Bacterial Toxins

Endotoxin: cell associated

Exotoxin: secreted from the cell
Lipopolysaccharide (LPS) example of an endotoxin
LPS coats the outer membrane of gram negative bacteria
LPS can be shed from the OM of gram negative bacteria in OM derived
vesicles or OM fragments
LPS is composed of lipids and sugars
Three components are O antigen, core, and Lipid A

Lipid A is the toxic component of endotoxin

The O antigen is not a protein and is unique to each bacterial specie
Heat does not easily inactivate the function of endotoxin compared to
exotoxin
The receptor for LPS in humans is the toll like receptor 4 (TLR4)
TLR4 can be found on variety of cells including macrophages and
endothelial cells
LPS-TLR4 binding induces a variety of responses depending on the cell
(pro-inflammatory cytokine release causing endotoxic shock)
Sepsis: an example of endotoxic shock
Over 750,000 cases per year in the United States of which die of shock
Can be caused by all kinds of microbes
Related to mass replication of microbes in blood
Endotoxic shock is just one type of septic shock
Symptoms: fever, hypotension, inflammation, increased antibody production,
vasodilation, organ failure, intravascular coagulation
Intravascular coagulation is bleeding into tissue causes blood clots to form
which can damage tissue due to blockage of normal blood flow
Then overtime clotting proteins in the blood are consumed increasing the
risk of serious bleeding from even minor injuries

An example is Neisseria meningitidis septicemia which causes disseminated

intravascular coagulation

Exotoxins
Secreted
Cytotoxins: cause cell damage
Pore forming toxins: causes cell lysis
Exoenzymes: alter normal cell function
Superantigens: cause release of vast amounts of cytokine and nonspecific
activation of T cells
Many others
Key point: The microbe that releases the exotoxin can be distal from the site of
the exotoxin action because the exotoxins are secreted and circulated in the body
Properties of Exotoxins
Usually proteins and usually enzymes
Have highly specific targets
Secreted: can affect tissues located away from site of bacterial colonization
Many exotoxins are AB toxins
Antibodies can be used to counteract exotoxins
Neutralizing the toxins enzymatic activity
Prevent receptor recognition and uptake of exotoxin into the cell by binding to
the toxin
AB toxins
Exotoxins are typically AB toxins though they can also be directly injected into
cells by a type III secretion system
The A (active) subunit is enzymatically active and is responsible for the toxic
activity
The B (binding) subunit mediates binding to the receptor on a target cell
The B subunit is what determines the cell and tissue specificity of an exotoxin
The AB toxin is brought into the cell by receptor mediated endocytosis
Once in the cytosol the endosome acidifies which causes the release of the A
subunit
The A subunit exits the endosome and carries out its toxic function

 Responsible for knowing three exotoxins

Diphtheria toxin (AB toxin)
Has 1 A subunit and 1 B subunit
Cholera toxin (A+5B toxin)
Has 1 A subunit and 5 B subunits
Anthrax toxin (2A+B toxin)
Has 2 A subunits and 1 B subunit
Holo-toxin: any toxin whose structure is composed of several subunits
DPT (also DTaP) vaccine
DPT vaccine given to protect against three diseases: diphtheria, pertussis, and
tetanus
Diphtheria and tetanus are both mediated by only one toxin
Pertussis is mediated by more than one toxin as well as other factors
Diphtheria background
Ancient disease that has been present throughout history
Diphtheria infections predominant in young children
From 1735-1740, 40% of children under the age of 10 died in New England
In 1920s US, 100-200k cases per year and 13-15k deaths per year
Diphtheria is now rare due to mass vaccination
Case fatality is 5-10% down from 50% which was the rate before the advent of
antitoxins and antibiotics
Diphtheria clinical information
Transmission: direct contact with infected people or their bodily fluids
Classic diphtheria is caused by Corynebacterium diphtheriae
Colonization of the pharynx by C. diphtheriae results in pharyngitis which is the
inflammation of pharynx resulting in the characteristic bull neck
Diphtheria can also cause cervical edema
In the pharynx, a pseudomembrane forms induced by diphtheria toxin (DT)
Pseudomembrane is bluish-grey in color
Pseudomembrane is composed of fibrin, bacteria, inflammatory cells, and cell
debris
The pseudomembrane is responsible for respiratory obstruction which can
cause death
DT spreads systematically in the body affecting multiple organ systems
Myocarditis (inflammation and damage of heart muscle) in heart
Paralysis (spread to central nervous system)
There is also a cutaneous form of diphtheria which colonizes nasal cavity, larynx,
or less commonly the skin
How to diagnose diphtheria
Look for symptoms such as bull neck, cervical edema, pseudomembrane
(difficulty in breathing)
Take a swab of pharynx to culture bacteria and test for presence of diphtheria
toxin

 How to treat diphtheria

Treat with antibiotics
Erythromycin
Penicillin
Treat with antitoxins
Antitoxin is obtained from the serum of horses immunized with diphtheria
toxoid
Toxoid: a toxin whose toxicity has been inactivated/suppressed chemically
(formalin) or by heat, but still is immunogenic (can elicit an immune
response)
For best clinical outcome, antitoxin should be given ASAP
Corynebacterium diphtheriae (classical causative agent of diphtheria)
Morphology: Gram + rods; distinct club-shaped morphology is indicative
Non-motile
Does not form endospores
Still need to distinguish from normal flora coryneform bacteria that do not cause
diphtheria
Diphtheria toxin is encoded for by the tox gene on bacteriophage genome
This bacteriophage is corynephage
This implies that Corynebacterium that have not been lysogenized with
corynephage will not cause diphtheria
If nontoxigenic C. diphtheriae (i.e. not phage-infected) colonizes the pharynx
then instead of diphtheria, the patient develops pharyngitis i.e. sore throat
Corynephage may lysogenize other members of the genus corynbacterium
which could then go on to cause a diphtheria like disease
ADP-Ribosylating toxins
ADP-Ribosylating toxins are ADP ribosyl transferase enzymes
Depending on the toxin, the host cellular target can vary