Malcolm K Brenner

Publications (110)831.07 Total impact

• Article: Adoptive immunotherapy with allodepleted donor T-cells improves immune reconstitution after haploidentical stem cell transplantation.

  Persis J Amrolia, Giada Muccioli-Casadei, Helen Huls, Stuart Adams, April Durett, Adrian Gee, Eric Yvon, Heidi Weiss, Mark Cobbold, H Bobby Gaspar, Cliona Rooney, Ingrid Kuehnle, Victor Ghetie, John Schindler, Robert Krance, Helen E Heslop, Paul Veys, Ellen Vitetta, Malcolm K Brenner
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  ABSTRACT: Poor T lymphocyte reconstitution limits the use of haploidentical stem cell transplantation (SCT) because it results in a high mortality from viral infections. One approach to overcome this problem is to infuse donor T cells from which alloreactive lymphocytes have been selectively depleted, but the immunologic benefit of this approach is unknown. We have used an anti-CD25 immunotoxin to deplete alloreactive lymphocytes and have compared immune reconstitution after allodepleted donor T cells were infused at 2 dose levels into recipients of T-cell-depleted haploidentical SCT. Eight patients were treated at 10(4) cells/kg/dose, and 8 patients received 10(5) cells/kg/dose. Patients receiving 10(5) cells/kg/dose showed significantly improved T-cell recovery at 3, 4, and 5 months after SCT compared with those receiving 10(4) cells/kg/dose (P < .05). Accelerated T-cell recovery occurred as a result of expansion of the effector memory (CD45RA(-)CCR-7(-)) population (P < .05), suggesting that protective T-cell responses are likely to be long lived. T-cell-receptor signal joint excision circles (TRECs) were not detected in reconstituting T cells in dose-level 2 patients, indicating they are likely to be derived from the infused allodepleted cells. Spectratyping of the T cells at 4 months demonstrated a polyclonal Vbeta repertoire. Using tetramer and enzyme-linked immunospot (ELISPOT) assays, we have observed cytomegalovirus (CMV)- and Epstein-Barr virus (EBV)-specific responses in 4 of 6 evaluable patients at dose level 2 as early as 2 to 4 months after transplantation, whereas such responses were not observed until 6 to 12 months in dose-level 1 patients. The incidence of significant acute (2 of 16) and chronic graft-versus-host disease (GVHD; 2 of 15) was low. These data demonstrate that allodepleted donor T cells can be safely used to improve T-cell recovery after haploidentical SCT and may broaden the applicability of this approach.
  Blood 10/2006; 108(6):1797-808. · 9.90 Impact Factor
• Source

  Available from: Bjoern Chapuy

  Article: CD45 monoclonal antibody-mediated cytolysis of human NK and T lymphoma cells.

  Gerald G Wulf, Anita Boehnke, Bjoern Chapuy, Bertram Glass, Bernhard Hemmerlein, Roland Schroers, Malcolm K Brenner, Lorenz Truemper
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  ABSTRACT: The CD45 rat monoclonal IgG2b antibodies YTH24.5 and YTH54.12 act synergistically to produce cytolysis of normal lymphocytes and have been safely given to patients in conditioning regimens for allogeneic stem cell transplantation. The antibodies are not lytic for hematopoietic stem cells, but the depletion of the lymphoid lineage cells is profound and sustained. We evaluated the YTH24.5 and YTH54.12 pair for complement-dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), and apoptotic and antiproliferative effects against a panel of non-Hodgkin's lymphoma (NHL) cell lines and against primary specimens. Significant CDC activity was observed against two of two NK and one of four T lymphoma cell lines; moderate activity was seen against two of four T, and four of eight B lymphoma cell lines. In the responding cell lines, the lytic activity of YTH24.5 and YTH54.12 was as least as strong as that of alemtuzumab or antithymocyte globulin. The combination of YTH24.5 and YTH54.12 also induced ADCC in one of two NK and two of four T lymphoma cell lines, as well as three primary specimens, but was ineffective in B-NHL. The antibodies decreased viability in two of two NK and one lymphoma cell line, measurable as apoptosis or direct cell death in the cell lines NK92 and CEM, respectively. In a tumor model of Jurkat lymphoma in SCID mice, administration of YTH24.5 and YTH54.12 impaired local tumor growth and delayed systemic disease progression. CD45 antibodies YTH24.5 and YTH54.12 have lytic activity against NK and T lymphoma cells via CDC and ADCC, are effective in a preclinical tumor model, and may be candidates for immunotherapeutic approaches to the treatment of human NK and T cell lymphoma.
  Haematologica 08/2006; 91(7):886-94. · 6.42 Impact Factor
• Article: Immunotherapy of high-risk acute leukemia with a recipient (autologous) vaccine expressing transgenic human CD40L and IL-2 after chemotherapy and allogeneic stem cell transplantation.

  Raphaël F Rousseau, Ettore Biagi, Aurélie Dutour, Eric S Yvon, Michael P Brown, Tiffany Lin, Zhuyong Mei, Bambi Grilley, Edwina Popek, Helen E Heslop, Adrian P Gee, Robert A Krance, Uday Popat, George Carrum, Judith F Margolin, Malcolm K Brenner
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  ABSTRACT: CD40L generates immune responses in leukemia-bearing mice, an effect that is potentiated by IL-2. We studied the feasibility, safety, and immunologic efficacy of an IL-2- and CD40L-expressing recipient-derived tumor vaccine consisting of leukemic blasts admixed with skin fibroblasts transduced with adenoviral vectors encoding human IL-2 (hIL-2) and hCD40L. Ten patients (including 7 children) with high-risk acute myeloid (n = 4) or lymphoblastic (n = 6) leukemia in cytologic remission (after allogeneic stem cell transplantation [n = 9] or chemotherapy alone [n = 1]) received up to 6 subcutaneous injections of the IL-2/CD40L vaccine. None of the patients were receiving immunosuppressive drugs. No severe adverse reactions were noted. Immunization produced a 10- to 890-fold increase in the frequencies of major histocompatibility complex (MHC)-restricted T cells reactive against recipient-derived blasts. These leukemia-reactive T cells included both T-cytotoxic/T-helper 1 (Th1) and Th2 subclasses, as determined from their production of granzyme B, interferon-gamma, and interleukin-5. Two patients produced systemic IgG antibodies that bound to their blasts. Eight patients remained disease free for 27 to 62 months after treatment (5-year overall survival, 90%). Thus, even in heavily treated patients, including recipients of allogeneic stem cell transplants, recipient-derived antileukemia vaccines can induce immune responses reactive against leukemic blasts. This approach may be worthy of further study, particularly in patients with a high risk of relapse.
  Blood 03/2006; 107(4):1332-41. · 9.90 Impact Factor
• Article: Administration of latent membrane protein 2-specific cytotoxic T lymphocytes to patients with relapsed Epstein-Barr virus-positive lymphoma.

  Catherine M Bollard, M Helen Huls, Elizabeth Buza, Heidi Weiss, Vicky Torrano, Mary V Gresik, Jeff Chang, Adrian Gee, Stephen M Gottschalk, George Carrum, Malcolm K Brenner, Cliona M Rooney, Helen E Heslop
  Clinical Lymphoma & Myeloma 02/2006; 6(4):342-7. · 1.13 Impact Factor
• Article: Complement‐Fixing CD45 Monoclonal Antibodies to Facilitate Stem Cell Transplantation in Mouse and Man

  MALCOLM K. BRENNER, GERALD G. WULF, DONNA R. RILL, KANG-LI LUO, MARGARET A. GOODELL, ZUYONG MEI, INGRID KUEHNLE, MICHAEL P. BROWN, MARTIN PULÉ, HELEN E. HESLOP, ROBERT A. KRANCE
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  ABSTRACT: Broadening the applicability of stem cell therapies requires safer preparative regimens for patients. The CD45 antigen is present on all cells of the hematopoietic lineage, and using a murine model, we determined whether a lytic CD45 monoclonal antibody could produce persistent aplasia and whether it could facilitate syngeneic or allogeneic stem cell engraftment. After its systemic administration, we found that all leukocyte subsets in peripheral blood were markedly diminished, but only the effect on the lymphoid compartment was sustained and marrow progenitor cells were spared from destruction. Given the transient effects of the monoclonal antibody on myelopoiesis and the more persistent effects on lymphopoiesis, we asked whether this agent could contribute to donor hemopoietic engraftment after subablative transplantation. Treatment with anti-CD45 alone did not enhance syngeneic engraftment, consistent with its inability to destroy progenitor cells and permit competitive repopulation with syngeneic donor stem cells. By contrast, the combination of anti-CD45 and an otherwise inactive dose of total-body irradiation allowed engraftment of H2 fully allogeneic donor stem cells. We attribute this result to the recipient immunosuppression produced by depletion of CD45-positive lymphocytes. We next assessed a pair of unconjugated rat anti-human CD45 monoclonal antibodies (MAbs), YTH54.12 and YTH25.4, in a clinical trial in patients who were to receive stem cell transplantation for acute leukemia. The maximum tolerated dose of these MAbs, 400 μg/kg/day, produced a pattern of response identical to that seen in the mice, with marked reductions in circulating lymphoid and myeloid cells and sparing of early marrow progenitors. In two of three patients with active leukemia, the MAbs also produced a decrease in the percentage of leukemic blast cells in bone marrow. These pre-clinical and clinical results warrant further evaluation of anti-CD45 MAbs in subablative preparative regimens for stem cell transplantation.
  Annals of the New York Academy of Sciences 01/2006; 996(1):80 - 88. · 3.15 Impact Factor
• Article: A chimeric T cell antigen receptor that augments cytokine release and supports clonal expansion of primary human T cells.

  Martin A Pulè, Karin C Straathof, Gianpietro Dotti, Helen E Heslop, Cliona M Rooney, Malcolm K Brenner
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  ABSTRACT: The transduction of primary T cells to express chimeric T cell receptors (cTCR) for redirected targeting of tumor cells is an attractive strategy for generating tumor-specific T cells for adoptive therapy. However, tumor cells rarely provide costimulatory signals and hence cTCRs that transmit just a CD3zeta signal can only initiate target cell killing and interferon-gamma release and fail to induce full activation. Although incorporation of a CD28 component results in IL-2 release and limited proliferation, T cell activation remains incomplete. OX40 transmits a potent and prolonged T cell activation signal and is crucial for maintaining an immunological response. We hypothesize that the CD28-OX40-CD3zeta tripartite cytoplasmic domain will provide a full complement of activation, proliferation, and survival signals for enhanced anti-tumor activity.
  Molecular Therapy 12/2005; 12(5):933-41. · 6.87 Impact Factor
• Article: An inducible caspase 9 safety switch for T-cell therapy.

  Karin C Straathof, Martin A Pulè, Patricia Yotnda, Gianpietro Dotti, Elio F Vanin, Malcolm K Brenner, Helen E Heslop, David M Spencer, Cliona M Rooney
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  ABSTRACT: The efficacy of adoptive T-cell therapy as treatment for malignancies may be enhanced by genetic modification of infused cells. However, oncogenic events due to vector/transgene integration, and toxicities due to the infused cells themselves, have tempered enthusiasm. A safe and efficient means of removing aberrant cells in vivo would ameliorate these concerns. We describe a "safety switch" that can be stably and efficiently expressed in human T cells without impairing phenotype, function, or antigen specificity. This reagent is based on a modified human caspase 9 fused to a human FK506 binding protein (FKBP) to allow conditional dimerization using a small molecule pharmaceutical. A single 10-nM dose of synthetic dimerizer drug induces apoptosis in 99% of transduced cells selected for high transgene expression in vitro and in vivo. This system has several advantages over currently available suicide genes. First, it consists of human gene products with low potential immunogenicity. Second, administration of dimerizer drug has no effects other than the selective elimination of transduced T cells. Third, inducible caspase 9 maintains function in T cells overexpressing antiapoptotic molecules. These characteristics favor incorporation of inducible caspase 9 as a safety feature in human T-cell therapies.
  Blood 07/2005; 105(11):4247-54. · 9.90 Impact Factor
• Article: Human cytotoxic T lymphocytes with reduced sensitivity to Fas-induced apoptosis.

  Gianpietro Dotti, Barbara Savoldo, Martin Pule, Karin C Straathof, Ettore Biagi, Eric Yvon, Stephane Vigouroux, Malcolm K Brenner, Cliona M Rooney
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  ABSTRACT: Effector-memory T cells expressing Fas (Apo-1/CD95) are switched to an apoptotic program by cross-linking with Fas-ligand (FasL). Consequently, tumors that express FasL can induce apoptosis of infiltrating Fas-positive T lymphocytes and subdue any antitumor host immune response. Since Epstein-Barr virus (EBV)-associated tumors such as Hodgkin lymphoma (HL) and nasopharyngeal carcinoma (NPC) express FasL, we determined whether EBV-specific cytotoxic T lymphocytes (EBV-CTLs) could be modified to resist this evasion strategy. We show that long-term down-modulation of Fas can be achieved in EBV-CTLs by transduction with small interfering RNA (siRNA) encoded in a retrovirus. Modified T cells resisted Fas/FasL-mediated apoptosis compared with control cells and showed minimal cleavage of the caspase3 substrate poly(ADP-ribose) polymerase (PARP) protein after Fas engagement. Prolonged Fas stimulation selected a uniformly Fas(low) and FasL resistant population. Removal of responsiveness to this single death signal had no other discernible effects on EBV-CTLs. In particular, it did not lead to their autonomous growth since the modified EBV-CTLs remained polyclonal, and their survival and proliferation retained dependence on antigen-specific stimulation and on the presence of other physiologic growth signals. EBV-CTLs with knocked down Fas should have a selective functional and survival advantage over unmodified EBV-CTLs in the presence of tumors expressing FasL and may be of value for adoptive cellular therapy.
  Blood 07/2005; 105(12):4677-84. · 9.90 Impact Factor
• Article: Treatment of nasopharyngeal carcinoma with Epstein-Barr virus--specific T lymphocytes.

  Karin C M Straathof, Catherine M Bollard, Uday Popat, M Helen Huls, Teresita Lopez, M Craig Morriss, Mary V Gresik, Adrian P Gee, Heidi V Russell, Malcolm K Brenner, Cliona M Rooney, Helen E Heslop
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  ABSTRACT: Conventional treatment for nasopharyngeal carcinoma (NPC) frequently fails and is accompanied by severe long-term side effects. Since virtually all undifferentiated NPCs are associated with Epstein-Barr virus (EBV), this tumor is an attractive candidate for cellular immunotherapy targeted against tumor-associated viral antigens. We now demonstrate that EBV-specific cytotoxic T-cell (CTL) lines can readily be generated from individuals with NPC, notwithstanding the patients' prior exposure to chemotherapy/radiation. A total of 10 patients diagnosed with advanced NPC were treated with autologous CTLs. All patients tolerated the CTLs, although one developed increased swelling at the site of pre-existing disease. At 19 to 27 months after infusion, 4 patients treated in remission from locally advanced disease remain disease free. Of 6 patients with refractory disease prior to treatment, 2 had complete responses, and remain in remission over 11 to 23 months after treatment; 1 had a partial remission that persisted for 12 months; 1 has had stable disease for more than 14 months; and 2 had no response. These results demonstrate that administration of EBV-specific CTLs to patients with advanced NPC is feasible, appears to be safe, and can be associated with significant antitumor activity.
  Blood 04/2005; 105(5):1898-904. · 9.90 Impact Factor
• Source

  Available from: PubMed Central

  Article: Cytotoxic T lymphocyte therapy for Epstein-Barr virus+ Hodgkin's disease.

  Catherine M Bollard, Laura Aguilar, Karin C Straathof, Benedikt Gahn, M Helen Huls, Alexandra Rousseau, John Sixbey, M Victoria Gresik, George Carrum, Melissa Hudson, Dagmar Dilloo, Adrian Gee, Malcolm K Brenner, Cliona M Rooney, Helen E Heslop
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  ABSTRACT: Epstein Barr virus (EBV)+ Hodgkin's disease (HD) expresses clearly identified tumor antigens derived from the virus and could, in principle, be a target for adoptive immunotherapy with viral antigen-specific T cells. However, like most tumor-associated antigens in immunocompetent hosts, these potential targets are only weakly immunogenic, consisting primarily of the latent membrane protein (LMP)1 and LMP2 antigens. Moreover, Hodgkin tumors possess a range of tumor evasion strategies. Therefore, the likely value of immunotherapy with EBV-specific cytotoxic effector cells has been questioned. We have now used a combination of gene marking, tetramer, and functional analyses to track the fate and assess the activity of EBV cytotoxic T lymphocyte (CTL) lines administered to 14 patients treated for relapsed EBV+ HD. Gene marking studies showed that infused effector cells could further expand by several logs in vivo, contribute to the memory pool (persisting up to 12 mo), and traffic to tumor sites. Tetramer and functional analyses showed that T cells reactive with the tumor-associated antigen LMP2 were present in the infused lines, expanded in peripheral blood after infusion, and also entered tumor. Viral load decreased, demonstrating the biologic activity of the infused CTLs. Clinically, EBV CTLs were well tolerated, could control type B symptoms (fever, night sweats, and weight loss), and had antitumor activity. After CTL infusion, five patients were in complete remission at up to 40 mo, two of whom had clearly measurable tumor at the time of treatment. One additional patient had a partial response, and five had stable disease. The performance and fate of these human tumor antigen-specific T cells in vivo suggests that they might be of value for the treatment of EBV+ Hodgkin lymphoma.
  Journal of Experimental Medicine 01/2005; 200(12):1623-33. · 13.85 Impact Factor
• Article: Conserved CTL epitopes on the adenovirus hexon protein expand subgroup cross-reactive and subgroup-specific CD8+ T cells.

  Ann M Leen, Uluhan Sili, Elio F Vanin, Alan M Jewell, Weidong Xie, Dario Vignali, Pedro A Piedra, Malcolm K Brenner, Cliona M Rooney
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  ABSTRACT: Adenoviruses often cause lethal infections in immunocompromised individuals. Adoptive transfer of immune T cells offers a therapeutic option, but this strategy has been hindered by the paucity of information on molecular targets of cellular immunity and by the immunologic heterogeneity of the 51 human adenoviruses, which are grouped from A to F on the basis of genome size, composition, homology, and organization. Clonal analysis of the adenovirus-specific cytotoxic T lymphocyte (CTL) responses of seropositive individuals identified 5 novel CD8(+) T-cell epitopes, all located in conserved regions of the capsid protein hexon. Reactive T cells were cross-reactive between 2 to 4 groups, while no T cells specific for a single subgroup were detected. Thus, by exploiting these peptide targets, it is possible to prepare a T-cell population capable of reacting with most adenoviruses that cause disease in immunocompromised patients.
  Blood 11/2004; 104(8):2432-40. · 9.90 Impact Factor
• Article: Genetic modification of T lymphocytes for adoptive immunotherapy.

  Claudia Rossig, Malcolm K Brenner
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  ABSTRACT: Adoptive transfer of T lymphocytes is a promising therapy for malignancies-particularly of the hemopoietic system-and for otherwise intractable viral diseases. Efforts to broaden the approach have been limited by the physiology of the T cells themselves and by a range of immune evasion mechanisms developed by tumor cells. In this review we show how genetic modification of T cells is being used preclinically and in patients to overcome these limitations, by incorporation of novel receptors, resistance mechanisms, and control genes. We also discuss how the increasing safety and effectiveness of gene transfer technologies will lead to an increase in the use of gene-modified T cells for the treatment of a wider range of disorders.
  Molecular Therapy 08/2004; 10(1):5-18. · 6.87 Impact Factor
• Article: Antigen-induced regulatory T cells.

  Stephane Vigouroux, Eric Yvon, Ettore Biagi, Malcolm K Brenner
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  ABSTRACT: Regulatory T cells participate in immunologic homeostasis by active suppression of inappropriate immune responses. Regulatory T lymphocytes expressing CD4 and CD25 antigens and naturally present in the peripheral blood were the first to be phenotypically characterized. However, their small number and antigen nonspecific suppression has prompted efforts to identify and dissect antigen-specific regulatory T cells. In this review we discuss how antigen-specific regulatory T cells can be identified, the cellular and molecular mechanisms underlying their induction and activity, and the challenges facing their potential clinical application.
  Blood 08/2004; 104(1):26-33. · 9.90 Impact Factor
• Article: Haematopoietic stem cell transplantation for autoimmune disease: limits and future potential.

  Malcolm K Brenner
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  ABSTRACT: Stem cell transplantation (SCT) for autoimmune disease is handicapped by a lack of definitive clinical trials able to demonstrate an overall benefit. This deficiency will become more problematic as the impetus grows to introduce and evaluate additional technologies intended to improve the safety and efficacy of the procedure. The development of effective surrogate analyses to predict outcome by measuring resurgent autoimmune clones or by genomic- and proteomic-based technologies to detect early disease recurrence may be of value in assessing the benefits of these modifications without the need for full-scale, long-term, randomized trials. The introduction of safer allogeneic transplantation techniques may increase the effectiveness of the procedure, while work on marrow stem cell plasticity and/or fusion suggests that SCT may serve not simply to halt the autoimmune process, but also to contribute cells capable of healing or regenerating diseased organs. Finally, the introduction of therapeutic transgenes into transplanted cells may further increase the effectiveness of SCT, although the regulatory complexities of gene therapy trials will probably delay this process. All these innovations will ensure that the next decade will see major changes in the practice and purpose of SCT for autoimmune disease.
  Bailli&egrave re s Best Practice and Research in Clinical Haematology 07/2004; 17(2):359-74. · 2.64 Impact Factor
• Article: The generation and characterization of LMP2-specific CTLs for use as adoptive transfer from patients with relapsed EBV-positive Hodgkin disease.

  Catherine M Bollard, Karin C M Straathof, M Helen Huls, Alan Leen, Kristine Lacuesta, Alan Davis, Stephen Gottschalk, Malcolm K Brenner, Helen E Heslop, Cliona M Rooney
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  ABSTRACT: Cellular adoptive immunotherapy for virus-associated malignant disease is an attractive strategy, since viral antigens provide targets for specific T lymphocytes. In Epstein-Barr virus (EBV)-positive Hodgkin disease (HD), a limited number of EBV-encoded antigens such as the latent membrane antigens (LMP) 1 and 2 are expressed on the malignant Reed-Sternberg cells. The authors aimed to generate cytotoxic T lymphocytes (CTLs) from patients with relapsed HD by specifically targeting LMP2A. Patients with relapsed HD have highly immunosuppressive tumors and have been heavily pretreated with cytotoxic agents. As a result, monocytes and lymphocytes are numerically reduced and functionally impaired. Approaches using dendritic cells (DCs) as the sole antigen-presenting cell to expand LMP2-specific CTL lines in vitro have proved impractical. The authors now show how small amounts of patient peripheral blood can be used to produce DCs expressing LMP2 after Ad5F35 transduction, and how an initial reactivation of LMP2-specific CTLs can be followed by stimulation with lymphoblastoid cell lines overexpressing LMP2 from the same vector. Large numbers of LMP2-specific cytotoxic lymphocytes are produced that contain both CD4+ and CD8+ T cells (favoring long-term persistence in vivo) and recognize multiple LMP2 epitopes (minimizing the risk of tumor antigen loss variants). This approach is being used in a current clinical trial.
  Journla of Immunotherapy 06/2004; 27(4):317-27. · 3.27 Impact Factor
• Article: Prompt versus preemptive intervention for EBV lymphoproliferative disease.

  Hans-Joachim Wagner, Yee Chung Cheng, M Helen Huls, Adrian P Gee, Ingrid Kuehnle, Robert A Krance, Malcolm K Brenner, Cliona M Rooney, Helen E Heslop
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  ABSTRACT: Posttransplantation lymphoproliferative disorders (PTLDs) caused by uncontrolled expansion of Epstein-Barr virus (EBV)-infected B cells after hematopoietic stem cell transplantation (HSCT) can be predicted by an increase in EBV DNA in peripheral blood mononuclear cells. We used real-time quantitative polymerase chain reaction (RQ-PCR) analysis to determine whether frequent monitoring of EBV DNA to allow preemptive treatment is truly of value in patients after HSCT. More than 1300 samples from 85 recipients were analyzed. No patient with consistently low EBV DNA levels developed PTLD. Nine patients had a single episode with a high EBV load (more than 4000 EBV copies/microg peripheral blood mononuclear cell [PBMC] DNA), and 16 patients had high EBV loads detected on 2 or more occasions. Only 8 of these developed symptoms consistent with PTLD, and all were promptly and successfully treated with EBV-specific cytotoxic T cells or CD20 monoclonal antibody. Hence, quantitative measurement of EBV DNA may best be used to enable the prompt rather than the preemptive treatment of PTLD.
  Blood 06/2004; 103(10):3979-81. · 9.90 Impact Factor
• Source

  Available from: cytometry.com.cn

  Article: An in vivo propagated human acute myeloid leukemia expressing ABCA3.

  Kevin Norwood, Rui-Yu Wang, Charlotte Hirschmann-Jax, Michael Andreeff, Malcolm K Brenner, Margaret A Goodell, Gerald G Wulf
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  ABSTRACT: Analyzing the regenerative compartment in the blast cell population of patients with acute myeloid leukemia (AML) may yield important insights into the mechanisms of disease progression. Here we present findings with a human AML cell line (AML-SP1), initiated from leukemic precursor cells and consecutively propagated by serial xenotransplantation in vivo. AML-SP1 maintained the characteristics of a human AML, consistently exhibiting a small leukemic side population (SP) of blast cells with high Hoechst 33342 exclusion. In the AML-SP1 line, an increased expression of the ABC transporters MDR1, MRP, ABCG2 and ABCA3 was found in the SP cells. The detection of ABCA3 in leukemic progenitor cells merits further investigation with regard to intracellular drug transport in AML blast cells. In vivo propagation of leukemias, such as AML-SP1 is a model system of maintaining the populational heterogeneity of AML disease, especially the unique characteristics of leukemic SP cells.
  Leukemia Research 04/2004; 28(3):295-9. · 2.92 Impact Factor
• Article: Fiber-modified adenoviruses generate subgroup cross-reactive, adenovirus-specific cytotoxic T lymphocytes for therapeutic applications.

  Ann M Leen, Uluhan Sili, Barbara Savoldo, Alan M Jewell, Pedro A Piedra, Malcolm K Brenner, Cliona M Rooney
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  ABSTRACT: Adenovirus (Ad) infections are responsible for considerable morbidity and mortality, particularly in pediatric hematopoietic stem cell transplant (HSCT) recipients. To date there is no therapy. The present study was motivated by the potential for using adoptive immunotherapy as either prophylaxis or treatment for Ad infections and associated diseases. The authors have developed a protocol to reactivate Ad-specific memory T cells from peripheral blood mononuclear cells (PBMCs) using a clinical-grade adenoviral vector. Such lines contain a specific CD4 and CD8 T-cell component and are capable of recognizing and lysing target cells infected with wild-type Ad serotypes from different Ad groups. Furthermore, the frequency of Ad-specific precursors can be determined in PBMCs ex vivo and used as a means to assess changes in Ad-specific T-cell memory responses after infusion. This is the first report of a simple and reproducible method to activate and expand Ad-specific cytotoxic T lymphocytes (CTLs), which should be protective against the range of different Ad subtypes that affect transplant recipients.
  Blood 03/2004; 103(3):1011-9. · 9.90 Impact Factor
• Article: A strategy for treatment of Epstein-Barr virus-positive Hodgkin's disease by targeting interleukin 12 to the tumor environment using tumor antigen-specific T cells.

  Hans-Joachim Wagner, Catherine M Bollard, Stéphane Vigouroux, M Helen Huls, Robert Anderson, H Grant Prentice, Malcolm K Brenner, Helen E Heslop, Cliona M Rooney
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  ABSTRACT: Adoptive immunotherapy with Epstein-Barr virus (EBV)-specific cytotoxic T cells (CTL) is effective for the prophylaxis and treatment of EBV-induced lymphoma in hematopoietic stem cell recipients. However, in EBV-positive Hodgkin's disease (HD) the efficacy of adoptively transferred EBV-specific CTL may be limited by tumor-derived immunosuppressive factors, such as T-cell growth factor (TGF) beta, interleukin (IL)13 and the chemokine TARC. Local delivery of IL12 to tumor sites by tumor-specific CTL could provide direct antitumor effects and overcome the CTL-inhibitory effects of the Th2 tumor environment while avoiding the systemic toxicity of recombinant IL12. EBV-specific CTL transduced with a retrovirus vector expressing the p40 and p35 subunits of IL12 as a single molecule (Flexi-IL12), produced IL12 following antigenic stimulation. This resulted in an elevated production of Th1 cytokines, including interferon gamma and tumor necrosis factor alpha, and a reduction in the Th2 cytokines IL4 and IL5. Flexi-IL12-transduced CTL resisted the antiproliferative and anticytotoxic effects of exogenous TGFbeta, likely by antagonizing the TGFbeta-induced downregulation of the Th1 transcriptional factor T-bet. In addition, Flexi-IL12-transduced CTL demonstrated a proliferative advantage in the presence of inhibitory supernatants from HD-derived cell lines. Tumor-specific, Flexi-IL12-transduced EBV-specific CTL should have a functional advantage over unmodified CTL, particularly in the presence of the adverse Th2 cytokine environment produced by Hodgkin tumor cells.
  Cancer Gene Therapy 03/2004; 11(2):81-91. · 2.80 Impact Factor
• Source

  Available from: PubMed Central

  Article: Evidence for the presentation of major histocompatibility complex class I-restricted Epstein-Barr virus nuclear antigen 1 peptides to CD8+ T lymphocytes.

  Kui Shin Voo, Tihui Fu, Helen Y Wang, Judy Tellam, Helen E Heslop, Malcolm K Brenner, Cliona M Rooney, Rong-Fu Wang
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  ABSTRACT: The Epstein-Barr virus (EBV)-encoded nuclear antigen 1 (EBNA1) is expressed in all EBV-associated tumors, making it an important target for immunotherapy. However, evidence for major histocompatibility complex (MHC) class I-restricted EBNA1 peptides endogenously presented by EBV-transformed B and tumor cells remains elusive. Here we describe for the first time the identification of an endogenously processed human histocompatibility leukocyte antigen (HLA)-B8-restricted EBNA1 peptide that is recognized by CD8+ T cells. T cell recognition could be inhibited by the treatment of target cells with proteasome inhibitors that block the MHC class I antigen processing pathway, but not by an inhibitor (chloroquine) of MHC class II antigen processing. We also demonstrate that new protein synthesis is required for the generation of the HLA-B8 epitope for T cell recognition, suggesting that defective ribosomal products (DRiPs) are the major source of T cell epitopes. Experiments with protease inhibitors indicate that some serine proteases may participate in the degradation of EBNA1 DRiPs before they are further processed by proteasomes. These findings not only provide the first evidence of the presentation of an MHC class I-restricted EBNA1 epitope to CD8+ T cells, but also offer new insight into the molecular mechanisms involved in the processing and presentation of EBNA1.
  Journal of Experimental Medicine 03/2004; 199(4):459-70. · 13.85 Impact Factor