Andre, M.C.

Handgretinger, R.

Hartwig, U.F.

Hemmerling, J.

Herr, W.

Hoffmann, J.

Huber, C.

Meyer, R.G.

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130 Poster Session-IISR-GVHD, defined as lack of response or disease progression after
at least 7 days of treatment with methylprednisolone at 2 mg/kg. Be-
tween 3/2003 and 6/2008, 20 patients with advanced SR-GVHD fol-
lowing allogeneic transplantation were treated with DD at OHSU, 7
afterMRD and 13 after URD (six matched, one A antigenmismatch,
and six allele mismatch) .GVHD prophylaxis consisted of cyclo-
psporine, methotrexate, and steroid (19) or cyclosporine plus
MMF (1). Nineteen patients developed acute SR-GVHD overall
grade III (7) and IV (12) while one patient developed chronic exten-
sive SR-GVHD. Twelve patients received DD as first salvage ther-
apy while 8 patients were treated with other agents prior to DD (6
had one and 2 had two prior salvage therapies including MMF and
Etanercept). Planned treatment was DD at a dose of 600 mcg
(9mcg/kg) on days 1, 3, 5, 15, 17, and 19.Median days toDD therapy
after stem cell transplant was 38.Ten patients showed response (4CR
and 6 PR defined as at least a one overall grade improvement), six
were non responders, and four patients died before completion of
treatment. One patient remains alive. Causes of death included
GVHD (8), opportunistic infection (5), respiratory failure (2), trans-
plant-associated microangiopathy (2), ICH (1), and relapse (1). Me-
dian survival for the entire group was 30.5 days from the onset of DD
(4–18391) with median survival of 116 days for patients who re-
ceived all 6 doses of DD. Despite measurable responses in 50% of
patients, no survival benefit was observed. Profound immune sup-
pression was significant among responders, contributing to opportu-
nistic infection and death. Thus, DD may be most beneficial if used
earlier in the course or in lower grade aGVHD. Responses can be
achieved in advanced SR-GVHDwithDDbut novel approaches still
remain the research target for these patients given poor overall
survival .362
EVALUATION OF ALLOIMMUNE RESPONSES IN HUMANIZED NOD/SCID/
IL2RGNULL MICE FOLLOWING HUMAN CD341 STEM CELL TRANSPLAN-
TATION
Hartwig, U.F.1, Hemmerling, J.1, Hoffmann, J.1, Andre, M.C.2,
Handgretinger, R.2, Huber, C.1, Herr, W.1, Meyer, R.G.1 1Johannes
Gutenberg-University School of Medicine,Mainz, Germany; 2Eberhard--
Karls University School of Medicine, Tuebingen, Germany
Immunodeficient NOD/SCID/IL2Rg (yc)null mice can be effi-
ciently used to establish human lymphohematopoiesis or lymphohe-
matological neoplasia in a murine environment. Thus, upon
engraftment of human CD341 hematopoietic stem cells (HSC) or
acute leukemia cells and allogeneic donor lymphocyte infusions
these mice might become a powerful tool to provide an allogeneic
HSCT model for investigating human graft-vs-host (GvH)- and
graft-vs-leukemia (GvL)-immunity in vivo. In the current study,
we engrafted NOD/SCID/ycnull mice with human CD341 HSC to
provide a model for analyzing the role of patient-derived antigen
presenting cells for the induction of GvH- and GvL-reactivity and
to evaluate reactivity to human alloantigens of ex vivo engineered
T cell allografts for improved immunotherapy. Following reconsti-
tution of busulfan conditioned neonatal or (300 cGy) irradiated adult
NOD/SCID/ycnull recipients with 1  105 to 1  106 purified
CD341 adult HSC and weekly i.v. injections of Fc-IL7, human
CD451 PBMNC were detected as early as early as 30 or 60 days
(d) post reconstitution of newborn and adult mice, respectively. After
120d, T-, B-, and NK-cells, as well as monocytes and dendritic cells
(DCs) were found in spleen (SPL) and bonemarrow of both newborn
and adult recipients by flow cytometry and immunhistological stud-
ies in e.g. SPL and skin. However, as reconstitution of newborns re-
sulted in thymopoiesis of human CD31 cells and better lympho- and
myelopoiesis as compared to adult mice, further studies were per-
formed in HSC engrafted newborn recipients. Functional analyses
of lymphocytes grown in humanized NOD/SCID/ycnull mice re-
vealed that human CD31 T cells isolated from SPL and challenged
ex vivo with i) murineNOD/SCID/ycnull derivedDCs, ii) HLA-mis-
matched DCs or iii) HSC donor derived (autologous) DCs elicited
alloresponses to human but not murine alloantigens(s). These results
and further studies on ex vivo examined allo- and xenoimmunity of
naı¨ve HLA-matched and HLA-mismatched CD81 T cells trans-
ferred into HSC engrafted NOD/SCID/ycnull recipients will be pre-
sented to demonstrate T cell alloimmune responses in humanized
mice and to show that our model can be used to evaluate residualGvH reactivity of CD81 donor T cells in HLA- matched or haploi-
dentical settings. Finally, these studies should help to elucidate the
impact of xenoreactivity on the induction of GvH immunity by hu-
man T cells in humanized mice.363
PROPHYLACTIC ADMINISTRATION OF EX VIVO CO-STIMULATED DONOR
LYMPHOCYTE INFUSION (DLI) FROM RELATED AND UNRELATED
DONORS AFTER REDUCED INTENSITY CONDITIONING FOR HIGH RISK
HEMATOLOGIC MALIGNANCIES
Goldstein, S.C., Levine, B., Smith, J., Hinkle, J., Luger, S., Perl, A.,
Loren, A., Frey, N., Stadtmauer, E., Porter, D.L., June, C.H. University
of Pennsylvania, Philadelphia, PA
Enhancing the graft versus tumor (GvT) effect without graft ver-
sus host disease (GvHD) is critical to the success of reduced intensity
(RIC) strategies for allogeneic transplantation. Despite establishing
donor lymphohematopoiesis, relapse rates remain high. Standard
DLI given at relapse or prophylactically to enhance donor chimerism
has met with limited success in improving transplant outcomes. Co-
stimulation of donor T-cells, using anti-CD3/CD28 coated beads to
serve as artificial APCs, may reverse functional T cell tolerance,
thereby restoring immune responsiveness and potentiate GVT. Un-
like standardDLI, ex vivo co-stimulationmay enhanceT cell activity
by removing and activating T cells outside of a tumor-induced im-
munosuppressive milieu. We report the preliminary results of a fea-
sibility trial of ex vivo co-stimulated DLI from sibling and unrelated
donors given ‘prophylactically’ at 4 and 6 months after RIC alloge-
neic transplantation for patients (pts) with high-risk hematologic
malignancy.
Methods: 15 pts have undergone RICwith alemtuzumab, Fludar-
abine,Busulfan with peripheral blood stem cell transplantation
(PBSCT) and planned activated DLI (pADLI) at 4 months (1 
107 CD31/kg) and 6 months (1  108 CD31/kg) post-transplant
in the absence of GvHD or relapse. All pts received tacrolimus and
methotrexate as GvHD prophylaxis.
Results: 8 pts (AML, n5 6; ALLCR1 n5 2) received grafts from
HLA-identical sibling donors, 7 (AML n5 5;MDS n5 1; ALL, n5
1) from matched unrelated donors. Donor hematopoiesis was estab-
lished in all patients. Of the 11 infusions given to date, there has been
no infusion-related toxicity, confirming the safety and feasibility of
this strategy. 8 pts have received the first of 2 planned infusions of
pADLI. Of the remaining 7 pts, 4 infusions are upcoming, 1 was pre-
cluded by early relapse, and 2 were precluded by early NRRM. Of
the 8 pts who received their initial pADLI#1, 3 have received a second
infusion. 5 pts did not receive pADLI#2 because of relapse (n 5 3),
GvHD (n5 1), and transient uveitis in the absence ofGvHD (n5 1).
Overall incidence of acute GvHD (aGvHD) has been low; 1 pt devel-
oped aGvHD prior to pADLI#1, and 1 pt developed aGvHD after
pADLI #1. No patient has developed chronic GvHD.
Conclusion: Preliminary results of this trial demonstrates that
RIC with PBSCT followed by ex vivo costimulated pADLI for
poor prognosis hematologic malignancies is safe and feasible with
potential for enhancing GvT without increasing GvHD.364
CLINICAL EXPANSION OF CORD BLOOD DERIVED T CELLS FOR USE AS
DONOR LYMPHOCYTE INFUSION AFTER CORD BLOOD TRANSPLANTA-
TION
Okas, M.1,2, Gertow, J.1,2, Uzunel, M.1, Karlsson, H.1, Uzunel, M.1,
Ringden, O.1,2, Mattsson, J.1,2, Uhlin, M.1,2 1Karolinska Institutet,
Stockholm, Sweden; 2Karolinska University Hospital, Stockholm, Sweden
Background: When no HLA-identical donor is available, cord
blood transplantation (CBT) is an attractive option due to the rapid
availability of the graft and its biological properties. One of the dis-
advantages associated with CBT is lack of possibility for donor lym-
phocyte infusion (DLI) after CBT. Here we report expansion and
characterization of CD3 positive lymphocytes from CB grafts in
connection to CBT.
Material andMethods:Lymphocytes from 13 CB grafts were ex-
panded with CD3/CD28 beads and 200 IU/ml rIL2. Expanded cells
were cryopreserved in ready-to-use DLI doses. Immunophenotyp-
ing and cytokine production assay were performed on expanded


Evaluation Of Alloimmune Responses In Humanized NOD/SCID/IL2rgnull Mice Following Human CD34+ Stem Cell Transplantation 

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Evaluation Of Alloimmune Responses In Humanized NOD/SCID/IL2rgnull Mice Following Human CD34+ Stem Cell Transplantation

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