Bone marrow and peripheral blood dendritic cells from patients with multiple myeloma are phenotypically and functionally normal despite the detectionof Kaposi's sarcoma herpesvirus gene sequences

Citation
N. Raje et al., Bone marrow and peripheral blood dendritic cells from patients with multiple myeloma are phenotypically and functionally normal despite the detectionof Kaposi's sarcoma herpesvirus gene sequences, BLOOD, 93(5), 1999, pp. 1487-1495
Citations number
56
Categorie Soggetti
Hematology,"Cardiovascular & Hematology Research
Journal title
BLOOD
ISSN journal
00064971 → ACNP
Volume
93
Issue
5
Year of publication
1999
Pages
1487 - 1495
Database
ISI
SICI code
0006-4971(19990301)93:5<1487:BMAPBD>2.0.ZU;2-E
Abstract
Multiple myeloma (MM) cells express idiotypic proteins and other tumor-asso ciated antigens which make them ideal targets for novel immunotherapeutic a pproaches. However, recent reports show the presence of Kaposi's sarcoma he rpesvirus (KSHV) gene sequences in bone marrow dendritic cells (BMDCs) in M M, raising concerns regarding their antigen-presenting cell (APC) function. In the present study, we sought to identity the ideal source of DCs from M M patients for use in vaccination approaches. We compared the relative freq uency, phenotype, and function of BMDCs or peripheral blood dendritic cells (PBDCs) from MM patients versus normal donors. DCs were derived by culture of mononuclear cells in the presence of granulocyte-macrophage colony-stim ulating factor and interleukin-4. The yield as well as the pattern and inte nsity of Ag (HLA-DR, CD40, CD54, CD80, and CD86) expression were equivalent on DCs from BM or PB of MM patients versus normal donors. Comparison of PB DCs versus BMDCs showed higher surface expression of HLA-DR (P = .01), CD86 (P = .0003), and CD14 (P = .04) on PBDCs. APC function, assessed using an allogeneic mixed lymphocyte reaction (MLR), demonstrated equivalent T-cell proliferation triggered by MM versus normal DCs. Moreover, no differences i n APC function were noted in BMDCs compared with PBDCs. polymerase chain re action (PCR) analysis of genomic DNA from both MM patient and normal donor DCs for the 233-bp KSHV gene sequence (KS330(233)) was negative, but nested PCR to yield a final product of 186 bp internal to KS330(233) was positive in 16 of 18 (88.8%) MM BMDCs, 3 of 8 (37.5%) normal BMDCs, 1 of 5 (20%) MM PBDCs, and 2 of 6 (33.3%) normal donor PBDCs. Sequencing of 4 MM patient P CR products showed 96% to 98% homology to the published KSHV gene sequence, with patient specific mutations ruling out PCR artifacts or contamination. In addition, KHSV-specific viral cyclin D (open reading frame [ORF] 72) wa s amplified in 2 of 5 MM BMDCs, with sequencing of the ORF 72 amplicon reve aling 91% and 92% homology to the KSHV viral cyclin a sequence. These seque nces again demonstrated patient specific mutations, ruling out contaminatio n. Therefore, our studies show that PB appears to be the preferred source o f DCs for use in vaccination strategies due to the ready accessibility and phenotypic profile of PBDCs, as well as the comparable APC function and low er detection rate of KSHV gene sequences compared with BMDCs. Whether activ e KSHV infection is present and important in the pathophysiology of MM rema ins unclear; however our study shows that MMDCs remain functional despite t he detection of KSHV gene sequences. (C) 1999 by The American Society of He matology.