Three cases of human herpesvirus-6 latent infection: integration of viral genome in peripheral blood mononuclear cell DNA.
Saliva and peripheral blood mononuclear cells from three patients, two with lymphoproliferative disorders and one suffering from multiple sclerosis, were examined for the presence of human herpesvirus-6 (HHV-6) genome by using the polymerase chain reaction and Southern blot analysis. The search for anti-HHV-6 antibodies, carried out in the sera of the same cases by an immunofluorescence assay, was negative in two cases at the lowest dilution used (1:40). These three patients had a high number of HHV-6 specific sequences in uncultured peripheral blood mononuclear cells, which are thought to be a normal site of viral latency although, in healthy individuals, the infected cells are extremely rare. In order to gain some insight into the state of the viral genome in this latent HHV-6 infection, we used pulsed field gel electrophoresis to separate HHV-6 DNA directly from HHV-6 (strain GS) infected HSB-2 cells and from the peripheral blood mononuclear cells of these three patients. Our study showed the presence of intact viral genome, of the expected length of 170 kb, persisting as free extrachromosomal element in the HSB-2 cells but not in patients' peripheral blood mononuclear cells. On the other hand, in strong contrast with the results obtained in infected HSB-2 DNA, the restriction analysis of the three patients' peripheral blood mononuclear cell DNA showed fragments of molecular weight constantly higher than the 170 kb segment, indicating that the viral sequences are linked to high molecular weight cellular DNA.(ABSTRACT TRUNCATED AT 250 WORDS)
Luppi
M
M
Center for Experimental Hematology, University of Modena, Italy.
Marasca
R
R
Barozzi
P
P
Ferrari
S
S
Ceccherini-Nelli
L
L
Batoni
G
G
Merelli
E
E
Torelli
G
G
D000328:Adult; D000914:Antibodies, Viral / Q000097:blood; D015139:Blotting, Southern; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D016521:Electrophoresis, Gel, Pulsed-Field; D005260:Female; D005455:Fluorescent Antibody Technique; D015321:Gene Rearrangement; D016679:Genome, Viral; D006566:Herpesviridae Infections / Q000097:blood / Q000150:complications / Q000276:immunology / Q000382:microbiology*; D015654:Herpesvirus 6, Human / Q000235:genetics*; D006689:Hodgkin Disease / Q000150:complications; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000382:microbiology*; D016393:Lymphoma, B-Cell / Q000150:complications; D008297:Male; D008875:Middle Aged; D009103:Multiple Sclerosis / Q000150:complications; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012463:Saliva / Q000382:microbiology
D000914:Antibodies, Viral; D004279:DNA, Viral