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Infection and Immunity > 24 In vivo imaging of fungal infections > Flashcards

24 In vivo imaging of fungal infections Flashcards

1
Q

Definition of in vivo Imaging

A

non-invasive technique for investigation of cells/pathogens in living animals

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2
Q

how is virulence investigated

A

in vivo imaging

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3
Q

what are the advantages of in vivo imaging

A
  • Repeated investigation of individual animals
  • Monitoring of pathogen and/or immune cells in temporal and spatial resolution
  • Insights on disease progression from small animal groups
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4
Q

what are the disadvantages of in vivo imaging

A

Clinical and environmental isolates require genetic modification (reporter strains)
„High-throughput” not possible due to time-consuming measurements

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5
Q

what are the common techniques in in vivo imaging

A

Light-based
Fluorescence-Imaging
Bioluminescence-Imaging

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6
Q

what are the advantages of fluorescence imaging

A
  • Large selection of fluorescent reporters available

- Different excitation and emission wavelength allows multiple labellin

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7
Q

what are the disadvantages of fluorescence imaging

A
  • High background fluorescence from animals à low sensitivity
  • Low-wavelength excitation reporters difficult to activate within tissues
  • Small band-width for detection of emission spectra (overlap with excitation)
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8
Q

Bioluminescence imaging

Advantages

A
  • Hardly background luminescence from animals

- Very high sensitivity

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9
Q

Bioluminescence imaging disadvantages

A
  • Small number of reporters available
  • Frequently substrate injection required
  • Biochemical reaction required for light production that might affect fitness
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10
Q

what are the Luciferase reporters

A

Firefly, Renilla and Gaussia commonly used in eukaryotes

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11
Q

what is the lux operon not suitable in

A

bacterial “lux” operon

Not suitable for eukaryotic systems

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12
Q

what does Gaussia luciferase possess

A

very high quantum yield

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13
Q

what is the adavantage of secretion - Gaussia luciferases

A

Secreted luciferase can be detected from fluids

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14
Q

what is the disadvantages of secretion - Gaussia luciferases

A
  • Secretion is not quantitative

- Difficult to detect infection foci

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15
Q

Firefly luciferase for in vivo imaging advantages

A

Substrate D-luciferin highly water soluble = Transported to all body sites
Light emission in vivo = Reduced absorption by haemoglobin
Codon-optimisation of reporter for optimal sensitivity

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16
Q

what is an example of codon optimisation of luciferase sequences

A

Example on the filamentous fungus Aspergillus fumigatus

17
Q

what is Candida albicans like

A

dimorphic (polymorphic fungus)

18
Q

where does dissemination occur in candida albicans

A

host via yeast cells

19
Q

how does tissue invasion occur in candida albicans

A

by hyphal filaments

20
Q

what infection does candida albicans cause

A

Superficial infections
Invasive Candida albicans infections
Nosocomial acquisition

21
Q

who gets Candida albicans

Superficial infections

A

3/4 of all healthy women experience at least one vaginal yeast infection
Oral Candida infections affect > 90% of HIV infected patients

22
Q

how frequent are Candida albicans infections

A

Candida sp. are the third most frequent nosocomial bloodstream isolates in the USA

23
Q

what does Firefly luciferase work as

A

reporter gene

24
Q

how does in vivo investigation of C. albicans pathogenicity occur

A

Generation of bioluminescent C. albicans reporter strains

Selected promoter

25
Q

how is extra-renal sites of infection detected

A

Ex vivo
Analysis of mice with signals outside the kidney localisation
Mouse 4 displays an additional signal

26
Q

how is the suitability of reporter strains for disease monitoring

A

= Disease progression monitored from individual mice
= Major and minor sites of infection detected
= Light intensity correlates with fungal burden

27
Q

what is the meaning of luminescence in evalution

A

Signals decrease to background under therapy

28
Q

How can C. albicans persist in the gall bladder under therapy

A

Bile confers resistance against Caspofungin

29
Q

can other antifungal drugs persist in gall bladder under therapy

A

Bile confers a decrease in susceptibility against a wide range of different antifungal

30
Q

what is BLI

A

powerful tool to study fungal infections

31
Q

what does the selection of luciferase depend on

A

study aims

32
Q

what are firefly luciferase best suited for

A

deep-seated infections

33
Q

what does bile form

A

micelles

34
Q

what is the effect of micelles

A

reduced bioavailability of drugs

35
Q

what may gall bladder form

A

cryptic reservoir for re-infection

Infection and Immunity (29 decks)

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