Ehrlich, R.

Fazilleau, N.

Firat, H.

Gluckman, E.

Lemonnier, F.

Loiseau, P.

Rohrlich, P.S.

Elsevier - Publisher Connector 

T cells would ameliorate the development of graft-versus-host-
disease (GVHD) in recipients of an allogeneic bone marrow trans-
plantation (allo-BMT). To analyze the expression of ICOS on
donor T cells, we transferred CFSE-labeled donor T cells into
irradiated allogeneic recipients and observed an increased expres-
sion of ICOS on alloreactive T cells compared to non-alloreactive
T cells. We then performed GVHD experiments in two models
with full MHC class I and II disparity and observed signiﬁcantly
less GVHD morbidity and mortality in recipients of ICOS
/

donor T cells. Interestingly, in GVHD/graft-versus-tumor (GVT)
experiments, ICOS
/
 donor T cells displayed intact GVT ac-
tivity, while their GVH activity was diminished. We are currently
performing experiments to assess the alloreactivity of ICOS
/
 T
cells in vivo, the inﬁltration of ICOS
/
 donor T cells into
GVHD target organs, and the degree of GVHD damage in target
organs by histopathological analysis. In conclusion, we have found
that ICOS
/
 donor T cells cause less GVHD morbidity and
mortality in comparison to wild-type donor T cells while preserv-
ing GVT activity. This data suggests that strategies to inhibit
ICOS could be useful for the prevention and/or treatment of
GVHD in recipients of an allo-BMT.
126
ADOPTIVE TRANSFER OF PRIMED T CELLS PRIOR TO ABLATIVE CON-
DITIONING AND BMT RESULTS IN SYNGENEIC ANTI-TUMOR IMMU-
NITY EARLY POST-TRANSPLANT
Jones, M., Jones, A.M., Levy, R.B. University of Miami School of
Medice, Miami, FL.
We have shown that mice vaccinated against a B6 myeloid leu-
kemia cell line (MMB3.19) prior to BMT followed by tumor
challenge post-BMT exhibited tumor speciﬁc immunity. Vacci-
nated BMT recipients exhibited prolonged survival compared to
unvaccinated BMT recipients indicating that anti-tumor reactive T
cells survived the conditioning regimen and subsequent autologous
BMT. To investigate T cell survival and function early after abla-
tive conditioning and BMT, an adoptive transfer BMT model
(AT-BMT) was developed. Since memory cells appear less suscep-
tible to irradiation induced apoptosis vs. naive cells, B6-gfp mice
were immunized with irradiated MMB3.19 cells 5-6 weeks before
the BMT as a source of anti-tumor primed T cells for adoptive
transfer (AT). Two days prior to BMT, 2.5x107 CD3 cells from
vaccinated donor B6-gfp mice were injected (i.p.) into naive
C57BL/6 (B6-gfp-) recipients. One day later (D-1), recipient mice
were conditioned with 8.5Gy TBI and at D 0 were injected with
TCD syngeneic B6-gfp- marrow. On D1 post-BMT, mice re-
ceived an ip challenge of 2.5x104 viable MMB3.19 cells. AT-BMT
recipients exhibited signiﬁcantly increased MST vs. non-ATBMT
controls (control: MST  13.8 days vs. AT-BMT: MST  40
days). To examine if so-called “low quality” CD8 T cells could
protect in this model, B6-wt and B6-CD4
/
 mice were primed 6
weeks prior to AT. Recipients received an AT of 2.5x107 CD4
and CD8 B6-wt (CD8  1.2x107) or CD4-CD8(1.2x107 cells)
on D-2 and were irradiated, transplanted and challenged with
tumor as above. In contrast to B6-wt T cells, recipients of CD4-
CD8T cells were not protected from tumor challenge. Following
transplant, spleen and peritoneal exudate cells (PEC) were ana-
lyzed for the presence of surviving AT cells. Transferred B6-gfp
cells were readily detected at Days4 and11 in the spleen (0.5%
and 1.5%, respectively) of AT-BMT recipients injected with tumor
post-BMT. Notably, a large percentage of gfp cells were identi-
ﬁed in the PEC of these AT-BMT mice at these time points
(10.7% and 50.9%, respectively). In total, these results demon-
strate that memory T cells can survive ablative BMT conditioning
and contribute to an effective immune response (i.e., anti-tumor) in
the early post-transplant period during lymphohematopoietic re-
constitution. These observations suggest that approaches to in-
crease selective host T cell survival during BMT may be useful to
diminish immune deﬁciency in the immediate post-transplant pe-
riod.
127
UNRELATED DONOR STEM CELL TRANSPLANTATION DOES NOT RE-
SULT IN LONG TERM REMISSION IN PATIENTS WITH MULTIPLE MY-
ELOMA DESPITE THE PRESENCE OF CHRONIC GVHD
Reynolds, C.M., Ratanatharathorn, V., Ayash, L.J., Silver, S.M., Daw-
son, L.A., Ferrara, J.L.M., Uberti, J.P. Univ of Michigan Stem Cell
Transplantation Program, Ann Arbor, MI.
Autologous SCT increases survival for patients with multiple
myeloma compared to conventional chemotherapy but is not con-
sidered curative as most patients eventually relapse. Allogeneic
SCT improves the CR rate and can result in a graft versus myeloma
(GVM) effect, but not all patients have a related donor. We
performed unrelated SCT using either a full or reduced intensity
preparative regimen. Patients with stage II or III multiple myeloma
who had relapsed following at least one prior therapy were eligible.
From January 1996 to May 2001 thirteen patients were enrolled.
Prior to SCT, patients had received a median number of 3 prior
regimens (range, 1-5). Six patients had previously received radia-
tion, and three had previously undergone an autologous transplant.
The median age at transplant was 50 years (range 38-62). Six
patients received a full intensity preparative regimen of total body
irradiation (TBI), busulfan, and cyclophosphamide, and seven pa-
tients received a reduced intensity regimen of total lymphoid irra-
diation, ﬂudarabine, and busulfan. All patients received HLA-
matched, unrelated donor BM (n  7) or peripheral stem cell (n 
6) transplants. The median mononuclear cell dose was 3.4 x 108 per
kg (range, 1.2–7.0). GVHD prophylaxis consisted of tacrolimus
and methotrexate. Three patients developed grade 3 aGVHD and
six developed cGVHD (1 limited, 5 extensive). The median fol-
lowup for all patients was 26.6 months (range 0.5–77.8). At the
time of SCT, nine patients had chemosensitive disease and two
patients were in CR. After transplant, six patients were in CR, ﬁve
had a PR and one had a stable monoclonal restriction. Seven
patients developed progressive disease at a median of 20.8 months
(range 3.2–39.8) post-transplant, four in the setting of extensive
cGVHD. Ten patients died (5 from relapse, 3 GVHD, 1 VOD,
and 1 pneumonia/ARDS). Three patients are alive with stable,
asymptomatic monoclonal restrictions at 26.6, 36.8, and 77.8
months after SCT. The Kaplan-Meier estimates at 1, 3 and 5 years
for overall survival were 54%, 26%, and 13 %, and for progression-
free survival were 54%, 15% and 7%, respectively. In this popu-
lation of heavily pretreated patients, unrelated donor transplanta-
tion did not result in long-term disease control despite the
presence of cGVHD and suggests that the GVM effect was not
sufﬁcient to prevent relapse.
128
UNRESTRICTED CTL RECOGNITION OF HFE, AN EMPTY-PEPTIDE–
GROOVE MHC CLASS I MOLECULE CONTROLLING IRON METABOLISM
HIGHLIGHTS ITS POTENTIAL ROLE AS A MAJOR HISTOCOMPATIBILITY
ANTIGEN IN UNRELATED BONE MARROW TRANPLANTATION
Rohrlich, P.S.1, Firat, H.1, Fazilleau, N.1, Ehrlich, R.2, Gluckman, E.3,
Loiseau, P.3, Lemonnier, F.1 1. Institut Pasteur, Paris, France; 2.
University, Tel-Aviv, Israel; 3. Hopital Saint Louis, Paris, France.
Unrestricted cytolytic T lymphocytes (CTL) responses against
human HFE, a non classical MHC class I molecule with an empty
peptide groove which controls iron metabolism, were induced in
HLA-A*0201 transgenic/H-2 class I knock-out mice. HFE recog-
nition was mediated by the alpha-beta T cell receptor. Similar
unrestricted CTL responses against mouse HFE molecule were
induced in DBA/2 HFE knock-out mice. Predominant usage of AV
6 segments was documented, contrasting with diversiﬁed V
segment usage. Thus absence of bound ligand to a histocompati-
bility class I molecule does not preclude  TCR interaction.
These results support the hypothesis that the immune system could
participate in the control of iron metabolism and indicate a novel
unrestricted allogenic recognition mechanism that could explain
the high proportion of allogenic cytotoxic precursors encountered
in any individual.
Ten to 15% of the individuals carry a mutated H41D HFE
molecule. Since HFE is encoded on the chromosome 6 and thus
cosegregates with the HLA haplotypes in siblings, a HFE mis-
Poster Session I
51BB&MT
match in related transplant is unlikely. In contrast, investigating
the frequency of a donor-recipient HFE mismatch in a series of 60
unrelated bone marrow transplants otherwise perfectly matched on
the HLA-A,B,C,DRB1,DQB1 loci, we found a mismatch in 10
cases. These data suggest that HFE could be a potent histocom-
patibility antigen in humans in view of its structural polymorphism.
129
MULTIDISCIPLINARY APPROACH TO DEVELOPMENT OF A GVHD OUT-
PATIENT CLINIC
Yount, P.1, Vance, E.A.1, Agura, E.D.1, Berryman, R.B.1, Fay, J.W.1,
Pineiro, L.1, Bengtson, E.1, Bair, J.D.2 1. Baylor Sammons Cancer
Center and Texas Oncology, P.A., Dallas, TX; 2. Baylor University
Medical Center, Dallas, TX.
Managing GVHD is a particularly challenging aspect of the care
of blood and marrow transplant recipients. In the Blood and
Marrow Transplant program at Baylor University Medical Center,
the GVHD patients were identiﬁed as a population that could
beneﬁt from an interdisciplinary approach in a specialized clinic
setting. A questionnaire was sent to the transplant physician group
to gauge interest in and the need for this clinic. It was determined
that enough interest was present and a patient population would
support the organization of this program. The interdisciplinary
team consisted of an oncologist as team leader, dermatologist,
clinic manager, contract administrator, nurse manager, pharmacist,
biostatistician, physical therapist, occupational therapist, dietitian,
enterostomal therapist, social worker, and chaplain. The team
gathered for regular meetings to decide on structure, services, and
supplies that would be needed to support such an endeavor. The
clinic manager and contract administrator investigated ﬁnancial
reimbursement for each discipline in the outpatient setting. Orga-
nization of the GVHD clinic recommended that each patient see
the medical team leader followed by other specialty areas that
would beneﬁt the patient. A key component for a successful clinic
is data collection for patients before their appointments to assist
clinicians with a thorough needs assessment. Each discipline of-
fered a data collection tool and these were compiled by a team
member. This tool includes sections on medication, medical symp-
toms, skin assessment, nutrition, physical and occupational ther-
apy, and psychosocial needs. Prior to patient’s appointments, ques-
tionnaires are mailed with the instructions to complete and bring
to their appointment. Research protocols for disciplines in this
clinic are being developed. Taking a multidisciplinary approach to
a GVHD clinic is a key factor in developing a successful program.
130
IL-7 IS NECESSARY FOR THE DEVELOPMENT OF EXPERIMENTAL
GRAFT-VERSUS-HOST DISEASE (GVHD)
Chung, B., Dudl, E.P., Toyama, A., Barsky, L., Price, M., Weinberg,
K.I. Childrens Hospital Los Angeles, Los Angeles, CA.
Interleukin 7 (IL-7) promotes both thymopoiesis and the survival
and proliferation of mature T lymphocytes. Although IL-7 en-
hances immune reconstitution following hematopoietic stem cell
transplantation (HSCT), its effect on mature T lymphocytes in
allogeneic bone marrow transplantation (BMT) could lead to ex-
acerbation of GVHD. However, recent experiments to determine
whether IL-7 treatment worsens GVHD have produced conﬂict-
ing results. Therefore, this study was designed to examine whether
IL-7 is necessary for the induction of GVHD by maintaining
donor mature T cells following allogeneic BMT. In order to
induce GVHD, B6 or B6/IL-7
/
 (CD45.2 H2Kb) recipient
mice were lethally irradiated (1300 cGy) and co-transplanted with
4x106 lymph node (LN) and 1x106 T cell depleted (TCD) BM cells
from either congenic B6.SJL (CD45.1, H2Kb) or major MHC
mismatched allogeneic Balb/C (CD45.2, H2Kd) donor mice. Fol-
lowing transplantation, the recipient mice received either human
recombinant IL-7 (rhIL-7) 500ng BID or PBS from day 1 to 60.
The survival rate was similar in all groups of mice for the ﬁrst 25
days after transplantation and no evidence of GVHD was detected
from allogeneically transplanted B6/IL-7
/
 recipients treated
with PBS. In contrast, GVHD-related mortality and morbidity in
allogeneic recipients treated with IL-7 were increased compared to
the PBS treated groups. IL-7 treatment signiﬁcantly reduced sur-
vival in the B6/IL-7
/
 mice (15%, n  20) compared to the PBS
treated B6/IL-7
/
 recipients (59%, n 17 [p 0.005]). Further-
more, the overall GVHD clinical index of IL-7 treated B6/IL-7
/

was signiﬁcantly lower than the PBS treated B6/IL-7
/
 recipients
(p  0.05). The recovery of donor CD4 or CD8 T cells in the
periphery of the PBS treated B6/IL-7
/
 recipients was signiﬁ-
cantly lower than the IL-7 treated B6/IL-7
/
 mice by day 30
post-transplantation. In addition, IL-7 treatment increased the
number of activated (CD69 positive) donor-derived CFSE labeled
CD4 and CD8 LN T lymphocytes in the lymph nodes of B6/IL-
7
/
 mice compared to PBS treated B6/IL-7
/
 recipients indi-
cating that IL-7 enhances maintenance of dividing activated al-
logneic mature T cells. Therefore, it is likely that IL-7 is necessary
for the development of GVHD presumably by maintaining the
adequate number of activated donor T cells in the periphery of the
recipient animals post-allogeneic BMT.
131
THE ABSENCE OF PEYER’S PATCHES OR HOST TNF- DOES NOT
AFFECT GVHD PROGRESSION IN AN ALLOGENEIC BMT MODEL USING
EXTENSIVE CONDITIONING REGIMENS
Welniak, L.A.1, Kuprash, D.V.2, Tumanov, A.V.2, Sun, K.1, Stull,
S.W.2, Anver, M.R.2, Nedospasov, S.A.2, Murphy, W.J.1 1. University
of Nevada, Reno, Reno, NV; 2. SAIC-Frederick, Frederick, MD.
Peyer’s patches (PP) have been previously shown to play a critical
role in the initiation of acute lethal GVHD in a non-irradiated
murine model (Murai et al., 2003). This report utilized IL-7
receptor antibody treatment of pregnant mice to inhibit the devel-
opment of PPs in the fetus. We wanted to assess GVHD progres-
sion in mice congenitally deﬁcient in PP presence and function.
The absence of various members of the TNF and TNF-R families
also affects secondary lymphoid structure and organization. The
presence of PP has varied between different independently gener-
ated TNF knockout strains which may be related to the close
linkages of tnf with lta and ltb genes. A new TNF deﬁcient strain
has been developed that lacks PPs, displays the defects character-
istic of TNF ablation but not the LT associated defects character-
ized by lack of lymph nodes and defects in splenic microarchitec-
ture. To examine the role of host-derived TNF family members in
acute lethal GVHD, we also had to assess the role of secondary
lymphoid structures. We utilized a full MHC mismatched mode of
BALB/c (H2d) cells into myeloablated C57BL/6 (H2b) recipients.
We lethally irradiated wild type (WT) mice, TNF- deﬁcient mice
with Peyer’s patches (PP) and TNF- deﬁcient mice without
Peyer’s patches (PP-) followed by infusion of allogeneic bone
marrow and spleen cells. We observed no difference in the survival
of the three groups of mice (median day of death ranged from 26
to 27). In addition, no signiﬁcant differences were observed in
GVHD associated histopathological lesions in the small intestine.
Hisopathological lesions observed in the colon were similar in both
the PP and PP- TNF- deﬁcient recipients. Based on our ob-
servations, we conclude that (1) Peyer’s patches and (2) host
TNF- are not required for the development of acute lethal
GVHD in mice that received extensive conditioning and allogeneic
bone marrow transplantation.
132
DEVELOPMENT OF NUTRITION ALGORITHMS FOR CHRONIC GVHD
Bair, J.D.1, Roberts, S.R.1, Thompson, J.1, Agura, E.D.2, Berryman,
R.B.2, Fay, J.W.2, Pineiro, L.2, Vance, E.A.2 1. Baylor University
Medical Center, Dallas, TX; 2. Baylor Sammons Cancer Center and
Texas Oncology, P.A., Dallas, TX.
One of the challenges for patients with chronic GVHD is to
maximize quality of life by maintaining optimal nutritional intake
and minimizing loss of lean body mass. Registered Dietitians
(RD’s) play an integral role in the nutritional care of patients in the
Blood and Marrow Transplant program at Baylor University Med-
ical Center and are part of the team establishing a new GVHD
clinic. Patients referred to the GVHD clinic are seen by the RD
Poster Session I
52


Unrestricted CTL recognition of HFE, an empty-peptide-groove MHC class I molecule controlling iron metabolism highlights its potential role as a major histocompatibility antigen in unrelated bone marrow tranplantation 

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Unrestricted CTL recognition of HFE, an empty-peptide-groove MHC class I molecule controlling iron metabolism highlights its potential role as a major histocompatibility antigen in unrelated bone marrow tranplantation

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