Lecture 5: Parasite glycobiology Flashcards

1
Q

True or false: trypanosomatids all have a kinetoplast?

A

True

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2
Q

Which trypanosoma species is Leishmania clade closely related to?

A

Trypanosoma cruzi and Trypanosoma brucei

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3
Q

What disease is caused by T. brucei?

A

Human African Trypanosomiasis (HAT) or Sleeping sickness

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4
Q

What is the vector for the following parasites:
1) T. brucei
2) T. cruzi
3) Leishmania sp.

A

1) Tsetse fly

2) Triatomine bug

3) Sandfly

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5
Q

Which form of the parasite is taken up by the Tsetse fly and which form of the parasite is injected by the Tsetse fly into the human?

A

Taken up: stumpy form that rapidly differentiates to procyclic form
Injects: metacyclic form

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6
Q

True or false: T. brucei is strictly extracellular?

A

True

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7
Q

True or false: Leishmania sp. is strictly extracellular?

A

False, they are intracellular (phagocytosed by macrophages and proliferate within parasitophorous vacuole as amastigotes)

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8
Q

How does the triatomine bug transmit the T.cruzi parasite to the human host?

A

Bites around eyes and mouth (soft tissue parts), defecates parasite close to wound site, which is irritated and the parasite is rubbed into the wound

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9
Q

True or false: T. cruzi is strictly extracellular?

A

False: it is intracellular - cross epithelial barrier so can enter tissues (mainly smooth muscles particularly of the heart, skeleton and oesophagus)

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10
Q

What is the name of the disease caused by T. cruzi?

A

Chagas disease

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11
Q

what does the surface coat of the T. brucei parasite contain in the human host?

A

mainly VSGs

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12
Q

what does the surface coat of the Leishmania parasite contain in the human host?

A

LPG
Surface proteases (PSP)
GIPLs

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13
Q

what does the surface coat of the T. cruzi parasite contain in the human host?

A

Mucins
Trans- sialidases
GIPLs

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14
Q

What are the T. brucei glycoconjugates and what is the structure of each?

A

VSG = homodimer, each monomer is N-glycoslyated and GPI anchored to the plasma membrane

Transferrin receptor = heterodimer, both monomers are N-glycosylated but only one monomer is GPI anchored to the plasma membrane

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15
Q

True or false: the T. brucei transferrin receptor (TfR) is structurally related to VSG?

A

True, expressed form the same expression site (ES) by RNA pol1

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16
Q

What is the role of the T. brucei TfR?

A

heterodimer that recognises and internalises host transferrin

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17
Q

What is the role of VSG and TfR glycosylation in T. brucei?

A

shield the plasma membrane from complement and prevent exposure of conserved GPI anchor

18
Q

What glycoproteins are expressed in the surface of the T.brucei parasite in the Tsetse fly mid-gut?

A

Procyclins (Aka procyclic acidic repetitive proteins (PARPs)

19
Q

Describe the structure of procyclins in T brucei

A

GPI-anchor that is heavily modified by addition of long glycan chain that masks the surface
Contains a single N-glycosylation site for GlcNAc2Man5 glycan
- pack together to form a dense surface coat.
- multiple procyclin isoforms that are classified as either EP (containing EP repeats) or GPEET (containing GPEEpT repeats) procyclins.

20
Q

How does the expression of procyclin isoforms differ during tsetse transmission?

A

Developmentally expressed
- GPEET expressed in early infection
- EP expressed in late infection

21
Q

How do procyclins promote survival of parasite in tsetse fly midgut?

A

N-terminal domain is cleaved by fly proteases and the remaining glycoconjugate is protease-resistant and provides a tough coat against gut hydrolases

22
Q

How does procyclin expression allow the T. brucei parasite to be injected into the host?

A

Once in midgut, the parasite needs to migrate across the peritrophic matrix into the salivary glands to be injected back out
- procyclin-null mutants are not as able to escape from the mid-hut into the salivary glands

23
Q

Describe the structure and function of the Leishmania LPG?

A

GPI-anchored phosphoglycan that dominates the cell surface of the Leishmania parasite

  • GPI-anchor (conserved)
  • glycan core (conserved)
  • repeat unit made of mannose and phosphorylated galactose which may or may not be bound by different sugars (hexoses or pentoses) - variation
  • cap (variation)

Function:
- required during sandfly transmission
- prevent lysosome-phagosome fusion during metacyclic-to-amastigote differentiation

24
Q

Describe the structure and function of the Leishmania PSP?

A

GPI-anchored glycoprotein and is a protease widely known as GP63

important for the phagocytosis of metacyclic Leishmania by host macrophages - part of invasion process

25
Q

What are GIPLs on the Leishmania parasite surface and what is their role?

A

Small glycoinositolphospholipids

have role in protection for the otherwise exposed plasma membrane

26
Q

Where does variation in the Leishmania LPG phosphoglycan arise?

A

variation in glycosylation pattern and length of the repeat unit between species, lifecycle stages and strains

27
Q

What is the function of Leishmania LPG in the promastigote stage?

A

required for attachment to mid-gut epithelium in the sandfly, which is mediated by the insect lectin (galectin) as the receptor for the LPG ligand (recognises surface galactose)
- if attachment does not occur, parasite would be excreted

28
Q

How does the Leishmania LPG structure change as they differentiate form the procyclic to the metacyclic promastigote form in the sandfly midgut?

A
  • cap the galactose residues with different sugars so they are no longer able to attach to the galectin in the sandfly midgut
    –> this allows them to be injected with the blood meal into the human host
29
Q

The difference in Leishmania LPG strucute is thought to define what in terms of sand fly species and the human host?

A

Thought to define:
- the interaction of Leishmania species with different sandflies
- the interaction and severity of disease within the human host

30
Q

What is Leishmania PSG?

A

Promastigote secretory gel - a mix of extracellular glycoconjugates with a mucin-like filamentous proteophosphoglycan (fPPG) as a major component
- secreted in the anterior mid-gut to form a gel-like substance within which infectious metacyclic promastigotes accumulate

31
Q

Which form is the Leishmania parasite in when it is uptaken by the sandfly?

A

Amastigote (then differentiates to procyclic promastigote that attached to mid-gut, then differentiates to metacyclic promastigote that allows it injection again into the mammalian host)

32
Q

What is the major component of Leishmania PSG?

A

filamentous proteophosphoglycan (fPPG)

33
Q

What is the function and role of Leishmania PSG?

A

secreted in anterior mid-gut by metacyclic promastigotes which forms a plug that blocks flies from feeding effectively
–> alters the feeding behaviour of the sandfly - results in continued re-probing of the host as it cannot take a full blood meal so thinks it hasn’t been able to inject into somewhere where it can feed
- eventually regurgitates the PSG plug containing metacyclic promastigotes in order to increase feeding
–> result in increased potential for parasite transmission

34
Q

What role does Leishmania PSG play in infectivity within the mammalian host?

A

role in the infectivity within the mammalian host
- inoculation of mice with parasite with addition of PSG results in increased size of lesion and infectivity
- glycan moieties on glycosylated fPPG is sufficient for lesion exacerbation

35
Q

What component of the fPPG is responsible for increased lesion size and infectivity in mammalian host?

A

the glycan moieties on glycosylated fPPG (but not eh peptide backbone of fPPG)

36
Q

What is the major component of the T. cruzi surface coat and what is their structure?

A

Mucins
- glycoproteins rich in Ser/Thr with multiple O-linked glycans

37
Q

Explain the expression of mucins in T. cruzi

A

Mucins are encoded by an expanded multi-gene family formed of 800 genes
- the different classes of mucins are developmentally regulated (different classes expressed in the epimastigotes and mammalian forms)

38
Q

True or false: Trans-sialidases in T. cruzi are encoded by an expanded multi-gene family?

A

True

39
Q

What are the three main processes in which T. cruzi mucins are involved in?

A
  1. Protection
  2. Attachment
  3. Host cell invasion
40
Q

How are T. cruzi mucins modified?

A

Addition of host sialic acid by the parasite derived trans-sialidase bound to cell surface
- trans-sialidase transfers sialic acid from host serum sialoglycoconjugates to the mucins on their surface

41
Q

What is the purpose of sialic acid modification of T. cruzi mucins?

A

sialic acid is quite specific to mammalian cells so addition of sialic acid to mucins allows evasion of immune system and recognition by host immune system as ‘self’

42
Q

What are the three different trans-sialidases of T.cruzi and how do they differ?

A

aTS = active trans-sialidases
iTS = inactive trans-sialidases
soluble aTS

aTS transfers sialic acid from host serum sialoglycoconjugates

iTS recognises and binds sialic acid but does not transfer it - this is part of the recognition of host cells for invasion process to occur

soluble aTS is used to cause confusion - transfer sialic acid from host serum glycoproteins and remodel the host glycophenotype to cause immune response to self cells - can not distinguish between normal glycophenotype and the parasite. - escape detection from immune system whilst it becomes intracellular