Kuliah Mikrobiologi I

2005

Bacterial Pathogenesis
Caroline Tan Sardjono
Importance in understanding
bacterial pathogenesis
• Diagnosis & therapy
• Design for high efficacy
therapy or vaccine
Bacterial Pathogenesis
• Normal flora: microorganisms that are usually
found associated with healthy body tissue
• Parasite: an organism that grow in or on a host
• Pathogen: parasite that does harm to a host
• Pathogenicity: the ability of a parasite to inflict
damage on the host
• Virulence: the degree of pathogenicity produced
by a pathogen
• Infection: growth of organisms in the host
• Disease: injury to the host that impairs host
function
Bacterial Pathogenesis
Definition of an infection
• Organism has entered body (with or without
disease)

Disease
• Inadequate immunity
• Virulence factors
• Infectious dose
Host defence mechanisms
 Humans are continuously exposed to microbes
but it is relatively rare for them to cause disease

Defences include:
- physical barriers
- mechanical processes
- specialised immune cells
- secretory immunity
- production of noxious compounds
- normal flora
1. Physical barriers
• Skin:
- acidic pH,
- contains lactic acid, fatty acids (in sweat)
• Filtration of inhaled air
• Tears: lysozyme
• Acidity: stomach, vagina
2. Mechanical processes
• Flushing: flow of fluid over mucosal surfaces
removes bacteria that are not specifically
attached
- urine flow in urogenital tract
- gut contents in GI tract
- tears over cornea
• Mucocilliary action: in respiratory tract
- surface covered in layer of mucous prevents
bacterial attachment
- ciliated epithelia push mucous upwards
towards the throat
3. Specialised immune cells
• Phagocytic cells part of reticuloendothelial system
- lung  alveolar macrophages
- peridontal tissue  gingival macrophages
- kidney  mesangial cells
- liver  Kupffer cells

• Phagocytic cells
 ingest bacteria  phagosome
 fusion with lysosome  bacteria lysed
Cytoplasm

Phagosome
formed

Lysosome

Phago-lysosome fusion
& bacteria lysis
4. Secretory immunity
• Secretory immunoglobulin A, sIgA:
- inhibits bacterial attachment to mucosal epithelia
- agglutinates bacteria
- binds (inactivates) toxins
- present in colostrum so protects suckling infants

• Does not activate complement or opsonise

bacteria so relatively unimportant in systemic
infection

• Major role in mucosal immunity, prevents

colonisation and infection
5. Production of noxious compounds
• Lysozyme (in saliva, tears, other mucosal fluids)
 cleaves peptidoglycans, disrupts bacterial walls
• Lactoferrin, transferrin (in body secretions, serum)
 bind free iron so inaccessible for bacteria
• HCl (in the stomach)
• Bile salts
 detergent effect disrupts bacterial cell walls
 6. Normal Flora
Skin:
• Associated with sweat glands:
• Mostly gram-positive bacteria
• Factors:
– Weather: warm & humid
– Age: young children
– Hygiene: unclean
 6. Normal Flora
Gastrointestinal Tract:

Ileum:
cell numbers of
105-107 per gram

Colon:
Cell numbers of
1010-1011 per gram
 6. Normal Flora
Urogenital Tract:
• Bladder: sterile
• Urethra: opportunistic pathogens
• Vagina: Lactobacillus acidophilus
ferments glycogen to produce lactic acid
& lower the pH of vagina
Stages in bacterial pathogenesis
1. Exposure to pathogen
2. Adherence
3. Invasion : usually via
mucosal surfaces / other
epithelium
4. Colonisation & growth:
proliferation
5. Toxicity : local or systemic
effect
Invasiveness: further
growth at original site or
distant site
6. Disease (Tissue damage)
PORTAL OF ENTRY
• Mucous membrane:
– Respiratory tract
– Gastrointestinal tract
– Genitourinary tract
• Skin
• Parenteral route
• Preferred portal of entry
 SPECIFIC ADHERENCE
• Bacterial adhesins (ligands):
– Capsule: dense polysaccharide or protein layer
closely surrounding a cell
– Glycocalyx: a loose network of polymer fibers
extending outward from the cell
– Slime layer: a diffuse mat of polymer fibers
surrounding cells that appear unattached to a
single cell
– Fimbriae
– Pili
• Receptors on host cells: sugars (manose)
COLONIZATION & GROWTH
• Factors:
– Capsules: impairing phagocytosis
– Components of cell wall, e.g.:
• M protein
• Wax
– Enzymes:
• Leukocidins
• Hemolysins
• Coagulases
• Kinases
• Hyaluronidase
• Collagenase
 INVASION
• Penetration into host cells:
– Invasins: rearrange nearby actin filaments of
cytoskeleton
Bacteria use actin to propel themselves through
the host cell cytoplasma (e.g. Shigella)
– Cadherin: glycoprotein which bridges
membrane junctions for bacteria to move from
cell to cell
– Fimbriae: adherence to host tissue
– Capsule: escape from phagocytosis
Salmonella Invasins
Shigella Invasins
 Fimbriae
• fimbria / pili
• Thread-like surface structures, 7 nm diameter,
0.5-2 mm long
• Repeating protein subunits, helical structure,
central pore
• 500 fimbriae per cell, 1000 protein subunits
• Promote specific adhesion via protein adhesins
• Overcome electrostatic repulsion
• Promote adhesion to target carbohydrate
receptors
Capsule
• Capsule not required for normal growth in vitro
• But required for survival in the host body – isolates
from invasive infection are encapsulated, but lose
capsule when subcultured in laboratory
conditions
• Major role in evasion of the host immune system -
protect from killing by complement and phagocytosis:
• weakly immunogenic, poor activators of
complement pathway
• inhibit opsonisation
• negative charge repels phagocytes
• shedding removes bound antibodies and
complement components
Bacterial toxicity
• exotoxins  secreted or membrane-bound proteins,
cause tissue damage or inhibit tissue
function

• endotoxin  lipopolysaccharide (LPS), component

of membranes elicits immune /
inflammatory responses
 Exotoxins
 Toxic bacterial proteins,
 Secreted
 Produced by a variety of bacteria (Gram + and -)
 Categories:
• Cytotoxin (neurotoxin, hepatotoxin, leukotoxin)
• Based on the species (tetanus toxin, cholera
toxin, diphtheria toxin)
• Based on the structure and functions (A-B toxins,
membrane disrupting toxin, superantigens)
 A-B Toxins mechanisms
 2 portions (Binding sub-unit and Active sub-unit)
 Connected by disulphide linkage
 B-subunit binds to specific host cell surface
molecules
 A-subunit is an enzyme, catalyses addition of
adenosine diphosphate-ribose (ADP-ribose) to the
target protein in the host cell, causing malfunction
of the cell
 e.g. cholera toxin, diphtheria toxin, tetanus toxin,
botulinum toxin, E. coli enterotoxin
 A-B Toxins e.g.

• Diphtheria toxin: disrupting protein synthesis by

blocking transfer of an amino acid from tRNA to
the growing polypeptide chain

• Botulinum toxin: preventing release of

acetylcholine from vesicles

• Tetanus toxin: preventing release of glycine from

vesicles
 Diphtheria toxin

Normal: Diphtheria toxin:

EF-2 (Elongation Factor 2)+tRNA Blocks EF-2
 Protein synthesis Protein synthesis stops
Cell death
 Botulinum toxin

Normal: Botulinum toxin:

Stimulation  acetylcholine release Blocks acetylcholine release
binds receptor no muscle contraction
induces muscle contraction Flaccid paralysis
 Tetanus toxin

Normal: Tetanus toxin:

Inhibitory interneuron  Glycine Blocks glycine release
 blocks excitation & no inhibition at acetylcholine
acetylcholine release  muscle release  irreversible
relaxation contraction  Spastic paralysis
 Cholera Toxin
• A-B toxin
• B sub unit  binds to GM1 ganglioside (a
glycolipid on the surface of epithelial cells)
• A sub unit   adenylate cyclase   cAMP
- inhibits sodium (Na) absorption
- activates secretory Chloride into lumen

Accumulation of Na + Cl intra lumen

osmotic imbalance  watery diarrhea
Normal
Cholera toxin mechanisms
Cholera toxin
 Membrane Disrupting Toxin
• proteins cause lysis of erythrocytes and other cell
types

• Lyses host cells by disrupting plasma membrane

- bind to cholesterol in target cell membranes,
insert as rings of oligomers forming pores (non-
enzymatic)
e.g. listeriolysin of Listeria monocytogenes

- act as phospholipase
e.g. -toxin / gas gangrene (C. perfringens)
- hyaluronidase
e.g. Group A streptococci
- elastase
e.g. Pseudomonas aeruginosa
Endotoxin
 Bacterial lipopolysaccharide (LPS)
• major part of the cell wall (Gram - )
• Unique, complex glycolipids that are an integral
component of the outer membrane
• Three structurally distinct elements
out in

n
O-antigen Core oligosaccharide Lipid A
Rough and smooth bacteria
• ‘Smooth’ bacteria - complete core and O
side chain

• ‘Rough’ bacteria - no O side chain, more

easily engulfed and destroyed by
phagocytes
 Endotoxin (LPS)
• Bacteria proliferate in blood – antibiotics effective if
given early

• Bacteria lysed by complement and phagocytosis,

LPS released into circulation

• Complement further activated, induces production

of cytokines

• Cytokines stimulate a massive inflammatory

response

• Results in septic shock- collapse of circulatory

system, multiple organ failure
Endotoxin (LPS)
LPS
Complement activation Macrophages

TNF

tissue damage Fever other inflammatory mediators

Toxic shock
References
Brock Biology of Micro organisms, 9th edition,
2000, Madigan et al. page 773 - 800

Medical Microbiology and Immunology, 7th edition,

2002, Warren Levinson and Ernest Jawetstz, page
27 - 44
 5 4
Gs
GTP =
guanosine
Gs
1 triphosphate
Gs
2 3
Gs

Normal:
Gs protein (with normal GTPase) + GTP
Cannot
+ adenylate cyclase Gs-GDP stimulate
adenylate
Na+ movement
 cAMP cyclase
from lumen to
blood stimulatory process
stopped
 Mode of action of
cholera toxin Gs
Gs 
2
1 3
Gs


Cholera toxin:
Cholera toxin-Gs protein + GTP

+ adenylate cyclase No conversion to GDP

 cAMP Ion and fluid flux

across intestinal Watery diarrhea
membrane disrupted