BACTERIAL TOXINS

William Edward van Heyningen 1950,

1970

The concept that pathogenic bacteria might elicit their harmful

effects in humans and animals by means of poisons elaborated
by them

Bacterial Virulence Factors

 Damage during infection occurs

Direct action of pathogen
Host response to infection
Combination of both

Virulence

Attachm
ent

Colonizat
ion

Invasion

Toxins
and
enzymes

Inhibition
of
phagocyto
sis

Host - parasite Interaction

Eukaryotic
Cell

Pili or
adhesins

Prokaryotic
Cell

Intracellular
Virulent
Bacteria
Recept
or

Prokaryotic and Eukaryotic Interactions

Eukaryotic
Cell

Pili or
adhesins

Prokaryotic
Cell

Intracellular
Virulent
Bacteria
Recept
or

COLONIZATIO

Adherence
blockers

Prokaryotic and Eukaryotic Interactions

Eukaryotic
Cell

Pili or
adhesins

Prokaryotic
Cell

Intracellular
Virulent
Bacteria
Recept
or

COLONIZATIO

Adherence
blockers

INVASION

TOXINS
Edwin Klebs (18341913) for staphylococci,
Robert Koch (18431910) for Vibrio
cholereae,
And Friedrich Loeffler (18521915) for
diphtheria bacillus,
These attempts failed for methodological
reasons that are now understandable.

In 1888, fortune smiled on Emile Roux and

Alexandre Yersin. Since than microbial toxins
have been recognized as the primary virulence
factor(s) for a variety of pathogenic bacteria.

Toxins
Soluble substances that alter the normal
metabolism of host cells with deleterious
effects on the host

(b) Endotoxins are

LIPOPOLYSACCHARID
ES that are a part of
outer membrane of cell
wall of Gram ve
bacteria and are
released when bacteria
die and cell wall breaks
apart

Exotoxin

Endotoxin

Protein

Lipopolysaccharide

Heat labile

Heat stable

Actively secreted by living cells

Integral part of cell wall released only

on disruption of bacterial cell

Highly antigen

Weakly antigenic

Antitoxin is formed against them

Antibody is formed

Converted to toxoid by formaldehyde

Cannot toxoided

High potency

Low potency

Produced mainly by Gram Positive

Bacteria and also by some Gram
negative bacteria like V. cholerae,
ETEC, Clostridium

Gram -ve Organisms only

Toxins Attacking GIT

Staphylococcus
Bacillus cereus
Clostridium perfringens
Clostridium botulinum
Clostridium difficile
E. Coli
Shigella dysenteriae
Vibrio cholera
Vibrio mimicus
Vibrio parahaemolyticus
Pseudomonas aeruginosa
Aeromonas hydrophillia

taphylococcal Enterotoxi
PYROGENIC TOXINS
They cause immunostimulation and nonspecific T-cell
proliferation.
Cystine loop required for proper conformation and which is
probably involved in the EMETIC ACTIVITY

Little is known about Staphylococcal Enterotoxins

may have a direct effect on intestinal epithelium and on the
vagus nerve, causing stimulation of the emetic center and of gut
transit
Highly resistant, Heat Stable

Sudden onset
Nausea
Vomiting
Diarrhea
Generally self limiting
disease

 Activation of Second
Messenger
Escherichia. Coli
Clostridium botulinum
Bacillus cereus
Clostridium perfringes
Vibrio cholerae /cholera toxin

Painless watery diarrhea (Rice water stools)

Dehydration

Nucle
us

Clostridium difficile
Large molecular toxins A &
B
Antibiotic associated colitis with
bloody diarrhea
Pseudomembranous colitis

Cytotoxic
Disrupt the actin cytoskeleton and tight junctions
Fluid accumulation and destruction of the intestinal
epithelium.
Release various inflammatory mediators from intestinal
epithelial

Rounding of the cells, membrane Blebbing,

Apoptosis and death of the target cell.

Aeromonas
Associated with gastrointestinal disease
Chronic diarrhea in adults
Self-limited acute, severe disease in children
resembling shigellosis with blood and
leukocytes in the stool
3% carriage rate

Aerolysins?
Cytotoxin

Pore former

CYTOTONIC
like ETEC and
Cholera Toxin

Vibrio paraheamolyticus
Usually self-limiting and of moderate severity.
Watery diarrhea, Abdominal cramps, Nausea,
Vomiting, and Headache

THERMOSTABLE
DIRECT HEMOLYSIN
(TDH)
Pore - forming toxin, with the pores
estimated at 2 nm in diameter

TDH-RELATED
HEMOLYSIN
(TRH)

Clostridium botulinum Food poisonin

formed toxin in food .

e primary source of botulism was smoked blood sausages.

Dr.Justnus Kerner posited that there
was something in the spoiled
sausages that brought on the
disease- something he called
wurstgift (German for sausage
poison)
Abdominal pain
Nausea
followed by blurring of vision,
diplopia, and difficulty in breathing.
Peaking
Death due to respiratory failure

Clostridium perfringes
Enterotoxin by Type A
Action like CT ,ETEC in diarrhea
8 to 22 hours after ingestion of contaminated food.

lso causes

Necrotising enteritis : Type C

toxin
eck, Germany, where it was called "Darmbrand,"meaning "firebowels".

e formation at plasma membranes of susceptible cells.

endothelial cells and induces vascular necrosis.
Pig-bel : excessive eating of pig Meat
abdominal cramps, shock, and
bloodydiarrhoea.

Bacillus cereus Toxins

Emetic toxin
Cerulide

Enterotoxin

Emesis

Multicomponent protein

I.Period of 0.5-5 hours

Abdominal pain ,watery diarrhea

Preformed in foods

I.Period of 8-16 hours

Stimulates vagus nerve binding

5HT3 receptors

Formed in intestines of host

Lytic toxin, heamolytic,s
hows fluid accumulation on
ligated rabbit ileal loop

Shigella toxin

Toxins effecting Musculoskeletal system

Clostridium. tetani
Clostridium. botulinum
Clostridium . perfringens

Clostridium tetani
Tetanospasmin

Locked jaw

Ophisthotonus

ostridium botulinum

Liberated during growth.

Seven types of toxins (A-G).
Antigenic (light and heavy chain).
Environmental survival. (Inactivated by heat 100C for
20min ).
Lethal dose= 1-2 g .

MW 70000
Relatively Stable
Symmetric descending paralysis
Onset marked by DIPLOPIA Dysphagia Dysarthria
Death by respiratory paralysis

ins affecting the Nervous Syst

ins affecting Skin & Soft Tissu

Exfoliating toxin of Staphylococcus .
Bacillus.anthracis (Cutaneous anthrax).
Alpha toxin of Clostridium.perfringens
etc.

the epidermal layers upon the hydrolysis of Dsg-1.

Exfoliating Toxin of
Staphylococcus

Exfoliative toxins (also known as epidermolytic toxins) are particularly

interesting virulence factors of S. aureus.
Extremely specific serine proteases recognize

DESMOSOMAL
CADHERINS

FEVER, MALAISE, LETHARGY, AND

POOR FEEDING.
Erythematous
Rash

Only the skin, but not the mucosa.

Fragile, fluid-filled
blisters

lpha toxin of Clostridium perfringen

Hemolytic,
Cytotoxic to PLATELETS AND LEUKOCYTES, AND INCREASES
CAPILLARY PERMEABILITY
Requires CALCIUM
Zinc

Ca
2+

Phospholipase
c
Perfringolysin

Damage the
membrane of cells
by attacking the
phopholipids

Haemol
ysis

ARACHIDONIC
ACID CASCADE

Prostaglandins

Thromboxanes

Leukotrienes

PLATELET AGGREGATES OCCLUDES BLOOD VESSELS

Bacillus anthracis

SUPERANTIGENS
Too Much of a Good Thing

Staphylococcal

Enterotoxins
Exfoliative toxins
Toxic-shock syndrome toxin

Same T cell receptor V chain

domains.Today, 18 different SEs
have been described in the
literature

Streptococcal :
o Group a Streptococci pred.
S.pyogenes
Twelve streptococcal pyrogenic exotoxins (SPEs) A, C,
G-M,
Streptococcal superantigen (SSA
Streptococcal mitogenic exotoxin (SMEZ) 1 and 2
Non Group A

o Streptococcus.equi

 Pyrogenic toxin,
potent stimulation of the immune cell system
MHC Class II and T Cell receptor cells,
Concentrations of < 01 pg/ml sufficient to stimulate the T
lymphocytes in an uncontrolled manner resulting in fever, shock
and death.
Pro-inflammatory cytokines,
TNF-a
IL-1b
IL-2
and shock

Toxic Shock Syndrome

Streptococcal toxic shock Syndrome
Kawasaki disease
Acute rheumatic fever
Auto immune diseases

fever

Toxic shock syndrome (TSS)

hypotension, rash, desquamation, fever and major organ
involvement.

Acute rheumatic fever: Smoot et al. showed that antibodies against

SPE-M and SPE-M* were more common in convalescent sera from ARF
patients compared to patients with pharyngitis

Kawasaki disease: Several investigators reported the selective

expansion of T cells bearing the Vb21 TcR, which points towards a SAg
involvement in the disease

Auto immune diseases:

no direct evidence of SAg involvement
SAgs could, under the right conditions, break the tolerance or
suppression of auto- reactive T cell clones and induce a state of
autoimmunity.
Evidence for this hypothesis came from an ANIMAL MODEL of multiple
sclerosis:
a rapid relapse of the disease

Corynebacte
rium
diphthriae
It
is
an
acidic
,globular protein
with a molecular
weight
most
recently estimated
at 62,000 to 63,000

Clinical features
Malaise, Sore throat, Fever
ADHERENT GREY PSEUDO MEMBRANE
Nasal ulcers,
Obstruction of larynx and lower airways, Difficulty in
swallowing
Lead to Myocarditis, Peripheral neuritis, Paralysis of limbs,

Diphtheria

Pertussis toxin

Complex multisubunit toxin with

an AB5 configuration

PT is known to cause most of the

systemic symptoms associated
with pertussis disease, such as the
profound leukocytosis that may be
a predictor of poor outcome in
infants

Adenyl Cyclase
toxin
increased levels of cAMP affect
hormone activity and reduce
phagocytic activity

Pertussis toxin
ADP ribosylation of G proteins
blocks inhibition of adenylate
cyclase in susceptible cells

Foes can be friends too

Exploited to further basic knowledge of cell biology or for medical
purposes.
CHOLERA TOXIN
LABILE- TOXIN OF E.
COLI

B. PERTUSSIS
TOXIN,

Shigella toxin

Adenylate Cyclase
activity
Action of cAMP as
Second messenger

Purge murine (and

potentially human)
bone marrow of
malignant CD77+ B
cells before an
autologous bone
marrow transplant

Diphtheria toxin,
Pseudomonas
exotoxin A, or the
plant toxin ricin

Streptokinase

Cell- killing component of

immunotoxins

Clear blocked arteries in

patients who have heart
attacks

Toxoided: Tetanus,
Diptheria
toxin,pertussis
toxin etc

Vaccines

BOTULINUM TOXIN TYPE A injectable selective muscle-weakening

agent
1970s by Alan Scott, a physician in California, who used it to treat
Disorder
Dysfunction
strabismus patients.
Chronic headache

Tension type headache

Overactive BLADDER

Spastically contraction
of detrusor muscle

LARYNGEAL DYSTONIA

Vocal Cord Dystonia

Temporomandibular
Joint Dislocation

Non relaxing Uretheral

Sphincter

Tourettes Syndrome

Involuntary,rapid,sudde
n movements of
Muscles

Tremors, ticks
Focal Hyperhidrosis

Writers cramps

Excessive sweating

BoNT/ A is also used cosmetically to

reduce deep wrinkles caused by the
contraction of facial muscles.

Detection

Detection of the pathogen

Nonhuman Experimental Data for Enterotoxins

In vivo assays
1. Ligated intestinal loops
2. Perfusion studies
3. Oral inoculation
4. RITARD MODEL :The
reversible ileal tie adult
rabbit diarrheal disease
mode

Tissue culture Assays:

Chinese hamster
ovary (CHO) and Y-1
adrenal cells
Cytotoxicity assay on
HeLa cell line.
. USSING CHAMBERS

DNA probes, PCR

Enzyme Assays:
ELISA
BD GeneOhm Cdiff assay provides a rapid method for the
qualitative detection of the C. difficile toxin B gene (tcdB) in
diarrheal specimens from patients suspected of having CDI.
This test is based on the amplification of the tcdB gene and
the detection of the amplified DNA using fluorogen-labeled
probes

 Pore Formers
Aeromonas. hydrophila
Clostridium .perfringens
Escherichia coli
Listeria. monocytogenes
Staphylococcus. aureus
-toxin Streptococcus pneumoniae
pneumolysin
Streptococcus pyogenes
streptolysin O

 Inhibit protein synthesi

Corynebacterium diphtheriae/
diphtheria toxin
E. Coli
Shigella dysenteriae
Shiga toxins P
Pseudomonas aeruginosa
exotoxin A

 Activation of Second
Messenger
Escherichia. Coli
Bacillus anthracis/edema factor
Bordetella pertussis
Dermonecrotic toxin
Pertussis toxin
Clostridium botulinum
Clostridium difficile
toxin A
toxin B
Vibrio cholerae /cholera toxin