MIP-1 and are secreted by isolated myeloma cells
from most of patients with multiple osteolytic lesions
105
104
ys)ad2
103
for
cells6
102
g/10
(p
10
MIP-1
MIP-1
MIP-1
MIP-1
bone lesions
< 2
2 2
MIP-1 and secreted by isolated myeloma cells
from myloma patients correlated with urinary Dpd
urinary
urinary
Abe M, et al. Blood 100: 2195, 2002Hashimoto T, et al. Br J Haematol 125: 38, 2004
Multiple myeloma and dendritic cells
1.
Multiple myeloma (MM) cells cause devastating bone
destruction by enhanced osteoclastogenesis.
2.
The number and functions of dendritic cells (DCs) are
reduced in MM, leading to suppression of anti-tumor
immunity and susceptibility to infection.
3.
Because osteoclasts (OCs) and myeloid DCs are
derived from the same monocytic precursor cells, we
hypothesized that MM cells may reciprocally regulate
differentiation processes of these two cell lineages.
Objectives
To clarify roles for MM cells and MIP-1
in OC and DC differentiation from
monocytes.
OC differentiation from human peripheral
blood monocytes
M-CSF (500 U/mL)
sRANKL (50 ng/mL)
Monocyte
OC
14-21 days
RANK+
TRAP-positive
add
multinucleated cells
Control IgG or
anti-MIP-1, (20 µg/mL)
Anti-MIP-1 and antibodies in combination
inhibited OC formation from human peripheral
blood monocytes
TRAP staining
RT-PCR
anti-
Control IgG MIP-1,
Control IgG
RANK
mRNA
GAPDH
anti-
MIP-1,
Exogenous MIP-1 dose-dependently induced
RANK mRNA expression in HL-60 monocytic cells
(ng/ml)
RNA protection assay
Induction of myeloid DCs from human peripheral
blood monocytes
GM-CSF (500 U/mL)
Further addition of
TNF- (25 ng/mL)
IL-4 (50 ng/mL)
7 days
1 day
CD14+ monocytes
immature DCs
mature DCs
CD1a+
CD1a+
-
CD83
CD83+
CD80+
CD80++
HLA class II+
HLA class II++
DC differentiation from monocytes was inhibited
by non-adhesive co-cultures with MM cells
Lymphocytes
Primary
(control)
RPMI8226
U266
MM cells
CD83
CD80
HLA
class II
Anti-MIP-1, restored mature DC formation
suppressed by MM cells
CD83 expression
IgG
anti-MIP-1,
anti-IL-6
anti-VEGF
Anti-MIP-1, restored immature DC formation
suppressed by MM cells
CD1a expression
IgG
anti-MIP-1,
anti-IL-6
anti-VEGF
DC function was determined by IFN-
production from T cells
MIP-1
15Gy
anti-CD3
or
(HIT3a)
RPMI8226
IFN-
+
production
GM-CSF
(ELISA)
CD14+
IL-4
DC
CD8+ T cell
monocyte
TNF-
DC functions are defective in the presence of
MIP-1 or RPMI 8226 cells
700
600
(pg/ml)
500
IFN-
400
300
200
100
T cell
None
MIP-1
RPMI
only
DC
OC and DC differentiation from monocytes in MM
RANK
pre OCs
OCs
Bone Resorption
RANKL
M-CSF
monocytes
MIP-1,
MM cells
Ag Presentation
GM-CSF
IL-4
mature
immature DCs
DCs