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`VOLUME 114
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`ed =) a a4
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`20 NOVEMBER 2009
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`Fifty-first annual meeting
`abstracts
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`December 5-8, 2009
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`New Orleans, Louisiana
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`Miltenyi Ex. 1012 Page’2°
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`

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`
`
`COVERFIGURE
`
`20 NOVEMBER 2009 - VOLUME 114, NUMBER 22
`
`CONTENTS
`
`Jazz musician with trumpet
`
`
`
`
` blood
`
`INDEX TO
`ABSTRACTS
`
`xiii
`
`Index to Abstracts
`
`JOURNAL OF
`
`THE AMERICAN
`SOCIETY OF
`
`HEMATOLOGY
`
`ABSTRACT
`REVIEWERS
`
`
`
`xxiii
`
`Reviewers
`
`xxv Abstract Policy and Abstract Description
`
`xxvi
`
`Blood Editorial Board
`
`ABSTRACTS
`
`1
`
`7
`
`Plenary Session
`
`OralSessions
`
`405
`
`Poster Sessions
`
`1573
`
`Invited Scientific Program
`
`AUTHOR INDEX
`
`1589 Author Index
`
`KEYWORD/SUBJECT
`INDEX
`
`
`
`1679 Keyword/Subject Index
`
`KEY
`
`VV This icon denotes an abstract that is clinically relevant.
`
`Small caps denote presenting author.
`Asterisk [*] with author names denotes non-ASH members.
`
`
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`Miltenyi Ex. 1012 Page 3
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`Miltenyi Ex. 1012 Page 3
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`LYMPHOMA: THERAPY WITH BIOLOGIC AGENTS, EXCLUDING PRE-CLINICAL MODELS
`Abstract 929
`
`383
`
`linked to an IgG4 Fe domain,the T cell receptor CD3 zeta chain and a CD28 costimulatory
`domain. The specificity and function of the ROR1 CAR was comparedwith a similarly
`Adoptive Cell Therapy for Lymphoma:Use of CpG-Loaded TumorCells to
`designed CARspecific for the CD20 molecule, which is expressed on both malignant and
`Generate Potent Anti-Tumor CD4 T Cell Immunity. MatrHew J GoLp-
`normalBcells, and is being targeted with gene-modified T cells in clinical trials. Primary
`STEIN*, Bindu Varghese, PhD*, Ranjani Rajapaksa, PhD*, Joshua Brody, MD,
`human CD8* T cells were transduced with the ROR1 and CD20-specific CARs Trespec-
`Shoshana Levy, PhD* and Ronald Levy, MD, Division of Oncology, Stanford
`tively, and T cells expressing high levelsof the receptors were sort-purified using an anti-Fe
`School of Medicine, Stanford, CA, USA
`antibody. T cells that expressed either the ROR1-specific CAR or the CD20-specific CAR
`Background: Recently, we have investigated adoptive cell therapy for treating lym-
`efficiently lysed primary B-CLL samples (5/5) obtained from patients with advanced
`phoma. Theefficacy of this maneuver has been demonstrated by curing large established
`disease, and also lysed a MCLcell line (JeKo-1), and a ROR1+ B-ALLcell line (BALL-1).
`tumors. Specifically, we use active immunization to generate anti-tumorT cells in vivo and
`ROR1-specific T cells did not recognize the myeloid leukemiacell line K562,but efficiently
`transfer
`these T cells into lymphodepleted recipient mice (Brody J, Goldstein MJ,
`lysed K562 cells that had been transfected to express RORI, confirming the specific
`Czerwinski DK, and Levy R; Blood, 2009). A major challenge in adoptive therapy is the
`recognition of RORI ontargetcells. Consistent with the expression pattern of the target
`method of generating anti-tumorT cells. Traditionally, tumor-specific T cells are expanded
`molecules, T cells that expressed the CD20-specific CARalso efficiently lysed normal
`to large numbers ex vivo. Herein, we describe a new, whole-cell vaccinethatis effective in
`primary and EBV-transformedB cells, but T cells that expressed the ROR1-specific CAR
`inducing anti-tumorT cells in vivo. This vaccine combines tumorantigens with an immune
`did not recognize nonmalignant or EBV-transformedB cells. Activation of normalB cells
`stimulant: irradiated-tumor cells (a source of tumor antigens) are loaded with the TLR
`by engagement of the B cell receptor or activation through CD40 induced B cell
`agonist CpG (an immunestimulant). Our vaccine approach has several potential advan-
`proliferation and upregulation of the CD80 and CD86 costimulatory molecules, but did not
`tages:
`(1) anti-tumor immunity generated by our CpG-loaded, whole-cell vaccine is
`result in RORI surface expression by flow cytometry or recognition by T cells that
`poly-antigenic and thus, not limited by the expression ofa single antigen target on tumor
`expressed the RORI-specific CAR. These results suggest
`that
`targeting RORI with
`cells; (2) ex vivo expansion may generate large numbersofeffector T cells that can induce
`gene-modified T cells may have advantages over targeting B cell-lineage restricted
`tumor regression in the short-term, but have a limited ability to maintain a persistent
`molecules such as CD19 and CD20that are expressed on normal mature B cells. Studies to
`anti-tumor response. Our model avoids ex vivo manipulation of anti-tumorT cells and thus,
`determine whether ROR] is expressed during a stage of normal B cell developmentare in
`maypreserve and enhance a memoryTcell population thatsustains the anti-tumorresponse.
`progress. ROR] is highly conserved in non-humanprimates and this model may be suitable
`Methods: We derived a new pre-B cell lymphomacell line in the CS7BL/6 background.
`to determine potential toxicities of adoptive immunotherapy with ROR1-specific CAR
`Primary bone marrowcells were isolated from C57BL/6 donor mice andtransfected with a
`expressing T cells.
`recombinantretrovirus containing the Ber-Abl oncogene. The emerging transformed cell
`Disclosures: Norelevantconflicts of interest to declare.
`line was designated H11. This cell line expressed the B lineage marker CD19 but was
`negative for MHCII and surface Ig. Irradiated H11 tumorcells were pre-loaded with CpG
`for 24 hours and administered to donor mice by daily, sub-cutaneousinjections for five
`days. Donor splenocytes were harvested seven days following vaccination and adoptively
`transferred into lethally irradiated recipient mice that were subsequently challenged with a
`lethal dose of H11 tumorcells,
`Results: Vaccination with CpG-loaded H11 tumor cells (CpG-H11) generated anti-
`tumor T cells that are effective in adoptive cell therapy. 100% of mice receiving adoptive
`therapy with vaccine-induced T cells were protected from tumorchallenge. In contrast,
`“Hiper CVAD Followed by Rituximab Purging Previous to Autologous
`vaccination of donor mice with untreated H11 tumor was insufficient for generating
`Stem-Cell Transplantation as Therapy of Mantle Cell Lymphoma.Results
`anti-tumorT cells. Only 20% of mice treated with T cells from these donors were protected
`of Manto 2000 study”. F.J. Capote*!, Eva Gonzalez-Barca*?, Albert Oriol,
`from tumor challenge. In spite of the H11 tumor being MHCClassII-, we observedthat
`anti-tumor immunity generated by the CpG-H11 vaccine was CD4Tcell mediated. CD4 T
`MD*3, Antonio Romero-Aguilar*4, Angel Leon*5, F.J. Fernandez-Calvo*®,
`cells were isolated from CpG-H11 vaccinated donors by flow cytometry. Fewer than
`Elena Pérez-Ceballos*”, Eva Donato*®, Dolores Caballero*®, Jose Antonio
`1.8X 10° CD4T cells were sufficient to protect 80% ofrecipient mice from tumorchallenge.
`Queizdn*!°, Marfa Jesus Pefiarrubia, MD*!!, Luis Palomera*!2,Julio Prieto*!3,
`In contrast, equivalent numbers of donor CD8Tcells provided no benefit. These results
`Pilar Giraldo, MD" and JuaN BERGuA, MD*)3, !Hospital Puerta del Mar.
`strongly suggest that the CpG-H11 vaccine inducedcross-presentation of tumorantigens by
`Cadiz, Cadiz, "Hematology, Hospital Universitari de Bellvitge, L’Hospitalet de
`antigen-presenting cells (APCs). We have demonstrated that CpG-loaded H11 tumorcells
`Llobregat, Spain, *Hospital Universitari Germans Trias i Pujol, Badalona,
`can OleakO CpG into the immediate environmentactivating nearby APCs. These APCs
`Spain, ‘Hematology, Hospital Virgen de las Nieves. Grandad, Granada, Spain,
`have greater phagocytic potential and express higher levels of co-stimulatory molecules
`SHospital Univ. La Fe. Servicio de Hematologia, PETHEMA cooperative group,
`such as CD40. Ongoing studies will determine whether APCs which encounter the
`Valencia, Spain, °Hematology, Clinico. Valladolid, Vallodolid, Spain, 73M
`CpG-H11 vaccine but not untreated H11 tumorcells, can stimulate proliferation of
`anti-tumorT cells.
`Failure Spanish Study Group (Pethema-GETH), Murcia, Spain, *Hospital
`General de Castellén, Castellén, °Hematology-BMT Unit, Hospital Clinico
`Salamanca, Salamanca,Spain, '°Hospital General de Segovia, Segovia, !\HOS-
`PITAL RIO HORTEGA,Valladolid, Spain, '*Hospital Lozano Blesa, Zaragoza,
`Spain, Hematology. Hospital San Pedro de Alcdntara. Caceres, PETHEMA
`cooperative group, Caceres, Spain, '*Hospital Miguel Servet, Zaragoza, Spain
`Purpose: Multicentre phaseII study in newly diagnosed Mantle cell lymphomapatients
`to determine the feasibility, overall response (OR) and failure free survival (FFS) of
`intensive chemotherapy type Hyper-CVADfollowed by in vivo purging with Rituximab
`CD8* T Cells Engineered to Express a ROR1-Specific Chimeric Antigen
`previousperipheral stem-cell transplantation.
`Receptor Specifically Recognize ROR1 Positive B Cell Tumors. MICHAEL
`Treatment scheme consisted in four cycles of Hyper-CVAD chemotherapy as is
`HubEceK, MD*!, Thomas M Schmitt, PhD*!, Sivasubramanian Baskar, PhD*2,
`described by Romaguera et al. After chemotherapy four weekly Rituximab courses
`(375mg/m2) were administrated previously to peripheral stem-cell collection. Rituximab
`Wen-Chung Chang*?, David G Maloney, MD, PhD!, Michael C Jensen, M.D.*3,
`was not addedatthe sametime as chemotherapy cycles; its role consisted in working as a
`Christoph M. Rader, Ph.D.*? and Stanley Riddell, MD*!,
`'Clinical Research
`purging agent previous stem-cell mobilization. After Rituximab administration peripheral
`Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA,
`blood progenitors were collected. Mobilization was performed using Cyclophosphamide
`Experimental Transplantation and Immunology Branch, Center for Cancer
`plus G-CSFat dose of 10 g/Kg/day.If the first mobilization was unsuccessful, a second
`Research, National Cancer Institute, NIH, Bethesda, MD, USA, 3Division of
`schemewas used, using Ifosfamide (10 g/m2 in 72 hours infusion on day 1), Mesna(10
`Cancer Immunotherapeutics & Tumor Immunology, City of Hope National
`g/m2 days 1 and 2), VP16 (150 mg/m2days 1 to 3) plus G-CSFat doseof 5 pg/Kg each 12
`Cancer Center, Duarte, CA, USA
`hours. The aim was obtain 2x 106 CD34 cells/Kg. After mobilization peripheral autologous
`The orphan tyrosine kinase receptor ROR1 was previously identified as a highly
`stem-cell
`transplantation (PASCT) was performed using BEAM (BCNU,Ethoposide,
`expressed gene by expression profiling of B cell chronic lymphocytic leukemia (B-CLL),
`Ara-C and Melphalan)as conditioning regimen..A weekafterplatelet recovery (>50109/L)
`[Klein et al. J Exp Med 2001], and has subsequently been shownto be expressed on mantle
`another four weekly Rituximab courses (375mg/m2) were added. Patients were followed
`after treatmentin each centre.
`cell lymphoma (MCL)anda subset of B cell acute lymphoblastic leukemias (B-ALL).
`ROR1 encodes a 105 kDa protein that contains Ig-like, cysteine rich, kringle, tyrosine
`Forty-four patients diagnosed of mantle cell lymphomaandpreviously nottreated were
`kinase and proline rich domains and is expressed during embryonic development butis
`enrrolled from fifteen Spanish Institutions from 2000 to 2006. The median age of the
`absent on normal adult tissues including non-malignant B cells. The function of RORI in
`patients were 55.77 year old. Male/female rate was 3:1. Forty patients had an Ann-Arbor
`normal and malignant cells is not known, although secreted Wnt proteins have been
`stage IV, and gastrointestinal
`involvement was present
`in twenty-nine. Marrow was
`proposed as candidate ligands. Analysis of ROR protein expression using specific
`infiltrated in 83.3% of the cases. Age IPI adjusted were =2 in 45% ofthe cases respectively
`polyclonal antibodies revealed uniform, stable, and restricted cell surface expression on
`(table 2). Blastic mantle cell lymphoma was diagnosedin 5 patients (11.9%). The median
`B-CLL, suggesting this molecule is a candidate for targeted immunotherapy ofBcell
`follow-up of the patients was 75,07 months. In the intention to treat, of the forty-four
`malignancies[Baskaret al. Clin Cancer Res 2008]. We constructedalentiviral vector that
`patients only 26 patients receive all the treatment (59%). Autologous peripheral stem-cell
`encodes a chimeric antigen receptor (CAR) consisting of single chain variable (scFV)
`transplantation was notperformedin 16 patients. The causes of not complete the treatment
`fragmentsof the heavy andlight chains of a murine monoclonal antibody specific for ROR1,
`schedule was: three patients refuse to be transplanted; mobilization failure in four; death
`
`
`
`
`
`
`LYMPHOMA: THERAPY WITH BIOLOGIC AGENTS, EXCLUDING
`PRE-CLINICAL MODELS: TREATMENT OF B-CELL LYMPHOMASWITH
`
`Anti-CD20 ANTIBODIES
`
`Abstract 931
`
`induces
`Conclusions: Here we describe a novel, whole-cell vaccine approach that
`anti-tumor T cells for adoptive therapy to treat lymphoma. This vaccine is superior to
`vaccination with tumorcells alone. Weare currently developingthis therapy for evaluation
`in a clinical trial to treat mantle cell lymphoma.
`Disclosures: No relevant conflicts of interest to declare.
`
`Abstract 930
`
`2009 ASH ANNUAL MEETING ABSTRACTS, VOLUME 114, ISSUE 22, NOVEMBER20, 2009
`
`Miltenyi Ex. 1012 Page 4
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