Inst
Ins itut
u e of Immunology
Rikshosp
s italet Uni
n versi
s ty
Hosp
s ital
T Cells in Myeloma: Lessons
from a Murine Model
Bjarne Bogen
University of Oslo
http://immunol.net

Lynch....Eisen PNAS -72:
Vaccination with myeloma protein induces resistance
against tumor challenge with MM cells.
MOPC315 s.c.
BALB/c
Tumors
MOPC315 s.c.
M315/CFA s.c.
Protection
MOPC315 s.c.
M460/CFA s.c.
Tumors

V
V
V
H
L
H
VL
M315 IgA
M460 IgA
Idiotypes
Idiotype(Id)-vaccination

Id-vaccination:
What arms of anti-Id Immunity are effective?
Antibodies?
CD8+ T cells?
CD4+ T cells?

The PC315 Idiotype model
MOPC315
V1/V8.2
4B2A1
Balb/c CD4+
Th1 clone
91-101
2315
I-Ed
TCR/ genes
91-101
Id-specific TCR-
Id-peptide
91 92 93 94
95
96 97 98 99 100 101
transgenic
mouse
Germline 2 AL
W Tyr Ser Thr H
F
V F
G
Id-specific T cell
2315
- Phe Arg Asn
--
-
-
-
--
Bogen et al Eur J Immunol 1986 a,b
Bogen et al EMBO-J 1989
Bogen et al Eur J Immunol 1992
Weiss, Bogen PNAS 1989
Weiss, Bogen Cell 1991

Tumor challenge in TCR-transgenic mice
Non-TG mice
TCR-TG mice
MOPC 315
1.0
Survival
0.8
TCR-TG mice
0.6
0.4
non-TG littermates
0.2
0
4
8
12
16
20
Weeks
Lauritzen et al PNAS 1994
Bogen et al Eur.J.Immunol. 95

What happens when CD4+ T cells reject class
II negative multiple myeloma cells?
· Problem: Difficult to study since there is no specimen
· Solution: Inject MM cells in Matrigel (collagen-based gel)
Corthay et al Immunity 2005

MOPC315 plasmacytoma tumor cells embedded
in a subcutaneous collagen gel (Matrigel)

Tumor-specific CD4+ T cells become activated in draining LN
and migrate to the incipient tumor site

Tumor-specific CD4+ T cells become activated in draining LN
and migrate to the incipient tumor site
LN
LN
Matrigel
TCR-TG
Matrigel
with
SCID
with
J558
MOPC315

Tumor-specific CD4+ T cells become activated in draining LN
and migrate to the incipient tumor site
MOPC315-specific
CD4+ T cells in right LN
ountsC
100
101
102
103
104
LN
LN
CD69
Matrigel
TCR-TG
Matrigel
with
SCID
with
J558
MOPC315

Tumor-specific CD4+ T cells become activated in draining LN
and migrate to the incipient tumor site
MOPC315-specific
MOPC315-specific
CD4+ T cells in left LN
CD4+ T cells in right LN
ountsC
ountsC
100
101
102
103
104
100
101
102
103
104
CD69
LN
LN
CD69
Matrigel
TCR-TG
Matrigel
with
SCID
with
J558
MOPC315

Tumor-specific CD4+ T cells become activated in draining LN
and migrate to the incipient tumor site
MOPC-specific
MOPC-specific
CD4+ T cells in left LN
CD4+ T cells in right LN
CD11a
CD44
ountsC
ountsC
CD62L
100
101
102
103
104
100
101
102
103
104
CD69
LN
LN
CD69
Matrigel
TCR-TG
Matrigel
with
SCID
with
J558
MOPC315

Tumor-specific CD4+ T cells become activated in draining LN
and migrate to the incipient tumor site
MOPC315-specific
MOPC315-specific
CD4+ T cells in left LN
CD4+ T cells in right LN
CD11a
CD44
ountsC
ountsC
CD62L
100
101
102
103
104
100
101
102
103
104
CD69
LN
LN
CD69
Matrigel
TCR-TG
Matrigel
with
SCID
with
J558
MOPC315

MHC class II+ macrophages invade the matrigel
plug containing CD138+ myeloma cells
C
C
D

.....and come in contact with CD138+ tumor cells.
Macrophages pick up myeloma protein, process it and present
Id-peptides on their MHC class II molecules.
D

Tumor-specific CD4+ T cells arrive a few days later and contact
Id-primed macrophages in the matrigel plug.
Tumor-specific CD4+ T are of a Th1 phenotype, become activated, and
secrete IFN and TNF which activates macrophages

T cell-activated macrophages suppress tumor cell growth
)-3 120
100
(cpmx10
80
60
growth
40
20
MOPC315
0
MOPC
0.8:1
4:1
20:1
0.8:1
4:1
20:1
only
TCR-Transgenic SCID
SCID
Matrigel-infiltrating macrophages + MOPC315
(effector : target ratio)

The mechanism of cancer immunosurveillance
mediated by myeloma-specific CD4+ T cells
Tissue
MHC class II
Myeloma protein
Draining
negative
lymph node
MM cell
naïve
CD4

The mechanism of cancer immunosurveillance
mediated by myeloma-specific CD4+ T cells
Tissue
MHC class II
Myeloma protein
Draining
negative
lymph node
MM cell
DC
naïve
CD4

The mechanism of cancer immunosurveillance
mediated by myeloma-specific CD4+ T cells
Tissue
MHC class II
Myeloma protein
Draining
negative
lymph node
MM cell
DC
naïve
Primed TCR
CD4
CD4

The mechanism of cancer immunosurveillance
mediated by myeloma-specific CD4+ T cells
Tissue
MHC class II
Draining
Myeloma protein
negative
lymph node
MM cell
DC
Primed TCR
CD4
naïve
CD4

The mechanism of cancer immunosurveillance
mediated by myeloma-specific CD4+ T cells
Tissue
MHC class II
Draining
Myeloma protein
negative
lymph node
MM cell
DC
Primed TCR
CD4
naïve
CD4
MHC II
M

The mechanism of cancer immunosurveillance
mediated by myeloma-specific CD4+ T cells
Tissue
MHC class II
Draining
Myeloma protein
negative
lymph node
MM cell
DC
Primed TCR
CD4
naïve
CD4
MHC II
Id peptide
M

The mechanism of cancer immunosurveillance
mediated by myeloma-specific CD4+ T cells
Tissue
MHC class II
Draining
Myeloma protein
negative
lymph node
MM cell
DC
Primed
CD4
naïve
TCR
CD4
MHC II
Id peptide
M

The mechanism of cancer immunosurveillance
mediated by myeloma-specific CD4+ T cells
Tissue
MHC class II
Draining
Myeloma protein
negative
lymph node
MM cell
MM growth
DC
inhibition
Primed
CD4
naïve
TCR
CD4
MHC
MH II
Id peptide
IFN
M
M

Problem: MOPC315 is a mineral oil induced plasmacytoma that grows
s.c. or i.p. as extramedullary plasmacytomas.
Question: Does Id-specific CD4+ T cells protect against MM disease in
the bone marrow?
Solution:
· By repetitive i.v. injections and recovery from femurs of
paraplegic mice, a variant of MOPC315 was obtained that
homes to the bone marrow a model for MM bone
marrow disease
· This cell line was labeled so that it constitutively expresses
Luciferase. This approach enables imaging of in vivo
growth.

non-transgenic mouse
d.7
d.12
d.18
d.27
d.33
Ventral
Image
Min = -4.8779e+05
Max = 1.3472e+05
p/sec/cm^2/sr
60000
50000
Dorsal
40000
30000
20000
Left
10000
Color Bar
Min = 7500
Max = 60000
Right

MOPC315 derived luminescence from ventral side
9000
8000
non TCRtg
TCR tg
7000
6000
cm2
5000
/sec/
4000
ons
3000
phot
2000
1000
0
Day 7
Day 12
Day 16
Day 25
Conclusion: Id-specific CD4+ T cells protect against MM disease
in the bone marrow

Inst
Ins itut
u e of Immunology
Acknowledgements
Riksho
h sp
s italet Univer
v si
s ty
Hosp
s ital
· Contributors in Bogen lab,
Institute of Immunology, University of
Oslo:
Ludvig Munthe
Collaborators:
Alexandre Corthay
Katrin Lundin
Dag Skovseth and Guttorm Haraldsen,
Institute of Pathology, University of Oslo
Agnete B. Fredriksen
Marianne Frøyland
Karoline W. Schjetne
Tuva Hereng
Michael Zangani
Hege Pihlstrøm
Yueqiu-F Gao
Gro Tunheim
Egil Røsjø
Peter Hofgaard
Hilde Omholt
Mona Lindeberg

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