skin barrier models-molluscum
TRANSCRIPT
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MCV biopsy
NIH Clinical Center: Bugert and Turner, 1993
www.dartmouth.edu/~thabif/ weeklyclinic111901/..
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MCV histopathology
Lobular hyperplasia of epidermis into the dermis resulting in a cup-shaped lesion. The basal layers appear normal, but the keratinocytes of stratum malpighii become enlarged and acquire eosinophilic inclusion bodies - Henderson-Patterson bodies that contain virus particles.
From biopsy; University of Heidelberg: Bugert, 1988
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MCV in the host cell
SEM Shelley and Burmeister, 1986
MCV can be purifiedin large quantities from skin biopsies
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Abb. 1.1, Schemazeichnung eines Pockenpartikels
Tubuli
NukleosomLateralkörper
Aussenmembran
Hülle
Core Membran
100nm
MCV virus preparation ELMI: 220- 450 nm filter
MCV-HD-18-yellow-Cardiff 5
350 nm
MCV-HD-10-red-Cardiff 1
Attempt to grow
in cells in culture
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100nm
1. only human host / no cell culture system
MCV infection - biological features
2. epidermal tissue tropism (gene acquisition-adaptation)
3. epidermal dedifferentiation
4. epidermal hyperproliferation-benign tumors
5. immuneevasion: lack of cellular immune response
6. cell receptor overexpression (EGFR and transferrinR)
7. HPV, EBV (analogy to other human epithelial viruses)
α EGF receptor
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MCV replication in the epidermis
n
no egress ?
Stratum corneum
Keratinocyte
dna
e-rna
virus ?
epid
erm
is
UV
vegetative replication ?
cytoplasm
?
MCV
mc148R
mc159/160R
mc054L
mc066L
MCV growth characteristics in situ
3. Keratinocyte dedifferentiation↓ keratin
6. Speed-up of cell migration25 days instead of 33
9. Enhanced mitosis rateincreased cell proliferation
→tumor formation
some proteins involved...........
• mc159/160-FLICE inhibitors (↓apoptosis)
• mc066-glutathione peroxidase (↓UV-induced apoptosis)
• mc007-RB binding protein (cell cycle: G-S phase)
MC
V t
umor
igen
esis
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Minucell®-Loop 3
Minucell ® dual perfusion chamber cell culture system
60.45µM
0.45µM
0.45µM
sterile filter
medium reservoir
peris
talti
c pu
mp
37 °C heater plate
sterile filter
Gas expander module
sterile filter perfusion chamber
sterile filter
3 way valve (injection port)
microscopy
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Emma Smith: Minucell system in operation (Rm. 115) 9/2004
injection / sample port