Enteropathogenic and Enterohaemorrhagic E.coli Flashcards

1
Q

What age group is EPEC most common and what is the mortality rate?

A

<2 years with a >30% mortality rate

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2
Q

What are symptoms, Route of infection and reservoirs of infection for EPEC?

A
  • Diarrhoea, Vomiting, Fever
  • Route of infection: Faecal-oral transmission, contaminated surfaces, infant food, water, hands
  • Reservoirs of infection: Asymptomatic and symptomatic children, asymptomatic adult carriers, mothers and persons handling children
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3
Q

Does EPEC produce toxins? Is it invasive?

A

No, Non invasive.

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4
Q

How is EPEC defined? (by the presence of…)

A

Presence of LEE and Bfp

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5
Q

What is the difference between a typical EPEC and an atypical EPEC?

A

Typical EPEC has bfp atypical does not

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6
Q

How does EPEC attach and efface lesions? (3 stages)

A
  • Stage 1: Non-intimate association via bundle forming pili
    ○ Bacterial cells aggregate together
  • Stage 2: Type 3 secretion of Tir and other effectors
    ○ Assembly of Type III secretion system which
    leads to intimate adherence
    Stage 3: Intimate adherence, Attaching and Effacing lesion
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7
Q

What is a Bundle forming pili? How is It regulated? What is it’s importance in virulence (EPEC)

A
  • Mediates bacterial-bacterial interaction -> forming of microcolonies
  • Transcription of bfp regulated by Per A
  • Encoded by PerABC operon on EAF plasmid
  • Per B and C required for full transcriptional activation of the bfp gene cluster
  • Importance in virulence: mediated initial adherence to host through n-acetyl-lactosamine receptors on host cell
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8
Q

What is a pathogenicity island?

A
  • A segment of a plasmid with genes encoding for virulence factors
  • Usually large but unstable due to ‘mobility’ genes
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9
Q

What does LEE stand for and do? How many operons are there? Why is it necessary? (EPEC)

A
  • Locus of Enterocyte Effacement (LEE)
  • 5 operons, in order - 1,2,3,5,4
  • Necessary for the attaching and effacting effect and pedestal formation
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10
Q

What does each Lee operon encode? (EPEC)

A
  • LEE1-3
    ○ Encodes type 3 secretion system (TTSS) for
    translocating bacterial proteins towards the
    enterocyte
  • LEE4
    ○ Encodes esp genes (EPEC secretion proteins)
    espADB encode for translocator proteins that form
    a conduit through which T3SS delivers effector
    proteins to host cell
  • LEE5
    ○ Encodes genes necessary for bacterium-host cell
    intimate adhesion (eae: encodes adhesin intimin,
    tir: Translocated intimin receptor)
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11
Q

How do EPEC secrete effector proteins into host cells using the T3S system?

A
  • Inner and outer ring components assembled
  • EscF polymerized and assembled into outer and inner rings
  • EspA filament secreted via type 3 secretion system and attached to the tip of the needle
  • Formation of a hollow filament
  • Translocation effector proteins enter into host cell
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12
Q

What is the role of T3SS in intimate adherence? (EPEC)

A
  • Intimin used to cause intimate binding, however there is no natural receptor for it
  • Tir (translocated intimin receptor) is translocated into host cell via T3SS and acts as a receptor for intimin
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13
Q

How does Tir activate an actin signaling cascade? (EPEC)

A
  • Tir becomes tyrosine phosphorylated in host cell plasma membrane
  • Binds to adaptor protein
    ○ Nck
  • Nck recruits N-WASP or a WIP-N-WASP complex
    ○ Triggers activation of the Arp2/3 complex
  • Actin is assembled in a pedestal formation
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14
Q

How does EPEC cause diarrhea?

A
  • Damage to absorptive apical surface causes failure to absorb water
  • Injected effector proteins disrupt tight junctions
    ○ Increases permeability of mucosal enterocyte tight
    junctions
    ○ Activation of NFkb
    ○ Activation of IL-8 cytokine
  • Recruitment of polymorphonuclear neutrophils (PMNs)
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15
Q

What diseases can EHEC cause?

A
  • Diarrhoea

- Bloody stools

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16
Q

What is the origins of an EHEC?

A
  • Originally an atypical EPEC which acquired a shiga-like toxin via a bacteriophage
17
Q

Explain the pathogenesis of EHEC

A
  • Similar to EPEC
  • LEE pathogenicity island present
  • Forms A&E lesions
  • Produces large amounts of shiga-like toxin which acts on sites remote from intestine
18
Q

Explain EHEC and adhesion?

A
  • No bfp
    ○ Uses wide variety of adhesions
  • Mediate similar binding and actin rearrangements as EPEC
  • Use Intimin (eae) adhesion to intimately attach to host cells
19
Q

Explain actin signalling cascade activated by EHEC during pedestal formation

A
  • Tir localizes to the plasma membrane
    ○ NOT tyrosine phosphorylated
  • Tir binds IRTKS/IRSp53
  • The EHEC effector EspFu links complex to N-WASP
  • N-WASP stimulates Arp2/3 nucleation of actin
    Pedestal formation
20
Q

What toxin does EHEC produce? What are the 2 immunologically distinct types and how do they differ?

A
  • Shiga-like toxins
  • Stx1, st2
    ○ Stx2 is associated with severe human disease
21
Q

How are Shiga-like toxins encoded? (EHEC)

A

Encoded within lysogenic bacteriophage

22
Q

Explain the structure of Shiga-like toxins (EHEC)

A

Ab5 type toxin, active part central with b part surrounding

23
Q

What is the receptors for Shiga-like toxins (EHEC)

A

Gb3

24
Q

Explain the mechanism of action of Stx toxin (EHEC)

A
  • In kidney B subunit of toxin binds to Gb3 which is expressed on cell surface
  • Shiga-like toxin damages blood vessels and kidney cells
  • Action of damage is:
    ○ External toxin enters clathrin coated pit
    ○ Coated vesicle is formed
    ○ Fusion of lysosome with coated vesicle
    ○ Toxin is processed in cytosol
    ○ A1 cleaves 70S ribosomes which inhibits protein
    synthesis
25
Q

What is Haemolytic Uremic Syndrome? (EHEC)

A
  • Stx can also activate platelets
  • HUS is when there is a deposition of microthrombi on damaged kindey capillary endothelial cells which leads to occlusion and reduced O2 -> renal failure