blood vessels and contraction of smooth
muscle

50
Immediate Hypersensitivity:
Type I
51
Types of Hypersensitivity
52
IgD Antibody
Present in the serum in low concentrations
Has no know function
Abundant on the surfaces of B cells and plays a role along with
monomeric IgM in binding antigen to B cells
Especially abundant on memory cells
"Immunoglobulin D enhances immune surveillance by
activating antimicrobial, proinflammatory and B cell-
stimulating programs in basophils
2009. Nat Immunol
Insights into the function of IgD.
Dev Comp Immunol. 2011 Dec;35(12):1309-16. doi:
10.1016/j.dci.2011.03.002. Epub 2011 Mar 22.
Edholm ES, Bengten E, Wilson M.

53
Summary of Antibody
Production
I. Antigens are spread via lymphatic and blood circulatory
systems to neighboring secondary lymphoid organs [lymph
nodes, spleen, or mucosal-associated lympoid tissue
(MALT)]
II. Intravenously injected antigen travels via the blood to the
spleen, where antibodies are formed
III. Subcutaneously, intradermally, topically, or intraperitoneally
introduced antigens are carried by lymphatic system to the
nearest lymph node
IV. Antigens introduced to mucosal surfaces (such as the
mouth) are delivered to the GALT lining the intestinal tract
resulting in antigen-specific IgA antibody production in the
gut
54
Lymphatic System
Lymphatic system is part of the immune system,
made up of a network of conduits that carry a
clear fluid called lymph
It includes the lymphoid tissue and lymphatic
vessels through which the lymph travels in a one-
way system
Lymph flows only towards the heart
Lymphoid tissue is found in many organs,
particularly the lymph nodes, and in the
lymphoid follicles associated with the digestive
system such as the tonsils
The system also includes all the structures
dedicated to the circulation and production of
lymphocytes, which includes the spleen, thymus,
bone marrow and the lymphoid tissue associated
with the digestive system
Movement of leukocytes from blood
interstitial space lymphatic vessel blood is
known as extravasation


55
Lymphatic System
56
Primary lymphoid organs:
1. Thymus
2. Bone marrow
Antibody Production:
Primary Response
Following initial antigen introduction, each antigen-stimulated B cell
(via T cell activation) multiplies and differentiates to form both
antibody-secreting plasma cells and memory cells
Plasma cells are short-lived (< 1 week) but produce large amounts of
IgM antibody in this primary antibody response
Following a latent period, specific antibodies show up in the blood,
followed by a gradual increase in antibody titer (quantity), and then
a slow fall in the primary response
57
Antibody Production:
Secondary Response
Memory cells generated by the initial exposure to antigen may
live for several years
Upon re-exposure to the immunizing agent, memory cells
need no T cell activation quickly transform to plasma cells
and begin producing IgG
The antibody titer rises rapidly to a level 10-100x greater than
the titer following the initial exposure
The rise in antibody titer is referred to as the secondary antibody
response (anamnestic)
The secondary response is characterized by a switch from IgM
to IgG production (class switching)
Over time, titer slowly decreases but a later exposure can
cause another secondary response
Basis for booster shots
58
59
60
Recognize Two Divisions of
Immune Response
From Nature and Experiments
1. Humoral/antibodies Immunity:
B lymphocytes differentiate into plasma cells and produce
antibody
B lymphocytes mature in bone marrow/fetal liver
Chickens - remove the bursa and they fail to produce antibodies and
become susceptible to bacterial or extrinsic invaders
Humans - Burtons Agammaglobulinaemia: children fail to produce
antibody. Humans do not have a bursa, but instead have GALT (gut
associated lymphoid tissue) and MALT (mucosal cells that line
external surfaces)
In both of the above examples, cellular immunity remains essentially
intact and they are more susceptible to bacterial infections rather
than viral or intrinsic
61
Recognize Two Divisions of
Immune Response
From Nature and Experiments
2. Cellular Immunity:
Mediated by cells
Involve lymphocyte processing by thymus and activated T cells
Where antibody mediated (humoral) recognizes substances that
are outside host cells (extrinsic), cell-mediated immune response
is more effective in recognizing modified host cells (intrinsic)
It is important in controlling infections in which pathogens can
reproduce within living cells
Ex: viruses, some bacteria like Rickettsia and Chlamydia, and some
parasitic protozoans like typanosomes
By T
C
and NK cells
Also important in surveillance/destruction of malignant cells
Phagocytosis (destruction of extrinsic) and antigen presentation
62
Cellular Immunity: Natural
Killer (NK) Cells
A fourth cell type (NK cells) are involved in innate immunity
1) T
H
1: CD4; Class II MHC; 2
nd
signal activating cytotoxic T
cells (cytokines: IFN-, IL-2, TNF-) inflammation
increased phagocytosis
2) T
H
2: CD4; Class II MHC; B-cell helper (cytokines: IL-4, IL-5, IL-
6)
3) T
C
: CD8; Class I MHC; killing virus-infected and cancer cells
(cytokines: IFN-, TNF)
4) NK: class of lymphoid cells that instead of killing microbes
destroys host cells that harbor microbes or have been
transformed into cancer cells
Recognize changes in cell surface proteins of compromised cells
and then degranulate to release chemicals that kill them
Innate: reacts early, non-specific
63
NK Cells
NK cells kill tumor cells, virus-infected cells, bacteria, fungi,
and parasites
Non-specific
Involved 2 ways in innate immune response:
1) Target-cell killing mediated by initiating apoptosis in target cell
Release perforin and granzyme
Perforin polymerize in target cell membrane forming pore used by
granzyme to enter and stimulate apoptosis
2) Production of cytokines including TNF and granulocyte-
macrophage colony stimulating factor
Does not use TLRs (toll-like receptors)
Protective mechanism that takes place in the interval before
the more powerful adaptive immune response is active
Number of cells does not increase or have memory
Recognize a decrease in MHC class I on the surface of cells
64
Recognize Two Divisions of
Immune Response: From
Nature and Experiments
Cellular (continued):
Cell mediated immune responses include:
Surveillance/destruction of malignant cells
Delayed hypersensitivity in response to intracellular bacteria like
Mycobacterium tuberculosis and fungal infections such as
Histoplasma capsulatum
Cytotoxic T lymphocyte (T
C
) responds to virally infected cells
Response to tumor cells and tissue grafts by natural killer (NK) cells


65
Recognize Two Divisions of
Immune Response: From
Nature and Experiments
Cellular (continued): Experiments and nature
Thymectimized mice:
More susceptible to viral infections or intrinsic parasites like M.
tuberculosis
Increased rate of tumor production
Failure to reject transplanted tissue
Di Georges Syndrome (humans):
Infants lacking a functioning thymus gland (equivalent to
thymectimized mice above)
10,000 times greater chance of malignancy but not susceptible to
bacterial extrinsic infection
66
A Note
Cellular immunity (T
C
mediated) and innate NK cells provide
an immunosurveillance mechanism to rid the body of old,
dead, damaged, or mutated (malignant) cells
This mechanism is impaired in individuals on
immunosupressive drugs, of old age or if chemicals are
present (pollutants, drugs, radiation, UV, cigarette smoking,
insecticides, etc)

67
68
Natural vs Artificial & Active vs
Passive Acquired Immunity
I. Active Immunity: antibodies (Ab) made or T cells activated
in the individual upon contact with antigen (Ag)
Longer lasting memory cells
1. Artificial: individual purposely exposed to controlled dose of
harmless Ag to induce artificial active immunity
Process known as vaccination
2. Natural: animals normally develop natural active immunity by
acquiring a natural infection that initiates the adaptive immune
response
II. Passive: cells or Ab from an immune individual are
transferred to a nonimmune individual to prevent or help
cure a disease

69
Outcome of Host-Pathogen
Interaction
Vaccination: inoculation of host with inactive or weakened
pathogens or pathogen products to stimulate immunity
Attenuated strains: immunocompromised individuals?
Chemically or physically inactivated strains (formaldehyde treated
polio virus: Salk polio vaccine)
Products of pathogens: some genetically engineered and
produced in large quantities
Example: Tetanus toxoid - inactivated exotoxin
Example: Anthrax - protective factor
New: recombinant vector vaccine
Example: Vaccinia - rabies vaccine used in animals (V-RG)
New: DNA vaccines
Bacterial plasmid with cloned DNA injected intramuscularly into host
animal T
C
cells, T
H
1 cells, Ab made to protein encoded by cloned
DNA
70
Attenuation
Attenuated strains have lost virulence
Often they retain immunogenicity, therefore they may be used
for production of vaccines
Especially viral vaccines: measles, mumps
Laboratory cultivation typically results in a decrease in
virulence of pathogens or even a complete loss
Danger for immunocompromised individuals even though live
cells or viruses are generally more effective than immunization
with dead or inactivated

71
72
73
Examples of Passive Immunity
Passive: Ab or cells activated in one individual are transferred to
another
Short lived compared to active
1. Artificial:
Tetanus antiserum is given to passively immunize an individual
suspected of being exposed to Clostridium tetani due to acute injury in
car accident
Contrast with vaccination (a prophylactic measure) with tetanus toxoid
that actively immunizes individual for future encounter with Clostridium
tetani exotoxin
Pooled human gamma globulin given to patients exposed to hepatitis A
2. Natural: transfer occurs as part of a natural process
Placental transfer of IgG from mother to fetus
IgA transferred from mother to baby in breast milk
74
75
Antimicrobial Therapy: In Vivo
Antimicrobial agent is a natural or synthetic chemical that kills or
inhibits growth of microorganisms in vitro and in vivo
Chemotherapeutic agent are those used to treat microbial
disease and now also to prevent proliferation of malignant cells
An in vivo agent includes antibiotics: (Greek for against life)
Compounds produced by one species of microbe that can kill or
inhibit growth of other microbes (original definition)
Comprise the vast majority of chemotherapeutic agents
Today the term is commonly used for synthetic and semisynthetic
agents as well as true antibiotics
Synthetic: like sulfonamides
Semisynthetic: different penicillin derivatives
76
Hospital Acquired: Nosocomial
Infections
Many hospital patients with noninfectious diseases (cancer
and heart disease) acquire microbial infections that produce
disease because they are compromised
Such health-care associated infections are called nosocomial
infections
Procedures like catheterization, hypodermic injection, spinal
puncture, biopsy and surgery unintentionally introduce
microbes into patient
Points out the need to control microbes
Nosocomial are often antibiotic resistant
77
Control of Microorganisms
Physical Agents:
Most widely used is heat
Chemical Agents:
Called antimicrobial agents
Kill or Inhibit Growth:
Sterilization: treatment that frees the treated object of all living
organisms, including viruses
Death: defined as irreversible loss of ability to reproduce when
inoculated into an appropriate medium or host
78
Death as a Function of
Temperature
Death from heating is
an exponential function
Occurs more rapidly as
the temperature rises
Effectiveness is
measured by the time
required to decrease
viability by 10x
Maximum temperature
Moist heat is better
than dry heat
79
Autoclave
An autoclave permits application of steam heat under
pressure at temperatures above the boiling point of water,
resulting in the killing of endospores
At 15 lbs per sq inch above normal atmospheric pressure
water boils at 121C
At that temperature for 15 minutes, endospores are killed if
volume of liquid not too large
Ex. it takes 30 min for 2L flask
High sugar and fat concentration in a media increase the
resistance of an organism to heat treatment
High salt concentration may increase or decrease the
organisms resistance to heat
80
81
Pasteurization
Pasteurization does not sterilize liquids but reduces microbial
load, killing most pathogens and inhibiting the growth of
spoilage microorganisms
Originally: 30 min at 62C
Now: Flash Pasteurization: 72C for 15 sec.

82

Examples of Other Methods
Ultraviolet Light:
Leads to the production of thymine dimers
Most effective at 260 nm
Ionizing Radiation:
Higher energy and shorter wavelength
Membrane Filter Sterilization:
Youve done it in lab!
83
Chemical Control:
Antimicrobial Agents
Antimicrobial agent is a natural or synthetic chemical that kills
or inhibits growth of microorganisms
Kills: -cidal (viricidal, fugicidal, bactericidal)
If kills by lysis: bacteriolytic
Inhibits growth: -static (bacteristatic)
Two classes: In vitro and In vivo
In vitro:
Disinfectants: chemicals that kill microorganisms but not necessarily
spores
Sanitizers: reduce to safe levels
Antiseptics or germicides: kill or inhibit growth but nontoxic enough
to be applied to human tissue
84

85
Bacteriostatic inhibits protein
synthesis, if agent concentration
decreases the cell can grow again
Bacteriocidal kills the cell but does
not destroy the cells
Bacteriolytic kills the cell but does
destroy the cell, leading to release of
cellular content
Methods to Evaluate
Agar Zone Diffusion:
Zones of inhibition
Tube Dilution Test:
MIC: minimum inhibitory concentration (learned in lab)
Vary concentration of agent determines minimum required to
inhibit growth; not turbid
Bactericidal Tests: transfer to medium without agent and test
for growth
Phenol Coefficient Test: involves transfer to fresh medium
Greatest dilution that kills microorganism at 10 minutes but not 5 to
greatest dilution of phenol that kills microorganism in 10 but not 5
minutes



86
87
Kirby-Bauer Disk Susceptibility
Test
After incubating plates, the
diameter of zone of inhibition is
measured and results compared
with a table listing whether a zone
size is wide enough to be clinically
useful
Larger zone of inhibition does not
always mean more effective
May be different solubility
In vivo versus in vitro
88
Tube Dilution Technique
MIC (Minimum Inhibitory Concentration): lowest
concentration of the agent that completely inhibits the growth
of test organism
Bacteristatic Test: only growth inhibition

89
Antimicrobial Agents: In Vivo
Antibiotics: chemical substance produced by one
microorganism that kills or inhibits growth of another
Ex: Penicillin G
Effective against Gram-positive, are Gram-negative impermeable
Inhibit cell wall synthesis
Synthetic Antimicrobial Agents: created in hopes of finding a
magic bullet
Ex: sulfanilamide
Analog of p-aminobenzoic acid, a nucleic acid precursor
Inhibits nucleic acid synthesis by blocking the synthesis of folic acid
We obtain folic acid from our diet
Semisynthetic: other penicillin with modified R group


90
Ehrlich: Selective Toxicity:
Ability to inhibit or kill
pathogen without adversely
affecting host
Ehrlich searched for the magic
bullet
Tested large numbers of dyes
that stained bacterial cells
Discovered first effective
antimicrobial agent, Salvarsan,
arsenical agent effect in
treatment of syphilis
91
Modern Era: Discovery of
Penicillin
Discovery: a great example of
Serendipity in Science
Forgotten until Florey rediscovered
his work just before WWII
Originally recognized by Fleming in 1929
Photo of the plate with mold
contamination
92
93
94
Mechanism of Action
Transpeptidases bind to penicillin and then cannot catalyze
crosslinking reactio
Cell wall synthesis continues but it cannot be crosslinked and
cannot maintain strength
Pen-transpeptidase complex stimulates autolysins that digest the
wall lysis
Known as PBP
Cephalosporins:
Beta lactam ring and a six membered
additional ring instead of the five ring
thiazolidine ring
Semisynthetic and resistant to beta
lactamases
Broader spectrum than penicillin

95
Next Discovery: Domagk 1935
Administered a dose of a red dye to his 6 year old daughter
who had a Streptococcal infection that had spread to her
lymph nodes under arm
It became so severe that the arm needed amputation
She recovered
1928: dye Prontosil did not produce a zone of inhibition on
inoculated plate
When tested in a mouse, the prontosil was very effective at
preventing infection
Prontosil sulfanilamide
Great against Gram-positive cocci
First commercially available antibiotic

96
Growth Factor Analogs: Sulfa
Drugs
97
Mechanism of Action: Sulfa
Drugs
Microbe makes folic acid from precursor para-aminobenzoic acid
(PABA), a vitamin necessary for nucleic acid synthesis
Sulfanilamide resembles PABA in structure, therefore competitive
inhibitor of enzyme that makes folic acid in bacteria
Folic acid is not synthesized by man and instead is obtained in diet
Bacteria do not transport folic acid and therefore sulfa drugs can
inhibit bacterial growth in humans without affecting human host
Today we have some resistant bacterial strains that have gained ability
to transport folic acid
98
Drug Targets
99
Antimicrobial Spectrum of
Activity
Spectrum: range of microbes that a given drug affects
Penicillin G: narrow spectrum (only Gram-positive)
Ampicillin: broad spectrum penicillin (inhibits Gram-negative also)
Vancomycin: narrow spectrum


100
Ideal Properties
1. Selective toxicity
Toxic to microbe but not harmful to man and or higher animals
Some not selectively toxic:
Tetracyclines: taken before adult teeth come in cause black teeth
Chlormaphenicol: implicated in aplastic anemia in children
2. Diffuses to site of infection
3. Very few toxic side effects
4. Allergic reactions not common
5. Broad spectrum
101
Antivirals
102
Antifungal Drugs
Fungi are Eukarya, therefore it is difficult to make drugs
Much of their machinery is the same as in animals and
humans
Drugs that affect a metabolic pathway often affect a
correlating pathway in the host cell
This makes the drug toxic

103
104
105
106