Abstract Introduction Experiments ans Results Conclusions

Memory phenotype CD8

+
T cells are superior to naive CD8
+

T cells in separating graft anti-tumor activity from GVHD after
bone marrow transplantation: Application To DLI
Division of Immunology and Rheumatology
1
, Blood and Marrow Transplant
2
, Hematology and Oncology
4
, Departments of Medicine and Pathology
3
, Stanford University School of Medicine, Stanford , California, USA
We compared the graft anti-tumor effect and GVHD
activities of naturally occurring naive and memory
phenotype subsets of C57BL/6 (H-2
b
) CD8
+
T cells after
bone marrow transplantation into BALB/c (H-2
d
) mice.
The tumor used for the study was the BCL
1
lymphoma, a
spontaneously arising non-transfected cell line. We found
that naive CD8
+
T cells (CD62L
hi
CD44
lo
) express higher
levels of gut homing molecules
4

7
, CCR9 and CD103
compared to memory subsets comprised of both effector
(CD62L
lo
CD44
hi
) and central memory cells
(CD62L
hi
CD44
hi
). The proliferation of the naive cells was
ten-fold higher than memory phenotype cells against
BALB/c stimulators and memory cells secreted
significantly more IFN in the MLR. All BALB/c hosts
without tumor cells that were given donor memory CD8
+

T cells along with T cell depleted bone marrow cells
(TCD BM) survived over 100 days of transplantation
without significant weight loss compared to TCDBM
controls. However, 25% of the mice given naive CD8
+
T
cells and TCD BM died due to GVHD and had
significantly greater weight loss compared to TCD BM
controls or to hosts given memory CD8
+
T cells and TCD
BM. BALB/c hosts given 500 BCL
1
and TCD BM all
died of lymphoma within 28 days of transplantation.
Hosts given BCL
1
cells, TCD BM and either memory or
naive cells cleared the tumor by day 28 but weight loss
and survival was significantly improved in the memory
versus naive cell groups despite non significant
differences in the GVHD scores in the liver and gut at
day 100. Ex vivo imaging of mice receiving BCL
1
tumor
cells, TCD BM, and CD8
+
T cells from luciferase
transgenic donors showed increased accumulation of
naive phenotype CD8
+
T cells in the liver and the gut
compared to memory phenotype cells after 6 days of
transplantation. We compared the memory and naive
CD8
+
T cells in a model of donor lymphocyte infusion
(DLI) in which BALB/c mice with progressive growth of
BCL
1
tumor after TCD BM transplantation were given
infusion of the CD8
+
T cells. Both naive and memory
cells are effective in clearing the tumor, and converting
the hosts from mixed to complete chimeras. Differences
in GVHD are observed. In conclusion, CD8
+
memory
phenotype T cells are a desirable subset that mediates
potent anti-tumor activity without severe GVHD, and can
be used in a DLI model.
In a MHC mismatch model of BMT, donor
CD8
+
T cells mediate anti-tumor effect
against BCL
1
lymphoma. (Bone Marrow
Transplant 2007, 40: 487; Transplantation
1995, 60: 355). However, these cells also
induce lethal GVHD.

Unprimed CD62L
-
memory T cells were
unable to induce GVHD, whereas, BCL
1

tumor primed CD62L
-
memory T cells
inhibited the lymphoma growth. (Blood
2004, 103:1534).
It is not clear whether unprimed memory
phenotype CD8
+
T cells can mediate Graft
versus Lymphoma effect without inducing
lethal GVHD.
Donor lymphocyte infusions are used to
treat lymphoma or leukemia relapse in
patients after allogeneic bone marrow
transplantation, however, these donor
lymphocytes also induce lethal GVHD.
Therefore, we addressed the use of naive
and memory phenotype CD8
+
T cells as
donor lymphocyte infusions for potential
GVL effect sparing lethal GVHD.
Experiments and Results
Naive CD8
+
T cells express high levels of gut
homing molecules
Suparna Dutt
1
, Jeanette Baker
2
, Neeraja Kambham
3
, Holbrook E. Kohrt
4
, Robert S. Negrin
2
and Samuel Strober
1
0102 103 104 105 0102 103 104 105
CCR9
0102 103 104 105
CD103
0
20
40
60
80
100
47
010 2 10 3 10 4 10 5
CD44
0
10 2
10 3
10 4
10 5
20
40
60
80
100
010 2 10 3 10 4 10 5
0
10 2
10 3
10 4
10 5
18.2
79.4
Naive
Memory
Allo Naive Allo Memory Syn Naive Syn Memory
0
50000
100000
150000
200000
Naive CD8
+
T cells proliferate more than
memory phenotype CD8
+
T cells in MLR
Allo Naive Allo Memory Syn Naive Syn Memory
0
250
500
750
1000
1250
1500
1750
2000
Memory phenotype CD8
+
T cells produce more
IFN than naive CD8
+
T cells
Naive CD8
+
T cells produce more IL-2 than
memory phenotype CD8
+
T cells
Allo Naive Allo Memory Syn Naive Syn Memory
0
50
100
150
200
IFN
IL-2
0 25 50 75 100 125
0
25
50
75
100
125
TCDBM + Naive CD8+ T cells
TCD BM + Memory CD8+ T cells
TCDBM
Days after transplantation
0 25 50 75 100 125
0
5
10
15
20
25
30
35
TCDBM + Naive CD8+ Tcells
TCD BM + Memory CD8+ T cells
TCD BM
Days after transplantation
Memory phenotype CD8
+
T cells do not induce
lethal GVHD
Memory phenotype CD8
+
T cells mediate GVL
without inducing lethal GVHD
0 25 50 75 100 125
0
25
50
75
100
125
TCD BM+ Naive CD8+T cells
TCD BM+ Memory CD8+ T cells
TCDBM BCLI only
Days after transplantation
0 25 50 75 100 125
0
5
10
15
20
25
30
TCDBM + Naive CD8+ Tcells
TCD BM + Memory CD8+ T cells
TCD BM
Days after transplantation
No significant differences in histopathology scores between
Naive and Memory phenotype CD8
+
T cells groups
Liver Colon Small Bowel
0.0
0.5
1.0
1.5
2.0
2.5
TCD BM+ Naive CD8+ T cells
TCD BM+Memory CD8+ T cells
TCD BM
* * * * signficant differences(p<0.05)
between the Naive and TCD
BM group
Experimental scheme for DLI
Characterization of naive and
memory phenotype CD8
+
T cells
Experimental design for bone
marrow transplantation
Naive phenotype CD8
+
T cells accumulate more
in the gut than memory phenotype CD8
+
T cells
Naive and memory phenotype CD8
+
T cells
equally clear BCL
1
lymphoma
0 20 40 60 80
0
20
40
60
80
100
No DLI
DLI Naive CD8 + T
DLI Memory CD8 + T
Days after transplantation of TCD BM
Naive
Memory
No DLI
Memory phenotype CD8
+
T cells as DLI clear BCL
1

lymphoma cells without inducing lethal GVHD
This work was supported by a grant from
the Leukemia and Lymphoma Society.
There are no relevant conflicts of interest
to disclose.
Memory phenotype CD8
+
T cells
proliferate significantly lower than
naive CD8
+
T cells in MLR.
They produce higher levels of IFN
and lower levels of IL-2 than naive
CD8
+
T cells cells.
Memory phenotype CD8
+
T cells do
not induce lethal GVHD.
Memory phenotype CD8
+
T cells
mediate Graft versus Lymphoma
effect against BCL
1
lymphoma cells.
Memory phenotype CD8
+
T cells show
decreased accumulation in the gut
compared to naive CD8
+
T cells.
As donor lymphocyte infusions,
memory phenotype CD8
+
T cells clear
BCL
1
lymphoma without inducing
lethal GVHD.