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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