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A PHASE 1 Perspective: Multivirus-Specific T CELLS from BOTH Cord Blood and BONE Marrow Transplant Donors
S140
Abstracts / Biol Blood Marrow Transplant 22 (2016) S19eS481
Health, Bethesda, MD; 7 Experimental Transplantation and Immunology Branch, National Cancer Institute, NIH, Bethesda, MD; 8 Medical Oncology, Greenebaum Cancer Center, Baltimore, MD; 9 Georgia Regents University Cancer Center, Augusta, GA; 10 Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD; 11 Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA; 12 Translational Research Programs, Center for Cellular Immunotherapies, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA; 13 Experimental Transplantation and Immunology Branch, National Institutes of Health, National Cancer Institute, Bethesda, MD We previously demonstrated that pre-emptive DLI after low intensity HCT using rapamycin-resistant donor CD4+ T cells (12-day manufacturing; T-Rapa12) led to successful engraftment, GVL effects, and a moderate rate and severity of GVHD (NCT0074490; Fowler et al, Blood, 2013); subsequent patients received T-Rapa cells manufactured in 6 days (T-Rapa6). All patients received an HLA-matched sibling mobilized allograft after conditioning with fludarabine (total dose, 120 mg/m2) and cyclophosphamide (total dose, 1200 mg/m2). GVHD prophylaxis consisted of cyclosporine plus short-course sirolimus (through d14 post-HCT). Donor T-Rapa cells (2.5 x 107/kg) were given on d14 post-HCT. For the current analysis, chemotherapy responsive patients with lymphoid malignancies (n¼62) that received transplant and either T-Rapa12 or T-Rapa6 cell DLI were pooled for evaluation. Recipients were stratified into prognostic groups I/II (low-intermediate risk) versus III (high risk) based on histology and remission status as shown in Table 1 (after Kahl et al, Blood, 2007). In our cohorts, no significant differences were observed between Kahl I/II vs. Kahl III risk groups in regards to GVHD, chimerism, or overall survival (OS; Table 1). Estimated OS at 36-months post-HCT in Kahl I/II and Kahl III T-Rapa cohorts was 74.6% and 67.6%, respectively (p¼0.32 overall) (Fig. 1a). By comparison, in the Kahl et al study, which differed from our analysis in part by the inclusion of unrelated donors, estimated OS at 36-months was clearly divergent for Kahl I/II vs. Kahl III diagnoses (I: 60% II:55% III:26%; Fig. 1b). Our data suggest that the addition of a preemptive T-Rapa DLI may improve on previously reported overall survival results in low intensity HCT for high-risk lymphoid malignancies such as DLBCL (not in CR) and HD.
Table I
Number of Patients Age: Median (Range) Diagnoses
Acute GVHD all Grades (%) Acute GAVHD Grade 3/4 (%) Late Acute GVHD (%) Chronic GVHD (%) Day 100 Full* T Cell Chimerism (%) Day 100 Full* Myeloid Chimerism (%) 36 Month OS (%) *Donor Chimerism 96%
Kahl I/II
Kahl III
p value
24 59.5 (35-68) Follicular (9) Mantle Cell (11) DLBCL CR (2) CLL noCR (2)
N/A <0.0001 N/A
33.3 4.2 17.4 56.5 37.5
38 47 (22-66) DLBL noCR (22) Hodgkin’s (11) Gray Zone (3) PTCL noCR (3) 34.2 7.9 24.8 40.0 50.0
70.8
73.0
1.00 NS
74.6
67.6
0.32 NS
0.90 1.00 0.53 0.28 0.60
NS NS NS NS NS
Figure 1.
168 A PHASE 1 Perspective: Multivirus-Specific T CELLS from BOTH Cord Blood and BONE Marrow Transplant Donors Patrick J. Hanley 1, Michael Daniel Keller 2, Maria Martin Manso 3, Caridad Martinez 4, Kathryn Leung 4, Conrad Russell Y. Cruz 5, Cecilia Barese 3, Sarah McCormack 6, Min Luo 3, Robert A. Krance 4, David A. Jacobsohn 7, Cliona M. Rooney 8, Helen E. Heslop 8, Elizabeth J. Shpall 9, Catherine M. Bollard 2. 1 CETI, Blood and Marrow Transplantation, Children’s National Medical Center, Washington, DC; 2 Center for Cancer and Immunology Research, Children’s National Medical Center, Washington, DC; 3 CETI, CNMC, Washington, DC; 4 Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children’s Hospital Cancer Center, Methodist Hospital Houston, Houston, TX; 5 Children’s National Health System, Washington, DC; 6 Children’s National Medical Center, Washington, DC; 7 Division of Blood and Marrow Transplantation Center for Cancer and Blood Disorders, Children’s National Health System, Washington, DC; 8 Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children’s Hospital, Houston Methodist Hospital, Houston, TX; 9 Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX CMV, EBV and adenovirus are problematic in patients after stem cell (SCT) and cord blood transplantation (CBT) and are associated with morbidity and mortality. Deficiencies in conventional therapeutics have increased interest in an immunotherapeutic approach to viral disorders. We have developed 2 strategies to grow multivirus-specific donorderived T-cells (mCTL), one from peripheral blood (PB) of adult CMV-seropositive donors and another from naive cord blood (CB). Using an adenoviral-vector expressing CMVpp65 or overlapping viral peptides for CMV (pp65 and IE-1), EBV (EBNA1 and LMP2), and Adenovirus (Hexon and Penton)
Abstracts / Biol Blood Marrow Transplant 22 (2016) S19eS481
presented to T cells by dendritic cells, monocytes, or EBV-LCL, we generated a single culture of mCTL. PB mCTL (Mean SFC:adeno:666, EBV:129, CMV:535) had more spot forming cells (SFC per 100,000 cells) (Mean, adeno: 666, EBV: 129, CMV: 535) than CB mCTL (adeno:117, EBV:95, CMV:67) by IFN-gamma ELISPOT assay but both contained cells specific for at least 1 virus. mCTL derived from both CB and PB contained a mixture of CD4+ and CD8+ T cells with an effector and central memory phenotype. Based on deep T cell receptor sequencing, CB mCTL were more polyclonal than PBderived mCTL. Thirteen patients were infused with PB mCTL and 12 patients with CB mCTL. Patients received CTL infusions from 35-384 days post transplant at a range of 5x106-2x107 cells/m2 with no toxicity or GvHD >grade II. We observed up to a 160-fold increase in virus-specific T-cells by 4 weeks post-CTL as measured by IFN-g ELISPOT assay. In 25 patients enrolled on these two studies, 16 patients experienced CMV, EBV, or adenovirus viral infections/reactivations before or immediately after mCTL infusion. Nine patients remained free of infection/reactivation. Eight of the patients had a complete response post-mCTL and 5 had a partial response, most coinciding with an increase in virus-specific T cells. Three patients did not respond to therapy. The overall the response rate in both groups was 81%. This study demonstrates that mCTL derived from the PB of seropositive donors as well as the CB of virus naive donors expand in vivo and are active against multiple viruses. Furthermore, by restoring immunity to multiple viruses simultaneously, the need for continued prophylaxis with pharmacotherapy is eliminated, thus, improving the efficiency and cost effectiveness of protecting SCT and CBT recipients from these potentially lethal viruses.
169 Targeting Wilms’ Tumor 1 Protein Following CD34-Selected Allografts By Adoptive Transfer of Donor-Derived CTLs in Patients with Advanced Multiple Myeloma Guenther Koehne 1, Satyajit Kosuri 1, Ekaterina S. Doubrovina 2, Evelyn Orlando 1, Sergio A. Giralt 1, Achim A. Jungbluth 3, Richard J. O’Reilly 4. 1 Department of Medicine, Adult Bone Marrow Transplant Service, Memorial Sloan Kettering Cancer Center, New York, NY; 2 Pediatrics, Memorial Sloan-Kettering Cancer Center, New York, NY; 3 Department of Pathology, Memorial Sloan Kettering Cancer Center, New York, NY; 4 Department of Pediatrics, Bone Marrow Transplant Service, Memorial Sloan Kettering Cancer Center, New York, NY Introduction: We have demonstrated the overexpression of Wilms’ tumor 1 (WT1) protein in myeloma cells by IHC and in HLA-A*0201+ pts by staining with a high-affinity fully human IgG1 mAb (ESK1) specific to the RMFPNAPYL/HLAA*0201 complex on malignant plasma cells. We report initial results from pts with plasma cell leukemia (PCL) or relapsed/ refractory multiple myeloma (rMM) who have been treated with CD34-selected allo transplants followed by the administration of donor-derived WT1-specific CTLs to induce an immunotherapeutic effect. Methods: In situ expression of WT1 was assessed by IHC analyses using a sequential double staining technique of MoAbs specific for CD138 and WT1. For staining with the RMFPNAPYL/HLA-A*0201 complex, BM samples were blocked with human FcR Blocking Reagent and then directly stained with MoAbs specific for CD38, CD56, CD45 and ESK1 or its isotype control human IgG1 and were analyzed by flow cytometry. WT1-specific CTLs were generated from the
S141
original stem cell donors by sensitization of CD3+ enriched Tcell fractions with autologous APCs loaded with the pool of overlapping pentadecapeptides of WT1. Following propagation, T-cells were harvested, counted and tested for antigen specific cytotoxicity, HLA-restriction, lack of alloreactivity and sterility. Pts received CD34-selected PBSC allografts after myeloablative cytoreduction with busulfan, melphalan and fludarabine. Pts were treated with 3 infusions of donorderived WT1-specific CTLs (5x10e6 cells/kg) starting 6 weeks post allo HSCT and 4 weekly thereafter. Results: Marrow from all pts with immunohistochemical documented plasma cell involvement stained positive for WT1 IHC while WT1 staining remained negative in pts in CR. Only pts expressing HLA-A*0201 that stained positively for WT1 by IHC also demonstrated expression of WT1 by the RMFPNAPYL/HLA-A*0201 complex, whereas pts lacking HLA-A*0201 but with active disease stained positive for WT1 IHC but not ESK1 staining. Seven pts, (3 PCL and 4 rMM), were treated with WT1-specific CTLs, 4 pts had persistent disease post CD34-selected allotransplant. Of these 4 pts 2 pts with PCL developed a striking rise of WT1-specific T-cell frequencies and developed a CR post WT1 CTL infusions lasting for up to 2 years. At 1 year post transplant all 3 pts with PCL were in CR; 2 pts with rMM achieved stable disease, 1 pt had a PR and 1 pt had died from progressive disease. Conclusion: Adoptive transfer of donor-derived WT1-specific CTLs following CD34-selected allografts in pts with PCL and rMM can achieve lasting remission underscoring the therapeutic potential of T cells specific for immunogenic WT1 peptides expressed on malignant plasma cells. Adverse events reported were consistent with those typically observed in this transplant population.
170 Allogeneic Umbilical Cord Blood Infusion for Adults with Ischemic Stroke Joanne Kurtzberg 1, Rebecca Durham 2, Daniel Laskowitz 3, Andrew E. Balber 4, Ellen Bennett 3. 1 The Carolinas Cord Blood Bank, Duke University Medical Center, Durham, NC; 2 Robertson Clinical and Translational Cell Therapy ProgramCT2, Duke University, Durham, NC; 3 Neurology, Duke University, Durham, NC; 4 The Carolinas Cord Blood Bank and Robertson Cell and Translational Therapy Program, Duke University Medical Center, Durham, NC Background: Approximately 87% of adult strokes are ischemic in etiology, and occur when cerebral blood flow is blocked by a clot or mechanical event. Immediately after the stroke, inflammatory mediators may exacerbate the development of cerebral edema and secondary tissue injury. Cell therapy may favorably alter the natural history of these processes through paracrine signaling that reduces inflammation, promotes angiogenesis, neurogenesis and recruitment of endogenous cell repair mechanisms. We hypothesize that cell therapy using non HLA matched umbilical cord blood (CB) from an unrelated donor would provide benefit by reducing the area of permanent injury and improving functional outcomes for these patients. Methods: The CoBIS study is an IRB approved, FDA IND sponsored, prospective, open-label, multi-center, Phase 1 safety study of a single intravenous infusion of allogeneic CB in 10 adults ages 18-80 years old. Cord blood units are selected by ethnicity, blood type, and ability to supply a dose of 0.5 e 1.5 x 107 TNCC/kg. Eligible patients include those experiencing a recent, acute cortical, hemispheric, ischemic stroke in the middle cerebral artery (MCA) distribution as detected by MRI as a diffusion weighted abnormality and
Abstracts / Biol Blood Marrow Transplant 22 (2016) S19eS481
Health, Bethesda, MD; 7 Experimental Transplantation and Immunology Branch, National Cancer Institute, NIH, Bethesda, MD; 8 Medical Oncology, Greenebaum Cancer Center, Baltimore, MD; 9 Georgia Regents University Cancer Center, Augusta, GA; 10 Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD; 11 Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA; 12 Translational Research Programs, Center for Cellular Immunotherapies, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA; 13 Experimental Transplantation and Immunology Branch, National Institutes of Health, National Cancer Institute, Bethesda, MD We previously demonstrated that pre-emptive DLI after low intensity HCT using rapamycin-resistant donor CD4+ T cells (12-day manufacturing; T-Rapa12) led to successful engraftment, GVL effects, and a moderate rate and severity of GVHD (NCT0074490; Fowler et al, Blood, 2013); subsequent patients received T-Rapa cells manufactured in 6 days (T-Rapa6). All patients received an HLA-matched sibling mobilized allograft after conditioning with fludarabine (total dose, 120 mg/m2) and cyclophosphamide (total dose, 1200 mg/m2). GVHD prophylaxis consisted of cyclosporine plus short-course sirolimus (through d14 post-HCT). Donor T-Rapa cells (2.5 x 107/kg) were given on d14 post-HCT. For the current analysis, chemotherapy responsive patients with lymphoid malignancies (n¼62) that received transplant and either T-Rapa12 or T-Rapa6 cell DLI were pooled for evaluation. Recipients were stratified into prognostic groups I/II (low-intermediate risk) versus III (high risk) based on histology and remission status as shown in Table 1 (after Kahl et al, Blood, 2007). In our cohorts, no significant differences were observed between Kahl I/II vs. Kahl III risk groups in regards to GVHD, chimerism, or overall survival (OS; Table 1). Estimated OS at 36-months post-HCT in Kahl I/II and Kahl III T-Rapa cohorts was 74.6% and 67.6%, respectively (p¼0.32 overall) (Fig. 1a). By comparison, in the Kahl et al study, which differed from our analysis in part by the inclusion of unrelated donors, estimated OS at 36-months was clearly divergent for Kahl I/II vs. Kahl III diagnoses (I: 60% II:55% III:26%; Fig. 1b). Our data suggest that the addition of a preemptive T-Rapa DLI may improve on previously reported overall survival results in low intensity HCT for high-risk lymphoid malignancies such as DLBCL (not in CR) and HD.
Table I
Number of Patients Age: Median (Range) Diagnoses
Acute GVHD all Grades (%) Acute GAVHD Grade 3/4 (%) Late Acute GVHD (%) Chronic GVHD (%) Day 100 Full* T Cell Chimerism (%) Day 100 Full* Myeloid Chimerism (%) 36 Month OS (%) *Donor Chimerism 96%
Kahl I/II
Kahl III
p value
24 59.5 (35-68) Follicular (9) Mantle Cell (11) DLBCL CR (2) CLL noCR (2)
N/A <0.0001 N/A
33.3 4.2 17.4 56.5 37.5
38 47 (22-66) DLBL noCR (22) Hodgkin’s (11) Gray Zone (3) PTCL noCR (3) 34.2 7.9 24.8 40.0 50.0
70.8
73.0
1.00 NS
74.6
67.6
0.32 NS
0.90 1.00 0.53 0.28 0.60
NS NS NS NS NS
Figure 1.
168 A PHASE 1 Perspective: Multivirus-Specific T CELLS from BOTH Cord Blood and BONE Marrow Transplant Donors Patrick J. Hanley 1, Michael Daniel Keller 2, Maria Martin Manso 3, Caridad Martinez 4, Kathryn Leung 4, Conrad Russell Y. Cruz 5, Cecilia Barese 3, Sarah McCormack 6, Min Luo 3, Robert A. Krance 4, David A. Jacobsohn 7, Cliona M. Rooney 8, Helen E. Heslop 8, Elizabeth J. Shpall 9, Catherine M. Bollard 2. 1 CETI, Blood and Marrow Transplantation, Children’s National Medical Center, Washington, DC; 2 Center for Cancer and Immunology Research, Children’s National Medical Center, Washington, DC; 3 CETI, CNMC, Washington, DC; 4 Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children’s Hospital Cancer Center, Methodist Hospital Houston, Houston, TX; 5 Children’s National Health System, Washington, DC; 6 Children’s National Medical Center, Washington, DC; 7 Division of Blood and Marrow Transplantation Center for Cancer and Blood Disorders, Children’s National Health System, Washington, DC; 8 Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children’s Hospital, Houston Methodist Hospital, Houston, TX; 9 Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX CMV, EBV and adenovirus are problematic in patients after stem cell (SCT) and cord blood transplantation (CBT) and are associated with morbidity and mortality. Deficiencies in conventional therapeutics have increased interest in an immunotherapeutic approach to viral disorders. We have developed 2 strategies to grow multivirus-specific donorderived T-cells (mCTL), one from peripheral blood (PB) of adult CMV-seropositive donors and another from naive cord blood (CB). Using an adenoviral-vector expressing CMVpp65 or overlapping viral peptides for CMV (pp65 and IE-1), EBV (EBNA1 and LMP2), and Adenovirus (Hexon and Penton)
Abstracts / Biol Blood Marrow Transplant 22 (2016) S19eS481
presented to T cells by dendritic cells, monocytes, or EBV-LCL, we generated a single culture of mCTL. PB mCTL (Mean SFC:adeno:666, EBV:129, CMV:535) had more spot forming cells (SFC per 100,000 cells) (Mean, adeno: 666, EBV: 129, CMV: 535) than CB mCTL (adeno:117, EBV:95, CMV:67) by IFN-gamma ELISPOT assay but both contained cells specific for at least 1 virus. mCTL derived from both CB and PB contained a mixture of CD4+ and CD8+ T cells with an effector and central memory phenotype. Based on deep T cell receptor sequencing, CB mCTL were more polyclonal than PBderived mCTL. Thirteen patients were infused with PB mCTL and 12 patients with CB mCTL. Patients received CTL infusions from 35-384 days post transplant at a range of 5x106-2x107 cells/m2 with no toxicity or GvHD >grade II. We observed up to a 160-fold increase in virus-specific T-cells by 4 weeks post-CTL as measured by IFN-g ELISPOT assay. In 25 patients enrolled on these two studies, 16 patients experienced CMV, EBV, or adenovirus viral infections/reactivations before or immediately after mCTL infusion. Nine patients remained free of infection/reactivation. Eight of the patients had a complete response post-mCTL and 5 had a partial response, most coinciding with an increase in virus-specific T cells. Three patients did not respond to therapy. The overall the response rate in both groups was 81%. This study demonstrates that mCTL derived from the PB of seropositive donors as well as the CB of virus naive donors expand in vivo and are active against multiple viruses. Furthermore, by restoring immunity to multiple viruses simultaneously, the need for continued prophylaxis with pharmacotherapy is eliminated, thus, improving the efficiency and cost effectiveness of protecting SCT and CBT recipients from these potentially lethal viruses.
169 Targeting Wilms’ Tumor 1 Protein Following CD34-Selected Allografts By Adoptive Transfer of Donor-Derived CTLs in Patients with Advanced Multiple Myeloma Guenther Koehne 1, Satyajit Kosuri 1, Ekaterina S. Doubrovina 2, Evelyn Orlando 1, Sergio A. Giralt 1, Achim A. Jungbluth 3, Richard J. O’Reilly 4. 1 Department of Medicine, Adult Bone Marrow Transplant Service, Memorial Sloan Kettering Cancer Center, New York, NY; 2 Pediatrics, Memorial Sloan-Kettering Cancer Center, New York, NY; 3 Department of Pathology, Memorial Sloan Kettering Cancer Center, New York, NY; 4 Department of Pediatrics, Bone Marrow Transplant Service, Memorial Sloan Kettering Cancer Center, New York, NY Introduction: We have demonstrated the overexpression of Wilms’ tumor 1 (WT1) protein in myeloma cells by IHC and in HLA-A*0201+ pts by staining with a high-affinity fully human IgG1 mAb (ESK1) specific to the RMFPNAPYL/HLAA*0201 complex on malignant plasma cells. We report initial results from pts with plasma cell leukemia (PCL) or relapsed/ refractory multiple myeloma (rMM) who have been treated with CD34-selected allo transplants followed by the administration of donor-derived WT1-specific CTLs to induce an immunotherapeutic effect. Methods: In situ expression of WT1 was assessed by IHC analyses using a sequential double staining technique of MoAbs specific for CD138 and WT1. For staining with the RMFPNAPYL/HLA-A*0201 complex, BM samples were blocked with human FcR Blocking Reagent and then directly stained with MoAbs specific for CD38, CD56, CD45 and ESK1 or its isotype control human IgG1 and were analyzed by flow cytometry. WT1-specific CTLs were generated from the
S141
original stem cell donors by sensitization of CD3+ enriched Tcell fractions with autologous APCs loaded with the pool of overlapping pentadecapeptides of WT1. Following propagation, T-cells were harvested, counted and tested for antigen specific cytotoxicity, HLA-restriction, lack of alloreactivity and sterility. Pts received CD34-selected PBSC allografts after myeloablative cytoreduction with busulfan, melphalan and fludarabine. Pts were treated with 3 infusions of donorderived WT1-specific CTLs (5x10e6 cells/kg) starting 6 weeks post allo HSCT and 4 weekly thereafter. Results: Marrow from all pts with immunohistochemical documented plasma cell involvement stained positive for WT1 IHC while WT1 staining remained negative in pts in CR. Only pts expressing HLA-A*0201 that stained positively for WT1 by IHC also demonstrated expression of WT1 by the RMFPNAPYL/HLA-A*0201 complex, whereas pts lacking HLA-A*0201 but with active disease stained positive for WT1 IHC but not ESK1 staining. Seven pts, (3 PCL and 4 rMM), were treated with WT1-specific CTLs, 4 pts had persistent disease post CD34-selected allotransplant. Of these 4 pts 2 pts with PCL developed a striking rise of WT1-specific T-cell frequencies and developed a CR post WT1 CTL infusions lasting for up to 2 years. At 1 year post transplant all 3 pts with PCL were in CR; 2 pts with rMM achieved stable disease, 1 pt had a PR and 1 pt had died from progressive disease. Conclusion: Adoptive transfer of donor-derived WT1-specific CTLs following CD34-selected allografts in pts with PCL and rMM can achieve lasting remission underscoring the therapeutic potential of T cells specific for immunogenic WT1 peptides expressed on malignant plasma cells. Adverse events reported were consistent with those typically observed in this transplant population.
170 Allogeneic Umbilical Cord Blood Infusion for Adults with Ischemic Stroke Joanne Kurtzberg 1, Rebecca Durham 2, Daniel Laskowitz 3, Andrew E. Balber 4, Ellen Bennett 3. 1 The Carolinas Cord Blood Bank, Duke University Medical Center, Durham, NC; 2 Robertson Clinical and Translational Cell Therapy ProgramCT2, Duke University, Durham, NC; 3 Neurology, Duke University, Durham, NC; 4 The Carolinas Cord Blood Bank and Robertson Cell and Translational Therapy Program, Duke University Medical Center, Durham, NC Background: Approximately 87% of adult strokes are ischemic in etiology, and occur when cerebral blood flow is blocked by a clot or mechanical event. Immediately after the stroke, inflammatory mediators may exacerbate the development of cerebral edema and secondary tissue injury. Cell therapy may favorably alter the natural history of these processes through paracrine signaling that reduces inflammation, promotes angiogenesis, neurogenesis and recruitment of endogenous cell repair mechanisms. We hypothesize that cell therapy using non HLA matched umbilical cord blood (CB) from an unrelated donor would provide benefit by reducing the area of permanent injury and improving functional outcomes for these patients. Methods: The CoBIS study is an IRB approved, FDA IND sponsored, prospective, open-label, multi-center, Phase 1 safety study of a single intravenous infusion of allogeneic CB in 10 adults ages 18-80 years old. Cord blood units are selected by ethnicity, blood type, and ability to supply a dose of 0.5 e 1.5 x 107 TNCC/kg. Eligible patients include those experiencing a recent, acute cortical, hemispheric, ischemic stroke in the middle cerebral artery (MCA) distribution as detected by MRI as a diffusion weighted abnormality and