Opportunistic infection of the central nervous system by human polyomavirus
TC can cause a devastating disease, progressive multifocal leukoencephalop
athy (PML). To gain new neuropathological insights into TC-virus (JCV) infe
ction patterns in PML at the light microscopic level, the highly sensitive
indirect in situ polymerase chain reaction (in situ PCR) was employed in up
to 15-year old formalin-fixed and paraffin-embedded postmortem brain tissu
e derived from nine AIDS patients with PML. In situ PCR, in which target DN
A is amplified intracellularly and detected by a specific labelled probe in
morphologically intact tissue, was compared with conventional in situ hybr
idization (ISH). Validity was ensured by the inclusion of 13 controls. JCV
detection with in situ PCR proved to be highly sensitive since in all nine
brain samples the number of positive cells exceeded the ISH results by 2-3-
fold. Whereas by routine staining the brain tissue of each individual patie
nt showed regions with severe, mild or no involvement by PML, improved dete
ction of JCV DNA by in situ PCR allowed a regrading into five different deg
rees of JCV infection. Significant myelin staining was observed, suggesting
that cell-to-cell contact may not be the only means of virus spread but th
at new cells could also be infected by virus released after cell lysis. Fur
thermore, using in situ PCR hitherto unreported intracellular distribution
patterns of JCV DNA in oligodendro- and astrocytes were observed by light m
icroscopy.