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Targeted integration of human herpesvirus 6 in the p arm of chromosome 17 of human peripheral blood mononuclear cells in vivo
Author(s) -
Torelli Giuseppe,
Barozzi Patrizia,
Marasca Roberto,
Cocconcelli Paola,
Merelli Elisa,
CeccheriniNelli Luca,
Ferrari Sergio,
Luppi Mario
Publication year - 1995
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.1890460303
Subject(s) - peripheral blood mononuclear cell , virology , in vivo , peripheral blood , human herpesvirus , chromosome , biology , virus , peripheral , in vitro , immunology , medicine , gene , genetics
Out of 64 cases of non‐Hodgkin's lymphomas (NHL), 55 cases of Hodgkin's disease (HD) and 31 cases of multiple sclerosis (MS), 2 NHL, 7 HD and 1 MS cases were found positive by polymerase chain reaction (PCR) for the presence of HHV‐6 sequences in pathologic lymph nodes of the lymphomas and in peripheral blood mononuclear cells (PBMCs) of MS. A further analysis of the PBMCs of the PCR positive cases by standard Southern blot technique revealed only 2 NHL, 3 HD and 1 MS cases as positive, indicating that these six patients have an unusually high viral copy number in the PBMCs. Restriction analysis, carried out using probes representative of differ ent regions of the virus, showed that three cases retain only a deleted portion of the viral genome. In the remaining three cases a complete viral genome was present, containing the right end sequences in which the rep‐like gene, possibly crucial to the viral and cellular life cycle, is lo cated. The analysis by pulsed field gel electrophoresis (PFGE) of the total DNA of the PBMCs obtained directly, without culture from PBMCs of these last three cases (1 NHL, 1 HD, and 1 MS), using the same probes, showed the absence of free viral molecules and the association of viral sequences with high molecular weight DNA. These results are consistent with in vivo integration of the entire virus in the cellular genome. A further study of the same patients with chromo some fluorescence in situ hybridization (FISH) showed in ail the three cases the presence of a specific hybridization site, located at the telomeric extremity of the short arm of chromosome 17 (17p13), suggesting that this location is at least a preferred site of an infrequent, but possi bly biologically important, integration phenomenon. © 1995 Wiley‐Liss, Inc.