Host-pathogen interaction

bacterial side of the story

Saji George
 Facts on host-bacteria interaction
Both the host and bacteria have evolved
numerous ways in which to
communicate their wishes/intensions
with each other.

The communication between host and

bacteria is essential for the survival of
both

Often bacterial side of the story is

neglected
Pathogen Disease Host cell
Interaction
Extracellular:
- S.aureus Skin/Tissue Adherence to ECM
- S.pneumoniae Pneumonia Adherence to ECM
- B. pertussis Whooping cough Adherence to cells
- N. gonorrhoeae Gonorrhea Adherence to cells
- E.coli UTI, diarrheas, meningitis Adherence to cells
- H.pylori Ulcers, gastritis Adherence to cells

Intracellular
Macrophages:
- L. pneumoniae Legionaires’ disease Within vacuole
- M. tuberculosis Tuberculosis Within vacuole
Macrophage &
Epithelial cells
Typhoid fever, gastroenteritis Within vacuole
- Salmonella species
Dysentery, gastroenteritis Intracytoplasmic
- Shigella species
Listeriosis, meningitis Intracytoplasmic
- L.monocytogenes
Trachoma, STD, pneumonia Within vacuole
- Chlamydia species
 Stages in host-bacteria interaction-disease
condition

1. Adherence
2. Invasion
3. Initial multiplication
4. Evasion of defenses
5. Spread of infection
6. Damaging the host
 Prokaryotic and Eukaryotic Interactions

Eukaryotic Cell Pili or adhesins Prokaryotic Cell

Virulent Bacteria

Receptor
 Prokaryotic and Eukaryotic Interactions
Eukaryotic Cell Pili or adhesins Prokaryotic Cell

Virulent Bacteria

Receptor

COLONIZATION
 Prokaryotic and Eukaryotic Interactions
Eukaryotic Cell Pili or adhesins Prokaryotic Cell

Virulent Bacteria

Receptor

COLONIZATION INVASION
 Adherence to host cell- First step in the
host-pathogen interaction

Site of bacterial adherence

Keratinized or mucosal epithelium

Specialized epithelium (ocular, aural)

Cell membrane components

 Why do bacteria adhere?

To avoid physical and Adherence often combined

immunological removal, with manipulation of host
bacteria must adhere to cell signalling and
– cell surfaces and extracellular
matrix
cytoskeleton
e.g. in respiratory, – Invasion
gastrointestinal and – Intimate adherence
genitourinary tracts
– solid surfaces
e.g. teeth, heart valves,
prosthetic material
– other bacteria
 Molecular aspects of Host-
Pathogen interaction

Mechanisms of Adherence to Cell or

Tissue Surfaces

1.  nonspecific adherence: reversible

attachment of the bacterium to the
eukaryotic surface

2. specific adherence: irreversible

permanent attachment of the
microorganism to the surface
Nonspecific adherence involves nonspecific attractive forces
which allow approach of the bacterium to the eucaryotic cell
surface.

1. Hydrophobic interactions
2. Electrostatic attractions
3. Atomic and molecular vibrations resulting
from fluctuating dipoles of similar
frequencies
4. Brownian movement

Specific adherence.

1. Irreversible attachment of
bacteria to host
2. Mediated by bacterial surface
appendages such as pilus fimbrae
etc
Molecular mechanism of host bacteria interaction

PAMPs

Pathogen-associated molecular
pattern

A limited set of conserved

molecular patterns that are
unique to microbial world and
invariant among entire classes
of pathogens
 Adherence mechanisms

General adherence mechanisms

– Capsules and slime
– Biofilm formation

Gram-positive adhesins (PAMPs)

– MSCRAMMs (microbial surface components
recognizing adhesive matrix molecules), e.g.
protein A
– Fimbriae

Gram-negative adhesins (CHO and protein

receptors) (PAMPs)
– Fimbriae, Afimbrial adhesins (FHA, Pertactin
etc.)
– Outer Membrane Proteins
– Types III-IV secretion
Molecular mechanism of host bacteria interaction

Pattern Recognition Receptors

Carbohydrate residues of glycoproteins or
glycolipids (Protein-carbohydrate interaction)
- This binding is quite specific

Extracellular matrix proteins. (Protein-protein

interaction)
- I.e. Fibronectin

In some cases the pathogen injects its own

protein receptor into the host cell.

It is common for a pathogen to express and

utilize more than one adhesin
Host Cell Receptors for Adhesins-PRR
 Molecular mechanism of host bacteria interaction

Toll-Like Receptor 4: first human PRR to be identified

Toll: protein from D. melanogaster identified for its role in dorso-ventral

embryo patterning and critical for effective immune responses in adult
flies against the fungus Aspergillus fumigatus
 C-reactive protein is a PRR
CRP does not act alone
but collaborates with
other plasma PRRs to
form stable pathogen
recognition complexes
when targeting a wide
range of bacteria for
destruction.

Ng et al. The EMBO Journal (2007), 1–10

TLR detect multiple PAMPs

Annu. Rev. Cell. Dev. Biol. (2006) 22: 409-37

 Adhesion factors of pathogenic bacteria

adhesion factor examples

Type-I-fimbriae enterobacteria
Type-IV-fimbriae N. gonorrhoeae
Fimbrial adhesins V. cholerae
(multi-subunit adhesion P. aeruginosa
organelle) P-fimbriae uropathogenic E. coli
S-fimbriae uropathogenic E. coli
M-protein S. pyogenes
curli E. coli
Neisseria meningitidis in meningococcal
pathogenesis
(i) initial adhesion
Is mediated by pilus

(ii) intimate adhesion

Other bacterial and cellular
Structure. Pilus disappear during
the intimate adhesion

Transcriptional regulation at the

initial adhesion is required for
the intimate adhesion. crgA
gene is up-regulated on initial
contact

Deghmane et al The EMBO Journal Vol.19 No.5 pp.1068–1078, 2000

 Adhesion factors of pathogenic bacteria

Non-fimbrial
adhesins NFA, AFA E. coli
(multi-subunit)
LPS N. gonorrhoeae
Polysaccharides LOS N. meningitidis
EPS P. aeruginosa
PIA (= polysaccharide adhesin; ica) S. epidermidis
Biofilm formation
alginate P. aeruginosa
 Invasion of the host cell

Once adhered to a host surface, some pathogens gain deeper

access into the host (invasion)

This process can be divided into two types:

Extracellular invasion – bacteria breaks down the
barriers of a tissue to disseminate in the host –
Secretion of several enzymes that degrade host cell
molecules
Intracellular invasion – penetrate the cells of a host
tissue and survive within this environment.
Invasion- extracellular and intracelular
-Urinary tract infection
 Invasion-Adhesins
S. aureus in Invasin
endothelial cells Integrin receptors
a
Yersini 






actin


       

extracellular matrix


actin
   

S. aureus invasion depends on:






1. fibronectin Yersinia





2. Fibronectin binding protein of
  
  

S. aureus 
3. 51-integrins as receptor on 


the host cell

4. actin polymerization
extracellular matrix
 Mechanisms used by bacteria to enter cells.

(A) The zipper

mechanism
used by
Yersinia and
Listeria.

(B) The trigger

mechanism
used by
Salmonella
and Shigella
 Zipper mechanism
(i) Contact and adherence. Invasin
This step is independent of the actin
cytoskeleton and involves only the Integrin receptor
a
bacterial ligand and its receptor. It Yersini 

leads to receptor clustering.






actin


(ii) Phagocytic cup formation.
This step is triggered by the transient        
signals occurring after formation
of the first ligand-receptor extracellular matrix
complexes and propagating
around the invading microbe. 

These signals induce actin

polymerization and membrane
actin

extension.   





Yersinia


(iii) Phagocytic cup closure and



retraction, and actin
  
  

depolymerization.




extracellular matrix
 Trigger mechanism
Invasion of salmonella
Salmonella enters host cells by
inducing host cell membrane
ruffling
membrane ruffles non-specifically
wrap around the bacteria and pull
them into the cell
Salmonella end up in membrane-
bound vesicles called Salmonella-
containing vacuoles (SCV). 
SCVs are unique environments
within the cell defined by the
bacteria within them
As they mature, SCVs do not
follow the defined routes of
cellular trafficking of vesicles and
differ in their composition from
normal phagosomes
1. A pre-interaction stage.
Effector molecules are stored in
bacterial cytoplasm
Type three secretion systems are
properly formed

2. An interaction stage

3. The formation of a
macropinocytic pocket.

4. Actin depolymerization and

closing of the
macropinocytic pocket
 Secretion of bacterial products- the
secretion systems
Type I, II, III, IV, V and VI

Type I, III and IV secretions is directly

from bacteria to eukaryotic cells

Type II and V- two-step processes in

which proteins are transported first
through the inner membrane (IM) and
then through the outer membrane
(OM).
Type III secretion system is necessary
for invasion.
Type VI mechanism of secretion
Naturally produced OM vesicles from
pathogenic bacteria contain adhesins,
toxins, and immunomodulatory
compounds.
Mediate bacterial binding and
invasion, cause cytotoxicity, and
modulate the host immune response.
 Type III secretion system

Type III Secretion Systems

are multi-protein complexes
connecting bacteria to host
cells
Mediate protein secretion
and translocation from
bacterial cytoplasm to host
cell interior
Effector proteins subvert
cellular functions
Modulation of Virulence Factor Expression by
Pathogen Target Cell Contact
Yop is a virulence factor produced by Y.
pseudotuberculosis
Yop (Yersinia outermembrane protein, is an inhibitor of
NF-kß signaling)
Contact with HeLa cells provoke Y. pseudotuberculosis
to produce yop proteins into the cytoplasm
Yop is secreted by type III mechanism
Shigella shows target cell induced production of Ipa
proteins
Salmonella forms surface structures on contact with
target cells
The type III secretion is blocked when bacteria is present with Ca2+
Contact between the target and bacteria opens up the secretion
channel at the zone of contact
This promotes the bacteria to produce yop proteins.
 Salmonella-invasion of epithelial cells

Appendages are required for bacterial

entry into epithelial cells

Only salmonella in contact with epithelial

cells produced surface appendages Bacteria that are inside the
membrane ruffles lack the
surface appendages (second
step)
 E. faecalis entry into HeLa cells

Two mechanisms are involved in the entry of

E. faecalis into HeLa cells

One, sensitive to amiloride, is most likely a

macropinocytic, actin-dependent uptake
mechanism, resulting in the production of large
smooth membrane vacuoles engulfing
enterococci.

The other is receptor mediated entry (RME), in

which entry is dependent on both MF and MT
structural integrity.

Bertuccini et al. Med Microbiol Immunol (2002) 191: 25–31

Ligand-Signaled Upregulation of Enterococcus faecalis
ace Transcription, a Mechanism for Modulating Host-E.
faecalis Interaction
Sreedhar R. Nallapareddy and Barbara E. Murray

mRNA of Ace
protein increased in a
dose dependent
manner as the
concentration of
collagen was added
to growth medium
(B) Dose-dependent induction of ace
expression by OG1RF.
The adherence of E. faecalis (OG1RF) to plastic plates
coated with collagen increased once bacteria were grown
in presence of collagen and serum.
 Summary
Bacteria interact with eukaryotic cells via
molecular mechanisms

The interaction of bacteria with eukaryotic

cells could trigger the expression of
virulence factors that ultimately
determines its persistence or perish
Thank you
 Virulence Factors of S. mutans

Specific adherence to tooth surface using

antigen I/II adhesin and GTF

Production of extracellular
polysaccharides (dextran)

Accumulation of intracellular amylopectin-

like polysaccharides (carbon/energy
reserve)
Tooth Decay Dental Carries
Bacteria’s Lactic acid
assembles break down
Perfect Cannot on the enamel in
caused attach to
example dextran to the areas of
by a clean
is form plaque the plaque
Strep mutans teeth

Can attach
to a pellicle Glucosy- Fructose
within transferase undergo
Pellicle is assembles metabolism
minutes
protein glucose into resulting in
form dextran lactic acid
from
saliva
Hours later
cariogenic
bacteria
E.g.... S mutans
can occupy
will hydrolyze
this pellicle
sucrose into
G+F