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Bone marrow derived dendritic cells from patients with multiple myeloma cultured with three distinct protocols do not
F Bellos1, H Goldschmidt, M Dörner
1Department of Internal Medicine V, University of Heidelberg, Germany.
Background:
An association between Kaposi's sarcoma associated herpesvirus (KSHV) and the pathogenesis of multiple myeloma (MM) was postulated recently. The dendritic cells of patients with MM were proposed to be infected with the virus.
Patients And Methods:
Bone marrow mononuclear cells (MNC) of 23 patients, 22 with MM and one with MGUS, were cultured according to three distinct protocols for the generation of dendritic cells. One was essentially the stromal 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 330233 sequence of the virus genome followed by hybridisation with a KSHV specific oligonucleotide.
Results:
FACS analysis of the cells with the specific markers CD1a, CD86 and HLA-DR, characteristic for dendritic cells, revealed differences in the expression pattern depending on the protocol used. The proportion of CD1a+ cells was very low in the stromal cell cultures (median 0.4%), while a higher 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 with 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 specific after hybridisation.
Conclusion:
We have no evidence that bone marrow derived dendritic cells from patients with MM are infected with KSHV.
Insights
This study investigated Kaposi's sarcoma associated herpesvirus (KSHV) in multiple myeloma (MM) patients. Researchers found no evidence of KSHV infection in bone marrow-derived dendritic cells from MM patients.
Area of Science:
- Hematology
- Virology
- Immunology
Background:
- A potential link between Kaposi's sarcoma associated herpesvirus (KSHV) and multiple myeloma (MM) pathogenesis has been proposed.
- Dendritic cells in MM patients were hypothesized to harbor KSHV infections.
Purpose of the Study:
- To investigate the presence of KSHV in dendritic cells derived from the bone marrow of multiple myeloma patients.
Main Methods:
- Bone marrow mononuclear cells from 23 patients (22 MM, 1 MGUS) were cultured using three protocols to generate dendritic cells.
- Cells were analyzed by FACS for dendritic cell markers and by nested PCR for KSHV DNA, followed by hybridization.
Main Results:
- FACS analysis showed varying proportions of dendritic cell markers (CD1a, CD86, HLA-DR) depending on the culture protocol.
- PCR assays did not detect KSHV DNA in any of the 34 assessed samples.
- Tentative PCR products were not confirmed as KSHV-specific upon hybridization.
Conclusions:
- The study found no evidence to support KSHV infection in bone marrow-derived dendritic cells from multiple myeloma patients.