Clinical-grade functional dendritic cells from patients with multiple myeloma are not infected with Kaposi's sarcoma-associated herpesvirus.
Bone marrow dendritic cells (DC) from patients with multiple myeloma (MM) were recently reported to be infected with Kaposi's sarcoma-associated herpesvirus (KSHV). Because immunotherapy strategies using DC are very promising in this disease, we looked for KSHV DNA in clinical-grade DC generated in vitro from MM patients. Adherent apheresis cells from MM patients were maintained for 7 days in clinical-grade X-VIVO 15 culture medium supplemented with granulocyte-macrophage colony-stimulating factor, interleukin-4, or interleukin-13. Tumor necrosis factor alpha was added for the last 2 days. We obtained a cell population with a DC phenotype able to endocytose fluorescein isothiocyanate (FITC)-dextran and efficiently activate resting allogenic T lymphocytes. To detect KSHV DNA, we used polymerase chain reaction (PCR) followed by Southern blotting of PCR product with a sensitivity detecting a few copies of viral DNA. All the PCR were repeated in a blinded fashion three times, on 1 mug and 0.2 mug of genomic DNA, in two different laboratories. Clinical-grade DC from 10 (91%) of 11 patients were not infected with KSHV. The apheresis cells and the purified CD34(+) cells from the same patients were also negative. A very weak PCR band was detected with DC from one patient, but the initial apheresis cells were negative. The detection of KSHV infection in 1 (9%) of 11 MM patients probably represents background seroprevalence. It seems likely that functional and clinical-grade DC from MM patients can safely be used in clinical trials.
Tarte
K
K
Institute for Molecular Genetics, CNRS, Montpellier, France.
Olsen
S J
SJ
Yang Lu
Z
Z
Legouffe
E
E
Rossi
J F
JF
Chang
Y
Y
Klein
B
B
D001781:Blood Component Removal; D015139:Blotting, Southern; D001853:Bone Marrow / Q000473:pathology / Q000821:virology; D002469:Cell Separation; D002478:Cells, Cultured; D015897:Comorbidity; D004273:DNA, Neoplasm / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis; D003713:Dendritic Cells / Q000187:drug effects / Q000821:virology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000821:virology; D006566:Herpesviridae Infections / Q000453:epidemiology; D019288:Herpesvirus 8, Human / Q000302:isolation & purification*; D006801:Humans; D018793:Interleukin-13 / Q000494:pharmacology; D015847:Interleukin-4 / Q000494:pharmacology; D008875:Middle Aged; D009101:Multiple Myeloma / Q000453:epidemiology / Q000473:pathology / Q000821:virology*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D016037:Single-Blind Method
D004273:DNA, Neoplasm; D004279:DNA, Viral; D018793:Interleukin-13; D015847:Interleukin-4; D016178:Granulocyte-Macrophage Colony-Stimulating Factor