Haematologica 2003 - Supplements
Haematologica 2003 - Supplements
Haematologica 2003 - Supplements
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1/MAGE-A3 immunotherapy can be used as secondary<br />
prevention to prevent relapse/disease progression in NY-ESO-<br />
1/MAGE-A3 negative patients receiving chemotherapy or in<br />
smoldering/indolent myeloma.<br />
403<br />
Dendritic cell numbers and their subsets during<br />
treatment of multiple myeloma<br />
1KOVAROVA, Lucie, MSc; 1,2HAJEK, Roman, MD;<br />
3SVOBODNIK, Adam, MSc; 1KLABUSAY, Martin, MD,<br />
PhD; 1VORLICEK, Jiri, Professor, MD<br />
1Department of Internal Medicine-Hematooncology, Masaryk<br />
University Hospital Brno-Bohunice, Jihlavska 20, 625 00 Brno,<br />
Czech Republic2Department of Clinical Hematology, Masaryk<br />
University Hospital Brno-Bohunice, Jihlavska 20, 625 00 Brno,<br />
Czech Republic3Centre of Biostatistics and Analysis, Masaryk<br />
University Brno-Bohunice, Kamenice 126/3,625 00 Brno<br />
Background and objective: In this study, the proportion of<br />
dendritic cell (DC) subsets (DC1, DC2) in peripheral blood of<br />
patients with multiple myeloma (MM) was evaluated before and<br />
during treatment.<br />
Design and Methods: Flow cytometric determination of DC<br />
subsets in peripheral blood was based on positive expression of<br />
the surface antigen CD83 in combination with HLA-DR and<br />
either CD11c or CD123.<br />
Results: No significant differencies were found in initial values<br />
between the group of healthy volunteers (n = 15; mean count of<br />
CD83+ cells was 0.62±0.06%; ratio DC1/DC2 = 2.5) and the<br />
group of patients before treatment (n = 15; 0.59±0.13% CD83+;<br />
DC1/DC2 = 2.17). In a group of patients (n = 10) treated with<br />
VAD regimen (vincristine, adriamycin, dexamethasone), the<br />
mean percentage of DC after induction treatment (0.84±0.4%<br />
CD83+ cells; DC1/DC2 = 1.59) was higher than initial values.<br />
Administration of G-CSF reduced the total DC numbers<br />
(0.66±0.6%; DC1/DC2 = 3.71). The lowest total DC counts were<br />
in the apheresis products (0.36±0.2%; DC1/DC2 = 4.75).<br />
Administration of GM-CSF increased DC numbers (0.56±0.3%;<br />
DC1/DC2 = 1.59). Pretreatment DC values were achieved within<br />
six months after transplantation (0.83±0.6%; DC1/DC2 = 3.92).<br />
Interpretation and Conclusions: The highest number of total DC<br />
was found after induction treatment and within six months of<br />
transplantation. The ratio DC1/DC2 showed a the highest value in<br />
the apheresis products and in peripheral blood within the first six<br />
months after transplantation.<br />
404<br />
IL-12 corrects the dendritic cell defect of patients with<br />
myeloma<br />
Ross Brown1, Allan Murray1, Belinda Pope1, Daniel M<br />
Sze1, John Gibson1, P Joy Ho1, Derek Hart2, Doug<br />
Joshua1<br />
1Institute of Haematology, Royal Prince Alfred Hospital, Sydney;<br />
2Mater Medical Research Institute, Brisbane, Australia.<br />
The poor response to immunotherapy in patients with multiple<br />
myeloma indicates that a better understanding of the immune<br />
defects in this disease is required before more effective<br />
therapeutic strategies can be developed. Recently we reported<br />
that high potency (CMRF44+) dendritic cells (DC) in the<br />
peripheral blood of patients with multiple myeloma failed to<br />
upregulate the expression of the B7 costimulatory molecules,<br />
CD80 and CD86, in response to an appropriate signal from<br />
huCD40LT. During antigen presentation to T cells the level of<br />
expression of the B7 molecules provides an important “second<br />
signal” that determines the fate of each cell – apoptosis, anergy or<br />
productive immunity. We have demonstrated that this defect is<br />
caused by TGF1 and IL-10 produced by malignant plasma cells<br />
and that it is possible to neutralise the defect in vitro with anti-<br />
TGF1. If this defect has an impact on immunotherapy<br />
strategies, it would be important to identify a more suitable agent<br />
than anti-TGF1 for in vivo use. The number of high potency<br />
DC (CMRF44+, CD14-, CD19-, PI-) in the blood of patients with<br />
myeloma (0.03-0.8% of mononuclear cells; n= 26) was not<br />
significantly different from normal controls (0.05-0.8% of<br />
MNCs; n=13). The expression of the costimulatory molecules<br />
CD80 and CD86 on blood DC of these patients (2917% and<br />
8510% of MNCs respectively) was also normal (2917% and<br />
8616% of MNCs). Incubation with huCD40LT stimulated<br />
upregulation of CD80 expression on the DC and B cells of<br />
normal controls but there was either reduced or no upregulation<br />
of CD80 on the DC of the patients with myeloma. Less than 10%<br />
of malignant plasma cells expressed CD80 and huCD40LT failed<br />
to significantly upregulate CD80 expression on mature plasma<br />
cells (n=6). Upregulation of CD80 on DC of normal controls was<br />
inhibited by rTGF-in a dose dependent manner. CD86<br />
expression on DC was high both before (86%) and after (89%)<br />
stimulation. IL-12 but not interferon-could replace anti-TGF1<br />
as an agent capable of neutralising the failure to stimulate CD80<br />
upregulation by huCD40LT. DC subsets stimulated by IL-12<br />
were predominantly myeloid (CD11c+ and CDw123- ),<br />
suggesting that they could initiate a Th1 type response. Thus<br />
patients with myeloma have a normal number of DC but may fail<br />
to upregulate CD80 expression in the presence of huCD40LT due<br />
to tumour-derived TGF-and/or IL-10. This defect can be<br />
corrected with IL-12.<br />
405<br />
IN VITRO STIMULATION OF LYMPHOCYTES FROM<br />
MULTIPLE MYELOMA PATIENTS WITH AUTOLOGOUS<br />
DENDRITIC CELLS PULSED WITH PLASMOCYTE<br />
LYSATES.<br />
L. Garderet, C. Mazurier, C. Funck-Brentano, A. Karim, S.<br />
Bouchet, J. Frick, NC. Gorin, M. Lopez.<br />
Department of Hematology, CIC, EFS, LTCRA, Inserm U 76-CHU<br />
Saint Antoine, 27 rue de Chaligny- 75012, Paris; 92-Fontenay Aux<br />
Roses, France.<br />
Tumor vaccination using dendritic cells (DCs) pulsed with tumor<br />
antigens has proven useful mainly in melanomas, prostate<br />
cancers, and lymphomas.Experimental data in mouse models or<br />
in in vitro human models have established that multiple myeloma<br />
(MM) is immunogenic and can induce an immune T response<br />
when the tumor antigen is adequately presented to T lymphocytes<br />
by DCs. In a pre-clinical study, we are evaluating the possibility<br />
to generate in MM cytotoxic T lymphocytes directed against the<br />
autologous patient’s tumor cells, i.e. the plasmocytes in a<br />
protocol that fits GMP procedures. Until now, 8 patients (pts)<br />
have been collected. Plasmocytes were isolated from bone<br />
marrow at diagnosis (except for 2 pts, for whom the monoclonal<br />
immunoglobulin was used as tumor antigen), selected with anti-<br />
CD138 magnetic beads and cryopreserved. Monocytes and<br />
lymphocytes were purified by elutriation from apheresis products<br />
of pts at remission . Monocytes were differentiated into DCs by<br />
culture during 5 days in teflon bags in the presence of GM-<br />
CSF+IL-13+fetal calf serum. DCs were pulsed with autologous<br />
plasmocyte lysates or monoclonal Ig, matured using poly-IC +<br />
anti-CD40, and then cocultured with autologous T lymphocytes.<br />
The cytotoxicity of T lymphocytes against autologous<br />
plasmocytes (or Ig-pulsed DCs) was tested using a 51chromiumrelease<br />
assay. The production of interferon-γ by stimulated<br />
S268