Controversy exists as to whether Kaposi's sarcoma-associated herpesvirus (KSHV) is more widespread than originally reported. Recently, Monini et al reported that KSHV is ubiquitous in urogenital and prostate tissues and sperm of healthy Italian adults using nested polymerase chain reaction (PCR). We have examined for the presence of KSHV in 10 normal prostates from Italian men and 10 from men from the United States, as well as 32 prostatic, 30 vulvar, 24 ovarian, 20 cervical, and 30 testicular cancer specimens from patients from the United States. None of the patients had a history of human immunodeficiency virus infection. The samples were tested by nested PCR. The sensitivity of this assay was determined by a dilution study performed by diluting KSHV DNA from the KS-1 cells (a primary effusion lymphoma cell line which is estimated to have 16 copies of KSHV per cell) in DNA from a K562 myeloid cell line. The nested PCR that we used can detect 2.4 copies of KSHV sequences on a background of K562 DNA. All the samples were negative for KSHV sequences. Therefore, we cannot confirm the finding that KSHV sequences are ubiquitous in urogenital and prostate tissues. Furthermore, because our samples were from both the United States and Italy, the discrepancy between results is unlikely to be explained by either ethnic or environmental factors. False-positive results easily occur using nested primer PCR because of contamination. Our data argue that KSHV is not widely disseminated in urogenital tissues from nonimmunosuppressed individuals.
"This has been used successfully to confirm the presence of KSHV DNA in KS, PEL or MCD tissue samples, and in PBMC or saliva from immunosuppressed AIDS or KS patients (Moore and Chang,1995; Cesarman,1995; Whitby,1995; Soulier,1995; Marchioli, 1996; Huang,1995). However, some other clinical or epidemiological studies have claimed detection of KSHV DNA by PCR in other disease conditions, such as multiple myeloma (MM), sarcoid, primary pulmonary hypertension, pemphigus, Bowen's disease, or Kikuchi disease, and in saliva or urogenital specimens from seronegative patients (Luppi,1997; Chauhan, 1999; Hsu,2001; Gao,1999; Said,1997a; Tisdale,1998; Monini,1996; Rettig,1997; Di Alberti, 1997b; Di Alberti,1997a; Cool,2003; Agbalika,1998; Brousset,2000; Boralevi,1998; Said, 1996; Kikuta,1997; Tasaka,1997; Brousset,1997; Huh,1998; Sjak-Shie,1999; Berenson and Vescio,1999; Nishimoto,1997; Memar,1997; Merelli,1997; Said,1997b). For the most part those claims have not been reproducible by other investigators, or by other techniques, and have not gained wide acceptance (Pan,2001; Tarte,1999; Moore and Chang,1998; Moore, 1998; Henke-Gendo,2005; Corbellino,1999). "
[Show abstract][Hide abstract] ABSTRACT: Ever since the original identification of fragments of KSHV DNA in Kaposi's sarcoma (KS) tissue by Chang et al. in 1994, PCR has been used successfully and extensively to detect the virus in clinical samples from the accepted etiological diseases of KS, PEL and MCD. However, a number of other clinical and epidemiological studies claiming evidence for KSHV in multiple myeloma or sarcoid and more recently in primary pulmonary hypertension, as well as claims about the biological significance of DNA sequence polymorphisms based just on small ORF26 PCR DNA fragments have not been convincing. Here, we evaluate the validity and interpretations of previous results in the context of both the observed rates and global patterns of sequence variability within an extended ORF26 locus, as well as from the perspective of the overall levels of KSHV variability found after sampling multiple loci across the complete KSHV genome. The results cast doubts on most claims for biological significance for these polymorphisms, which instead correlate with viral subtype clustering arising from geographic and ethnic divergence of the ancestral human hosts. In addition, we describe several observations that help to explain likely sources of the often either unexpectedly high or unexpectedly low levels of sporadic variability seen in the PCR DNA sequence data reported in some of those studies.
[Show abstract][Hide abstract] ABSTRACT: The aim of our study was to test if dendritic cells contain the KSHV genome. CD34+ peripheral blood progenitor cells (PBPC) and bone marrow mononuclear cells were cultured in X-VIVO 15 medium supplemented with GM-CSF and TNF-alpha in gas-permeable containers. Dendritic cells were identified morphologically and immunophenotypically. The KSHV genome was not identified in any of the cases using a nested primer PCR approach. Serological analysis corroborated the molecular findings: no antibodies for KSHV were found in any of the multiple myeloma patients. These data are of importance when considering use of DC for therapeutic approaches in multiple myeloma.
British Journal of Haematology 10/1998; 102(5):1338-40. DOI:10.1046/j.1365-2141.1998.00894.x · 4.71 Impact Factor
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