Bone marrow derived dendritic cells from patients with multiple myeloma cultured with three distinct protocols do not bear Kaposi's sarcoma associated herpesvirus DNA

Citation
F. Bellos et al., Bone marrow derived dendritic cells from patients with multiple myeloma cultured with three distinct protocols do not bear Kaposi's sarcoma associated herpesvirus DNA, ANN ONCOL, 10(3), 1999, pp. 323-327
Citations number
25
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Oncology,"Onconogenesis & Cancer Research
Journal title
ANNALS OF ONCOLOGY
ISSN journal
0923-7534 → ACNP
Volume
10
Issue
3
Year of publication
1999
Pages
323 - 327
Database
ISI
SICI code
0923-7534(199903)10:3<323:BMDDCF>2.0.ZU;2-7
Abstract
Background: An association between Kaposi's sarcoma associated herpesvirus (KSHV) and the pathogenesis of multiple myeloma (MM) was postulated recentl y. The dendritic cells of patients with MM were proposed to be infected wit h the virus. Patients and methods: Bone marrow mononuclear cells (MNC) of 23 patients, 2 2 with MM and one with MGUS, were cultured according to three distinct prot ocols for the generation of dendritic cells. One was essentially the stroma l cell culture protocol described by Rettig et al. (Science 1997; 276: 1851 -4), while the two other protocols comprised growth factors. Cultured cells were characterised by FACS analysis and assessed for the presence of KSHV DNA with a highly sensitive and specific nested PCR assay detecting the KS 330(233) sequence of the virus genome followed by hybridisation with a KSHV specific oligonucleotide. Results: FAGS analysis of the cells with the specific markers CD1a, CD86 an d HLA-DR, characteristic for dendritic cells, revealed differences in the e xpression pattern depending on the protocol used. The proportion of CD1a+ c ells was very low in the stromal cell cultures (median 0.4%), while a highe r percentage of CD14+ cells could be observed (median 37.8%). Growth factor containing cultures revealed a distinctly higher median percentage of CD1a + cells of 32.5%. The proportion of CD86+ cells varied between 10.4% and 78 .5% and HLA-DR+ cells between 26% and 94.4%. Examination of those cells wit h PCR did not reveal positivity for KSHV in any of the 34 samples assessed. Amplification of seven samples revealed PCR products of approximately the size of the KS 330(233), which, however, could not be confirmed as KSHV spe cific after hybridisation. Conclusion: We have no evidence that bone marrow derived dendritic cells fr om patients with MM are infected with KSHV.