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Clinical-Grade Functional Dendritic Cells From Patients With Multiple
Myeloma Are Not Infected With Kaposi's Sarcoma-Associated Herpesvirus
Karin Tarte,
Sonja J. Olsen,
Zhao Yang Lu,
Eric Legouffe,
Jean-François Rossi,
Yuan Chang, and
Bernard Klein
From the Institute for Molecular Genetics, CNRS, Montpellier, France;
the Division of Epidemiology, School of Public Health, Columbia
University, New York, NY; the Unit for Cellular Therapy, CHU
Montpellier, Hopital Saint Eloi, Montpellier, France; the Service des
Maladies du Sang B, CHU Montpellier, Hopital Lapeyronie, Montpellier,
France; and the Department of Pathology, College of Physicians and
Surgeons, Columbia University, New York, NY.
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  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 µg and 0.2 µg 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.
Blood, Vol. 91 No. 6 (March 15), 1998:
pp. 1852-1857
© 1998 by The American Society of Hematology.
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