315 results on '"Kufer, P."'
Search Results
152. Behavioral Ecology and the Transition to Agriculture
- Author
-
KUFER, JOHANNA
- Published
- 2006
- Full Text
- View/download PDF
153. Evaluation Of CD33 Expression and Functional Analysis Of The CD33/CD3 Bispecific BiTE® Antibody AMG 330 In Primary AML Samples
- Author
-
Krupka, Christina, Kufer, Peter, Kischel, Roman, Zugmaier, Gerhard, Boegeholz, Jan, Köhnke, Thomas, Lichtenegger, Felix S, Schneider, Stephanie, Metzeler, Klaus H., Fiegl, Michael, Spiekermann, Karsten, Baeuerle, Patrick, Hiddemann, Wolfgang, Riethmüller, Gert, and Subklewe, Marion
- Abstract
Kufer: AMGEN Research (Munich) GmbH: Employment; AMGEN Inc.: Equity Ownership. Kischel:AMGEN Research (Munich) GmbH: Employment; AMGEN Inc.: Equity Ownership. Zugmaier:Amgen Research (Munich) GmbH: Employment; Amgen Inc.: Equity Ownership. Baeuerle:AMGEN Research (Munich) GmbH: Employment; AMGEN Inc.: Equity Ownership. Riethmüller:AMGEN Inc.: Equity Ownership.
- Published
- 2013
- Full Text
- View/download PDF
154. Blinatumomab Monotherapy Shows Efficacy in Patients with Relapsed Diffuse Large B Cell Lymphoma
- Author
-
Viardot, Andreas, Goebeler, Mariele, Noppeney, Richard, Krause, Stefan W., Kallert, Stefan, Ferstl, Barbara, Mackensen, Andreas, Rupertus, Kathrin, Soekler, Martin, Kanz, Lothar, Knop, Stefan, Topp, Max S., Scheele, Jurgen, Nagorsen, Dirk, Zugmaier, Gerhard, Degenhard, Evelyn, Schmidt, Margit, Kufer, Peter, Libicher, Martin, Einsele, Hermann, and Bargou, Ralf
- Abstract
Krause: Micromet: Research Funding. Mackensen:Micromet Inc.: Research Funding. Topp:Micromet: Consultancy, Honoraria. Scheele:Micromet Inc.: Employment, Equity Ownership, Patents & Royalties. Nagorsen:Micromet Inc.: Employment, Equity Ownership, Patents & Royalties. Zugmaier:Micromet: Employment. Degenhard:Micromet Inc: Employment. Schmidt:Micromet AG: Employment. Kufer:Micromet Inc: Employment, Equity Ownership. Libicher:Micromet Inc.: Consultancy, Honoraria. Bargou:Micromet: Consultancy, Honoraria.
- Published
- 2011
- Full Text
- View/download PDF
155. Treatment of Patients with Non-Hodgkin Lymphoma (NHL) with CD19/CD3 Bispecific Antibody Blinatumomab (MT103): Double-Step Dose Increase to Continuous Infusion of 60 μg/m2/d Is Tolerable and Highly Effective
- Author
-
Viardot, Andreas, Goebeler, Mariele, Scheele, Jurgen S., Zugmaier, Gerhard, Noppeney, Richard, Knop, Stefan, Topp, Max S, Nagorsen, Dirk, Klinger, Matthias, Schmidt, Margit, Klappers, Petra, Kufer, Peter, Einsele, Hermann, and Bargou, Ralf
- Abstract
Scheele: Micromet Inc.: Employment. Zugmaier:Micromet Inc.: Employment. Nagorsen:Micromet Inc.: Employment. Klinger:Micromet Inc.: Employment. Schmidt:Micromet Inc.: Employment. Klappers:Micromet. Inc: Employment. Kufer:Micromet Inc.: Employment. Bargou:Micromet Inc.: Consultancy.
- Published
- 2010
- Full Text
- View/download PDF
156. Autologous T Cells From AML Patients Can Be Effectively Recruited for In-Vitro Lysis of Blasts by a Novel CD33/CD3-Bispecific BiTE Antibody
- Author
-
Aigner, Michael, Feulner, Julian, Kischel, Roman, Kufer, Peter, Baeuerle, Patrick A, Mackensen, Andreas, and Krause, Stefan W.
- Abstract
Aigner: Micromet Inc.: Research Funding. Kischel:Micromet Inc.: Employment, Equity Ownership. Kufer:Micromet Inc.: Employment, Equity Ownership. Baeuerle:Micromet Inc.: Employment, Equity Ownership. Mackensen:Micromet. Inc.: Research Funding. Krause:Micromet Inc.: Research Funding.
- Published
- 2010
- Full Text
- View/download PDF
157. CD19/CD3 Bispecific Antibody Blinatumomab (MT-103) Is Highly Effective In Treatment of Patients with Minimal Residual Disease From Chemotherapy-Resistant B-Precursor Acute Lymphoblastic Leukemia
- Author
-
Topp, Max S, Zugmaier, Gerhard, Goekbuget, Nicola, Neumann, Svenja, Horst, Heinz-August, Raff, Thorsten, Brueggemann, Monika, Kneba, Michael, Viardot, Andreas, Schmid, Mathias, Schaich, Markus, Stelljes, Matthias, Goebeler, Mariele, Pfeiffer, Heike, Ottmann, Oliver, Burmeister, Thomas, Degenhard, Evelyn, Schmidt, Margit, Scheele, Jürgen, Einsele, Hermann, Kufer, Peter, Klinger, Matthias, Nagorsen, Dirk, Hoelzer, Dieter, and Bargou, Ralf
- Abstract
Zugmaier: Micromet Inc.: Employment. Degenhard:Micromet Inc.: Employment. Schmidt:Micromet Inc.: Employment. Scheele:Micromet Inc.: Employment. Kufer:Micromet Inc.: Employment. Klinger:Micromet Inc.: Employment. Nagorsen:Micromet Inc.: Employment. Bargou:Micromet Inc.: Consultancy.
- Published
- 2010
- Full Text
- View/download PDF
158. Report of a Phase II Trial of Single-Agent BiTE® Antibody Blinatumomab in Patients with Minimal Residual Disease (MRD) Positive B-Precursor Acute Lymphoblastic Leukemia (ALL).
- Author
-
Topp, Max S, Zugmaier, Gerhard, Goekbuget, Nicola, Kufer, Peter, Goebeler, Mariele, Klinger, Matthias, Degenhard, Evelyn, Baeuerle, Patrick A, Schmidt, Margit, Nagorsen, Dirk, Neumann, Svenja, Horst, Heinz A, Raff, Thorsten, Viardot, Andreas, Stelljes, Matthias, Schmid, Mathias, Ottmann, Oliver G., Burmeister, Thomas, Einsele, Hermann, Riethmueller, Gert, Hoelzer, Dieter, and Bargou, Ralf C
- Abstract
In patients with B-precursor ALL, presence of MRD after induction therapy or at any time point later predicts a hematological relapse, despite continued intensive chemotherapy or/and an allogeneic hematological stem cell transplantation (HSCT). Blinatumomab (MT103) targets the CD19 antigen, and is a member of a novel class of bispecific BiTE® antibodies that redirect T cells for lysis of target cells. A phase II study was conducted in collaboration with the German Multicenter Study Group on Adult Lymphoblastic Leukemia (GMALL) in patients with MRD-positive B precursor ALL.B–precursor ALL patients in complete hematological remission with either persistent or reappeared MRD at any time after consolidation I of front-line therapy were included. One treatment cycle of blinatumomab is a 4-week continuous i.v. infusion, which can be followed by allogeneic HSCT or in case of response by repeated consolidation cycles of blinatumomab with 2-week treatment-free intervals. The dose level at enrollment is 15 μg/m2/day. In patients, who do not respond within four cycles of treatment, the dose can be increased to 30 μg/m2/day. Molecular response is assessed by quantitative PCR of either individual rearrangements of immunoglobulin/TCR-genes or specific genetic aberrations such as bcr/abl or MLL-AF4.Nineteen patients have been treated to date and 16 patients are already evaluable for response. Thirteen of 16 evaluable patients went into molecular complete remission (CR) already after one cycle of blinatumomab. Three patients had a stable MRD level. Of note, 10 of the responding 13 patients had never achieved a molecular CR before blinatumomab treatment despite multiple treatment cycles including tyrosine kinase inhibitors in case of Ph-positive ALL. Two patients in molecular CR had an extra-medullary relapse one in testis and one in cerebro-spinal fluid, both representing immunological niches with limited accessibility for T cells. One patient with stable MRD level had a medullary relapse. All other patients are still relapse-free. None of the patients with molecular CR has shown a medullary relapse to date. The maximum follow-up of molecular CR has been 12 months. Most common adverse events (AEs) included lymphopenia, pyrexia, leucopenia and hypoimmunoglobulinemia. Only one patient had to be discontinued because of a fully reversible epileptical seizure. All other AEs resolved during treatment. Overall, treatment with blinatumomab was well tolerated. Response data of all 21 patients will be presented at ASH.Treatment with blinatumomab converted MRD-positive B–precursor ALL into molecular CR in 13 of 16 evaluable patients with refractory disease as indicated by persistent MRD after intensive chemotherapy. This amounts to a response rate of 81% providing thus the rationale for introducing blinatumomab as a novel agent in the treatment of B–precursor ALL.Zugmaier: Micromet: Employment, Equity Ownership. Goekbuget:Micromet: Consultancy, Research Funding. Kufer:Micromet: Employment, Equity Ownership, Patents & Royalties. Klinger:Micromet: Employment, Equity Ownership. Degenhard:Micromet: Employment, Equity Ownership. Baeuerle:Micromet: Employment, Equity Ownership. Schmidt:Micromet: Employment, Equity Ownership. Nagorsen:Micromet: Employment, Equity Ownership. Riethmueller:Micromet: Consultancy, Equity Ownership. Bargou:Micromet: Consultancy, Equity Ownership, Patents & Royalties.
- Published
- 2009
- Full Text
- View/download PDF
159. Transient Laboratory Findings Upon First Dosing with T-Cell Engaging BiTE® Antibody Blinatumomab in Non-Hodgkin Lymphoma Patients.
- Author
-
Nagorsen, Dirk, Zugmaier, Gerhard, Kufer, Peter, Klinger, Matthias, Viardot, Andreas, Goebeler, Mariele, Schmidt, Margit, Klappers, Petra, Wolf, Andreas, Brandl, Christian, Baeuerle, Patrick A, and Bargou, Ralf C
- Abstract
Nagorsen: Micromet: Employment, Equity Ownership. Zugmaier:Micromet: Employment, Equity Ownership. Kufer:Micromet: Employment, Equity Ownership, Patents & Royalties. Klinger:Micromet: Employment, Equity Ownership. Schmidt:Micromet: Employment, Equity Ownership. Klappers:Micromat: Employment, Equity Ownership. Wolf:Micromet: Employment, Equity Ownership. Brandl:Micromet: Employment, Equity Ownership. Baeuerle:Micromet: Employment, Equity Ownership. Bargou:Micromet: Consultancy, Patents & Royalties.
- Published
- 2009
- Full Text
- View/download PDF
160. Identification of a Predictive Factor for Reversible Neurological Adverse Events in a Subset of Non-Hodgkin Lymphoma Patients Treated with CD19-Specific BiTE ® Antibody Blinatumomab.
- Author
-
Zugmaier, Gerhard, Nagorsen, Dirk, Klinger, Matthias, Bargou, Ralf C, Goebeler, Mariele, Viardot, Andreas, Baeuerle, Patrick A, and Kufer, Peter
- Abstract
Zugmaier: Micromet: Employment, Equity Ownership. Nagorsen:Micromet: Employment, Equity Ownership. Klinger:Micromet: Employment, Equity Ownership. Bargou:Micromet: Consultancy, Patents & Royalties. Baeuerle:Micromet: Employment, Equity Ownership. Kufer:Micromet: Employment, Equity Ownership, Patents & Royalties.
- Published
- 2009
- Full Text
- View/download PDF
161. Confirmation of Safety, Efficacy and Response Duration in Non-Hodgkin Lymphoma Patients Treated with 60 μg/m2/d of BiTE® Antibody Blinatumomab.
- Author
-
Nagorsen, Dirk, Zugmaier, Gerhard, Viardot, Andreas, Goebeler, Mariele, Noppeney, Richard, Schmidt, Margit, Klappers, Petra, Baeuerle, Patrick A, Kufer, Peter, and Bargou, Ralf C
- Abstract
Nagorsen: Micromet: Employment, Equity Ownership. Zugmaier:Micromet: Employment, Equity Ownership. Schmidt:Micromet: Employment, Equity Ownership. Klappers:Micromet: Employment, Equity Ownership. Baeuerle:Micromet: Employment, Equity Ownership. Kufer:Micromet: Employment, Equity Ownership, Patents & Royalties. Bargou:Micromet: Consultancy, Patents & Royalties.
- Published
- 2009
- Full Text
- View/download PDF
162. Treatment with Anti-CD19 BiTE Antibody Blinatumomab (MT103 / MEDI-538) Is Able to Eliminate Minimal Residual Disease (MRD) in Patients with B-Precursor Acute Lymphoblastic Leukemia (ALL): First Results of An Ongoing Phase II Study.
- Author
-
Topp, Max, Goekbuget, Nikola, Kufer, Peter, Zugmaier, Gerhard, Degenhard, Evelyn, Neumann, Svenja, Horst, Heinz A, Viardot, Andreas, Schmid, Mathias, Ottmann, Oliver G., Schmidt, Margit, Reinhardt, Carsten, Baeuerle, Patrick A, Nagorsen, Dirk, Hoelzer, Dieter, and Bargou, Ralf C.
- Abstract
Blinatumomab is designed to link T cells with CD19-expressing target cells resulting in a non-restricted cytotoxic T-cell response and T-cell activation. Several preclinical studies showed that very low concentrations of blinatumomab mediate redirected lysis of human B lymphoma and leukemia cells. A phase I study has demonstrated significant clinical activity of the BiTE antibody in relapsed B-cell non-Hodgkin’s lymphoma (NHL). Based on these results, a phase II dose-escalating study was designed in collaboration with the German Multicenter Study Group on Adult Acute Lymphoblastic Leukemia to investigate efficacy, safety, and tolerability of blinatumomab in ALL patients who achieved a complete hematological remission, but remained MRD-positive. MRD is an independent prognostic factor that reflects primary drug resistance and is associated with a high relapse risk after start of consolidation. MRD was measured with standardized methods either by quantitative detection of individual rearrangements of immunoglobulin or T-cell receptor (TCR) rearrangements, or by bcr-abl fusion transcripts. The study population includes patients with acute B-precursor ALL at a minimum age of 18 years who show a bcr/abl signal above detection limit and/or at least one marker by rearrangement with a sensitivity of ≥10−4. Primary endpoint is the conversion rate to MRD negative status as defined by a bcr/abl signal below detection limit and/or by individual rearrangements of immunoglobulin or T-cell receptor (TCR) genes below 10−4. Secondary endpoints are time to hematological relapse, time to MRD progression, and time to molecular relapse. One treatment cycle of blinatumomab is a 4-week continuous intravenous infusion, which can be followed by allogeneic stem cell transplantation after the first cycle, or by repeated cycles after a 2-week treatment-free interval. MRD status is controlled after each treatment cycle. The starting dose level is 15 microgram/m2/24 hr, which may be escalated to 30 microgram/m2/24 hr and higher dose levels based on clinical activity and safety data. Four patients aged 31, 57, 62, and 65 years have so far been enrolled at the initial dose level of 15 microgram/m2/24 hr. Three out of the 4 patients had MRD by rearrangements at levels of 10−4, 10−3 and 10−1, and one patient had bcr/abl fusion transcripts at a level of 10−4. Three out of the 3 patients with rearrangements turned MRD negative after the first treatment cycle, independently from the level of MRD positivity at baseline. The other patient had stable bcr/abl level without signs of hematological relapse after the initial treatment cycle. Except for fever on the first 3 days of treatment, no clinically significant toxicities were recorded. Conclusion: These preliminary results indicate that treatment with blinatumomab is able to convert MRD positive ALL into an MRD negative status, and that this is well tolerated. Further exploration is warranted, as this may be a promising alternative treatment option especially for ALL refractory to chemotherapy.
- Published
- 2008
- Full Text
- View/download PDF
163. Sustained Response Duration Seen after Treatment with Single Agent Blinatumomab (MT103/MEDI-538) in the Ongoing Phase I Study MT103- 104 in Patients with Relapsed NHL
- Author
-
Bargou, Ralf C, Kufer, Peter, Goebeler, Mariele, Knop, Stefan, Einsele, Hermann, Noppeney, Richard, Viardot, Andreas, Baeuerle, Patrick A, Reinhardt, Carsten, Schmidt, Margit, Klappers, Petra, Nagorsen, Dirk, and Zugmaier, Gerhard
- Abstract
Introduction: Blinatumomab (MT103/MEDI-538), a BiTE antibody targeting the CD19 antigen, is a member of a novel class of molecules that redirect T cells for lysis of target cells. A Phase 1 dose escalation study is being conducted in patients with advanced NHL. Methods: Relapsed NHL patients requiring treatment were included. Most patients were pre-treated, with a median of 3 previous chemo/immunotherapy regimens. To date, 7 dose levels ranging from 0.0005 to 0.09 mg/m2/24 h have been tested. Blinatumomab was continuously infused as single agent over a period of 4–8 weeks. Objective responses were assessed by Cheson criteria and centrally reviewed. Results: To date 39 patients have been treated. Most common adverse events (AEs) included lymphopenia, pyrexia, and leukopenia. The majority of AEs (>95%) improved or resolved during treatment. Permanent treatment discontinuation due to AEs occurred in a total of 8 patients, of which 6 had fully reversible CNS events. One patient with a history of nearly fatal sepsis, pre-existing hypogammaglobulinemia and bone marrow affection by chemotherapy, experienced a fatal sepsis 5 weeks after treatment start. Dose-dependent activity was observed in mantle cell lymphoma, follicular lymphoma and CLL with responses observed in 11 out of 27 patients treated at doses of 0.015 mg/m2/24 h and higher. Five of those patients had complete and six had partial responses. At the dose level of 0.060 mg/m2/24 h, 7 out of 7 patients have shown objective responses. Beside one relapse after 14 months, no treatment failure has so far been observed for responders at dose levels of 0.030 mg/m2/24 h and 0.060 mg/m2/24 h. Five patients at these dose levels have ongoing responses for more than 6 months. Interestingly, partial remissions converted into complete remissions in two patients four weeks after end of infusion suggesting either reduction in lesion size due to efflux of a previously expanded T cell pool or prolonged T cell activity. Conclusions: Blinatumomab as single agent induced apparently durable responses in pre-treated B-NHL patients with the highest response rate at a dose level of 0.06 mg/m2/24 h. Recruitment is ongoing.
- Published
- 2008
- Full Text
- View/download PDF
164. The Anti-CD19 Bispecific T-Cell Engager (BiTE) MT103 (MEDI-538), Induces Dose-Dependent Complete and Partial Responses in Relapsed Non-Hodgkin Lymphoma (NHL): Phase I Study MT103-104.
- Author
-
Bargou, Ralf C., Kufer, Peter, Goebeler, Mariele, Knop, Stefan, Einsele, Hermann, Noppeney, Rudolf, Viardot, Andreas, Hess, Georg, Riethmueller, Gert, Reitsma, Dirk, Leo, Eugen, Reinhardt, Carsten, and Zugmaier, Gerhard
- Abstract
MT103 (MEDI-538), a bispecific, single chain construct specific for CD19 and CD3, is a member of a novel class of highly active antibody derivatives that direct T-cells to target cells. The primary objective of this study is to evaluate the safety and tolerability of a 4- to 8-week continuous intravenous infusion at increasing dose levels in patients with relapsed NHL, starting at dose level 1 (DL1) of 0.5μg/m2/24h. Dose-limiting toxicities (DLTs) were assessed during the first 2 weeks of infusion. To date, 31 patients have been included up to a dose level of 30μg/m2/24h. No DLTs were observed from DL1 to DL3 (5μg/m2/24h. At DL4 (5μg/m2/24h on day 1, followed by 15 μg/m2/24h maintenance dose), 7 patients (pts) were treated. One of the 7 pts had elevated liver enzymes up to CTC grade 3 after 2 weeks, and another patient showed reversible confusion, assessed as a DLT on the 2nd treatment day. For further optimization of safety and convenience of the initiation phase, the stepwise increase was modified to a constant infusion rate. This regimen was well tolerated and showed a favorable safety profile. Due to these modifications a total of 13 pts were treated at 15μg/m2/24h. At 30μg/m2/24h, 6 pts were treated; one DLT (metabolic acidosis, grade 4) was recorded. Confounding factors for this event were pre-existing renal insufficiency, fever and increasing anemia. The adverse events (AEs) were transient and the most common ones observed so far in all cohorts, regardless of causality, were (overall/grade 3 or 4) leucopenia (63%/37%), lymphopenia (59%/59%), pyrexia (55%/7%) elevated liver enzymes (44%/4%). At the dose level of 15μg/ m2/24h, MT103 induced 1 complete (CR) and 2 partial responses (PRs) in indolent lymphoma, which had not been observed at the previous dose levels. A total of 5 pts including the responders at this dose level had bone marrow involvement by indolent lymphoma. MT103 induced complete disappearance of lymphoma cells from the bone marrow in 3 and partial reduction in 2 of the 5 cases. One of these 5 patients had lymphoplasmocytoid lymphoma pre-treated with fludarabine. MT103 induced a 60% reduction of paraprotein. At the dose level of 30μg/m2/24h, clinical benefit was observed in mantle cell lymphoma (MCL), which had not been noted at the previous dose levels. One out of 3 evaluable pts with MCL enrolled at this dose level experienced a CR and one showed complete disappearance of lymphoma cells from the bone marrow. Best responses in the 26 evaluable pts overall were 2 CRs, 2 PRs, 2 minimal responses (MRs) and 13 SDs. Best responses in 14 evaluable patients starting at the dose level of 15μg/m2/24h were 2 CRs, 2 PRs, 2 MRs and 5 SDs.Conclusions: The current data suggest a favorable safety profile of MT103 administered as continuous 4 to 8 week infusion. MT103 induces CRs and PRs in relapsed indolent lymphoma and MCL in an apparently dose dependent fashion. Pronounced effects on lymphoma bone marrow infiltration have further demonstrated the clinical activity of MT103. Determination of the optimal biological dose is ongoing.
- Published
- 2007
- Full Text
- View/download PDF
165. T Cell Responses during Long-Term Continuous Infusion of MT103 (MEDI-538; Anti-CD19 BiTE) in Patients with Relapsed B-NHL: Data from Dose-Escalation Study MT103-104.
- Author
-
Klinger, Matthias, Kufer, Peter, Kirchinger, Petra, Lutterbüse, Ralf, Leo, Eugen, Reinhardt, Carsten, Bäuerle, Patrick, and Bargou, Ralf
- Abstract
MT103 (MEDI-538) is a bispecific single-chain antibody construct directed at CD3 on human T cells and CD19 on human B lymphoma and normal B cells. Transient linkage of B and T cells by MT103 provides T cells with a T cell receptor (TCR)-like signal leading to redirected lysis of B cell targets without apparent need of costimulation and inducing T cells to proliferate, secrete cytokines and upregulate surface activation markers. TCR-like signalling by MT103 is strictly dependent on the presence of target cells. Redirected lysis of CD19-positive cells by MT103 is seen at low picomolar concentrations and at low effector-to-target ratios. The in-vivo half-life of MT103 is approximately two hours. In the ongoing dose escalation study MT103-104, patients with relapsed B-NHL have so far received continuous infusion of MT103 at maintenance flow-rates of 0.5, 1.5, 5 and 15 μg/m2/24h for 4 or 8 weeks following a 3+3 dose escalation design. Serum concentrations of MT103 remained constant over the entire treatment period at a level depending on the respective maintenance flow-rate. Depletion of circulating B (lymphoma) cells could be observed more frequently with increasing dose levels (DL) from DL1 to DL3, and in all evaluable patients at DL4. Three of six evaluable patients at DL4 showed clinical responses (2 PR, 1 CR) according to standardized Cheson criteria, but no patient of DL1-3. The time courses of absolute CD4 and CD8 T cell counts in peripheral blood were determined by flow cytometry. CD8 T lymphocytes were further subdivided for analysis into naïve T cells, TCM (central memory T cells), TEM (effector memory T cells) and TEMRA (non-proliferating terminally differentiated CTL), and CD4 T lymphocytes into naïve T cells, TCM and TEM. Activation of CD4 and CD8 T cell subsets was determined by measuring upregulation of CD69, CD25 and HLA-DR. Serum levels of cytokines were determined as additional biomarkers for T cell activation. In 50% of patients at DL1 to DL3, CD4 and CD8 T cell counts increased during the course of treatment - over pre-treatment levels. The TEM subset from both CD4 and CD8 T cells accounted for most of the observed increases, while the naïve T cell subsets showed no increase but also no signs of apoptosis. The non-proliferative TEMRA subset of CD8 T cells also remained unchanged in most patients. This indicated that the selective increase of proliferation-competent TEM subsets was attributed to MT103-induced T cell proliferation. At DL4, all evaluable patients showed signs of T cell expansion after 2 weeks of MT103 infusion, which was most pronounced in those who developed a partial or complete remission. The increase of CD8 T cell counts was more pronounced than that of CD4 T cells. T cell expansion was accompanied by upregulation of T cell activation markers as well as by increases in serum concentrations of cytokines like IFN-γ. T cell expansion and activation reverted in all cases when the infusion of MT103 was stopped. In summary, MT103 induced a reversible secondary T cell response involving T cell activation and proliferation as well as T cell cytotoxicity against circulating B cells and lymphoma tissue. The dose-dependent T cell expansion observed during long-term infusion of MT103, particularly within the cytotoxic TEM subset of CD8 T cells, appears to play a key role for clinical activity.
- Published
- 2006
- Full Text
- View/download PDF
166. Activation of T Cells by Bispecific Single-Chain Construct MT103 (MEDI-538) Is Strictly Target Cell-Dependent.
- Author
-
Brischwein, Klaus, Hammond, Scott A., Parr, Larissa, Bernd, Schlereth, Locher, Mathias, Kufer, Peter, and Baeuerle, Patrick A.
- Abstract
Background: Bispecific antibodies have been extensively studied in vitro and in vivo for their use in redirected tumor cell lysis. A particular challenge of bispecific antibody constructs recognizing the CD3 signaling complex is to achieve a controlled polyclonal activation of T-cells that, ideally, is entirely dependent on the presence of target cells. If this is not the case, systemic production of inflammatory cytokines and secondary endothelial reactions may occur as side effects, as are observed with the murine anti-human CD3e antibody OKT-3 (muromab, Orthoclone®). Here we present evidence that MT103 (or MEDI-538), a bispecific single chain antibody of the BiTE class that targets CD19 and CD3, induces T-cell activation exclusively in the presence of target cells. Material and methods: Peripheral blood mononuclear cells from healthy donors were prepared by Ficoll density centrifugation. PBMC were incubated for 24 hours with MT103 in presence or absence of specific target cells. Target cell lysis was determined by measurement of adenylate kinase activity released from lysed cells. De novo expression of activation markers CD69 and CD25 on T-cells was assessed by flow cytometry using directly conjugated monoclonal antibodies, and the concentration of cytokines in the supernatant was determined by a commercial FACS-based bead array. Results: MT103 was analyzed for conditional T-cell activation. In the presence of target-expressing cell lines, low picomolar concentrations of MT103 were sufficient to stimulate a high percentage of peripheral human T-cells to express cytokines and surface activation markers, to enter into the cell cycle and to induce redirected lysis of target cells. However, in the absence of target cells, the BiTE molecules no longer detectably activated human T-cells even at concentrations exceeding the ED50 for redirected lysis and conditional T-cell activation by more than five orders of magnitude. Conclusion: Our data show that T-cell activation by MT103 is highly conditional in that it is strictly dependent on the presence.
- Published
- 2006
- Full Text
- View/download PDF
167. Relationship of T- and B-cell kinetics to clinical response in patients with relapsed/refractory non-Hodgkin lymphoma treated with blinatumomab.
- Author
-
Nägele V, Zugmaier G, Goebeler ME, Viardot A, Bargou R, Kufer P, and Klinger M
- Subjects
- B-Lymphocytes immunology, Humans, Lymphoma, Non-Hodgkin immunology, Neoplasm Recurrence, Local drug therapy, Neoplasm Recurrence, Local immunology, T-Lymphocytes immunology, Treatment Outcome, Antibodies, Bispecific therapeutic use, Antineoplastic Agents, Immunological therapeutic use, B-Lymphocytes drug effects, Lymphoma, Non-Hodgkin drug therapy, T-Lymphocytes drug effects
- Abstract
Blinatumomab is a first-in-class immunotherapy based on the bispecific T-cell engager (BiTE®) immune-oncology platform, which redirects CD3
+ T cells to kill CD19+ target cells. The objective of this analysis was to describe the correlation between B- and T-cell kinetics and response to blinatumomab in patients with relapsed or refractory (r/r) non-Hodgkin lymphoma (NHL). The clinical efficacy of treatment with blinatumomab in patients with r/r NHL was recently investigated in a phase 1 dose-escalation and expansion trial (NCT00274742) wherein 76 patients received blinatumomab by continuous intravenous infusion at various doses (0.5-90 μg/m2 /day). B-Cell depletion and expansion of CD3+ , CD4+ , and CD8+ T cells was analyzed in patients stratified per clinical response (complete response [CR], n = 16; partial response [PR], stable disease [SD], or progressive disease [PD], n = 54) for at least 4 weeks (additional 4 weeks after clinical benefit) from the date of administration of blinatumomab until dose-limiting toxicity or PD. B-cell depletion kinetics were faster in patients who had a CR than in patients who did not have a complete response (PR, SD, or PD). T-cell expansion (T-cell counts exceeding the baseline level on day 22) was more pronounced in patients with CR than in patients without CR. T-cell expansion in patients with CR correlated with increased T-cell counts of both CD4+ and CD8+ T cells compared with patients without CR. Patients with r/r NHL who achieved a CR had faster B-cell depletion and increased expansion of CD3+ , CD4+ , and CD8+ T cells than patients who did not achieve a CR., (Copyright © 2021 ISEH -- Society for Hematology and Stem Cells. Published by Elsevier Inc. All rights reserved.)- Published
- 2021
- Full Text
- View/download PDF
168. The PSMA-targeting Half-life Extended BiTE Therapy AMG 160 has Potent Antitumor Activity in Preclinical Models of Metastatic Castration-resistant Prostate Cancer.
- Author
-
Deegen P, Thomas O, Nolan-Stevaux O, Li S, Wahl J, Bogner P, Aeffner F, Friedrich M, Liao MZ, Matthes K, Rau D, Rattel B, Raum T, Kufer P, Coxon A, and Bailis JM
- Subjects
- Animals, CD3 Complex antagonists & inhibitors, CD3 Complex immunology, CD3 Complex metabolism, Cell Line, Tumor, Cytokines metabolism, Cytotoxicity, Immunologic, Disease Models, Animal, Dose-Response Relationship, Immunologic, Glutamate Carboxypeptidase II antagonists & inhibitors, Humans, Lymphocyte Activation immunology, Male, Mice, Prostatic Neoplasms, Castration-Resistant pathology, T-Lymphocytes metabolism, Treatment Outcome, Xenograft Model Antitumor Assays, Adoptive Transfer methods, Antigens, Surface immunology, Glutamate Carboxypeptidase II immunology, Prostatic Neoplasms, Castration-Resistant drug therapy, Prostatic Neoplasms, Castration-Resistant metabolism, T-Lymphocytes immunology
- Abstract
Purpose: Metastatic castration-resistant prostate cancer (mCRPC) remains a disease with high unmet medical need, as most patients do not achieve durable response with available treatments. Prostate-specific membrane antigen (PSMA) is a compelling target for mCRPC. It is highly expressed by primary and metastatic prostate cancer cells, with increased expression after progression on androgen deprivation therapy., Experimental Design: We developed AMG 160, a half-life extended, bispecific T-cell engager immuno-oncology therapy that binds PSMA on prostate cancer cells and cluster of differentiation 3 on T cells for treatment of mCRPC. AMG 160 was evaluated in vitro and in mCRPC xenograft models. AMG 160 tolerability was assessed in nonhuman primates (NHP). AMG 160 activity as monotherapy and in combination with a PSMA-imaging agent, novel hormonal therapy, and immune checkpoint blockade was evaluated., Results: AMG 160 induces potent, specific killing of PSMA-expressing prostate cancer cell lines in vitro , with half-maximal lysis of 6-42 pmol/L. In vivo , AMG 160 administered weekly at 0.2 mg/kg engages T cells administered systemically and promotes regression of established 22Rv-1 mCRPC xenograft tumors. AMG 160 is compatible with the imaging agent gallium 68-labeled PSMA-11, and shows enhanced cytotoxic activity when combined with enzalutamide or an anti-programmed death-1 antibody. AMG 160 exhibits an extended half-life and has an acceptable safety profile in NHPs., Conclusions: The preclinical characterization of AMG 160 highlights its potent antitumor activity in vitro and in vivo , and its potential for use with known diagnostic or therapeutic agents in mCRPC. These data support the ongoing clinical evaluation of AMG 160 in patients with mCRPC. See related commentary by Kamat et al., p. 2675 ., (©2021 American Association for Cancer Research.)
- Published
- 2021
- Full Text
- View/download PDF
169. Pasotuxizumab, a BiTE ® immune therapy for castration-resistant prostate cancer: Phase I, dose-escalation study findings.
- Author
-
Hummel HD, Kufer P, Grüllich C, Seggewiss-Bernhardt R, Deschler-Baier B, Chatterjee M, Goebeler ME, Miller K, de Santis M, Loidl W, Dittrich C, Buck A, Lapa C, Thurner A, Wittemer-Rump S, Koca G, Boix O, Döcke WD, Finnern R, Kusi H, Ajavon-Hartmann A, Stienen S, Sayehli CM, Polat B, and Bargou RC
- Subjects
- Aged, Aged, 80 and over, Humans, Male, Middle Aged, Antigens, Surface immunology, Biomarkers, Tumor blood, CD3 Complex immunology, Glutamate Carboxypeptidase II immunology, Immunotherapy, Infusions, Intravenous, Injections, Subcutaneous, Maximum Tolerated Dose, Treatment Outcome, Antibodies, Bispecific immunology, Antibodies, Bispecific pharmacokinetics, Antibodies, Bispecific therapeutic use, Antineoplastic Agents, Immunological immunology, Antineoplastic Agents, Immunological pharmacokinetics, Antineoplastic Agents, Immunological therapeutic use, Prostatic Neoplasms, Castration-Resistant blood, Prostatic Neoplasms, Castration-Resistant immunology, Prostatic Neoplasms, Castration-Resistant pathology, Prostatic Neoplasms, Castration-Resistant therapy
- Abstract
Aim: We report results of a first-in-human study of pasotuxizumab, a PSMA bispecific T-cell engager (BiTE
® ) immune therapy mediating T-cell killing of tumor cells in patients with advanced castration-resistant prostate cancer. Patients & methods: We assessed once-daily subcutaneous (SC) pasotuxizumab. All SC patients developed antidrug antibodies; therefore, continuous intravenous (cIV) infusion was assessed. Results: A total of 47 patients received pasotuxizumab (SC: n = 31, 0.5-172 μg/d; cIV: n = 16, 5-80 μg/d). The SC maximum tolerated dose was 172.0 μg/d. A sponsor change stopped the cIV cohort early; maximum tolerated dose was not determined. PSA responders occurred (>50% PSA decline: SC, n = 9; cIV, n = 3), including two long-term responders. Conclusion: Data support pasotuxizumab safety in advanced castration-resistant prostate cancer and represent evidence of BiTE monotherapy efficacy in solid tumors. Clinical trial registration: NCT01723475 (ClinicalTrials.gov).- Published
- 2021
- Full Text
- View/download PDF
170. The BiTE (bispecific T-cell engager) platform: Development and future potential of a targeted immuno-oncology therapy across tumor types.
- Author
-
Einsele H, Borghaei H, Orlowski RZ, Subklewe M, Roboz GJ, Zugmaier G, Kufer P, Iskander K, and Kantarjian HM
- Subjects
- Adolescent, Adult, Antigens, CD19 immunology, Antigens, Neoplasm immunology, B-Lymphocytes immunology, Child, Child, Preschool, Female, Humans, Male, Middle Aged, Treatment Outcome, Young Adult, Antibodies, Bispecific therapeutic use, Antineoplastic Agents therapeutic use, Hematologic Neoplasms therapy, Immunotherapy, Adoptive methods, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma therapy, Receptors, Chimeric Antigen immunology, T-Lymphocytes immunology
- Abstract
Immuno-oncology therapies engage the immune system to treat cancer. BiTE (bispecific T-cell engager) technology is a targeted immuno-oncology platform that connects patients' own T cells to malignant cells. The modular nature of BiTE technology facilitates the generation of molecules against tumor-specific antigens, allowing off-the-shelf immuno-oncotherapy. Blinatumomab was the first approved canonical BiTE molecule and targets CD19 surface antigens on B cells, making blinatumomab largely independent of genetic alterations or intracellular escape mechanisms. Additional BiTE molecules in development target other hematologic malignancies (eg, multiple myeloma, acute myeloid leukemia, and B-cell non-Hodgkin lymphoma) and solid tumors (eg, prostate cancer, glioblastoma, gastric cancer, and small-cell lung cancer). BiTE molecules with an extended half-life relative to the canonical BiTE molecules are also being developed. Advances in immuno-oncology made with BiTE technology could substantially improve the treatment of hematologic and solid tumors and offer enhanced activity in combination with other treatments., (© 2020 The Authors. Cancer published by Wiley Periodicals, Inc. on behalf of American Cancer Society.)
- Published
- 2020
- Full Text
- View/download PDF
171. Adhesion of T Cells to Endothelial Cells Facilitates Blinatumomab-Associated Neurologic Adverse Events.
- Author
-
Klinger M, Zugmaier G, Nägele V, Goebeler ME, Brandl C, Stelljes M, Lassmann H, von Stackelberg A, Bargou RC, and Kufer P
- Subjects
- Adult, B-Lymphocytes drug effects, B-Lymphocytes immunology, Brain blood supply, Brain immunology, Brain pathology, Cell Adhesion immunology, Cell Line, Child, Clinical Trials, Phase I as Topic, Clinical Trials, Phase II as Topic, Endothelial Cells drug effects, Endothelium, Vascular cytology, Endothelium, Vascular immunology, Endothelium, Vascular pathology, Female, Humans, Incidence, Lymphoma, Non-Hodgkin blood, Lymphoma, Non-Hodgkin drug therapy, Lymphoma, Non-Hodgkin immunology, Male, Microvessels cytology, Microvessels immunology, Microvessels pathology, Neoplasm Recurrence, Local drug therapy, Neoplasm Recurrence, Local immunology, Neurotoxicity Syndromes epidemiology, Neurotoxicity Syndromes pathology, Neurotoxicity Syndromes prevention & control, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma blood, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma drug therapy, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma immunology, T-Lymphocytes drug effects, Antibodies, Bispecific adverse effects, Cell Adhesion drug effects, Endothelial Cells immunology, Neurotoxicity Syndromes immunology, T-Lymphocytes immunology
- Abstract
Blinatumomab, a CD19/CD3-bispecific T-cell engager (BiTE) immuno-oncology therapy for the treatment of B-cell malignancies, is associated with neurologic adverse events in a subgroup of patients. Here, we provide evidence for a two-step process for the development of neurologic adverse events in response to blinatumomab: (i) blinatumomab induced B-cell-independent redistribution of peripheral T cells, including T-cell adhesion to blood vessel endothelium, endothelial activation, and T-cell transmigration into the perivascular space, where (ii) blinatumomab induced B-cell-dependent T-cell activation and cytokine release to potentially trigger neurologic adverse events. Evidence for this process includes (i) the coincidence of T-cell redistribution and the early occurrence of most neurologic adverse events, (ii) T-cell transmigration through brain microvascular endothelium, (iii) detection of T cells, B cells, and blinatumomab in cerebrospinal fluid, (iv) blinatumomab-induced T-cell rolling and adhesion to vascular endothelial cells in vitro , and (v) the ability of antiadhesive agents to interfere with blinatumomab-induced interactions between T cells and vascular endothelial cells in vitro and in patients. On the basis of these observations, we propose a model that could be the basis of mitigation strategies for neurologic adverse events associated with blinatumomab treatment and other T-cell therapies. SIGNIFICANCE: This study proposes T-cell adhesion to endothelial cells as a necessary but insufficient first step for development of blinatumomab-associated neurologic adverse events and suggests interfering with adhesion as a mitigation approach., (©2019 American Association for Cancer Research.)
- Published
- 2020
- Full Text
- View/download PDF
172. Antiviral Activity of HIV gp120-Targeting Bispecific T Cell Engager Antibody Constructs.
- Author
-
Brozy J, Schlaepfer E, Mueller CKS, Rochat MA, Rampini SK, Myburgh R, Raum T, Kufer P, Baeuerle PA, Muenz M, and Speck RF
- Subjects
- Animals, Antibodies, Bispecific immunology, CHO Cells, Cricetinae, Cricetulus, HIV Antibodies immunology, HIV Infections immunology, HIV Infections virology, HIV-1 drug effects, Humans, Immunotherapy, Leukocytes, Mononuclear immunology, Leukocytes, Mononuclear virology, Protein Binding, Virus Replication drug effects, Virus Replication immunology, Antibodies, Bispecific therapeutic use, Antiviral Agents therapeutic use, HIV Envelope Protein gp120 immunology, HIV Infections drug therapy, HIV-1 immunology, T-Lymphocytes immunology
- Abstract
Today's gold standard in HIV therapy is combined antiretroviral therapy (cART). It requires strict adherence by patients and lifelong medication, which can lower the viral load below detection limits and prevent HIV-associated immunodeficiency but cannot cure patients. The bispecific T cell-engaging (BiTE) antibody technology has demonstrated long-term relapse-free outcomes in patients with relapsed and refractory acute lymphocytic leukemia. Here, we generated BiTE antibody constructs that target the HIV-1 envelope protein gp120 (HIV gp120) using either the scFv B12 or VRC01, the first two extracellular domains (1 + 2) of human CD4 alone or joined to the single chain variable fragment (scFv) of the antibody 17b fused to an anti-human CD3ε scFv. These engineered human BiTE antibody constructs showed engagement of T cells for redirected lysis of HIV gp120-transfected CHO cells. Furthermore, they substantially inhibited HIV-1 replication in peripheral blood mononuclear cells (PBMCs) as well as in macrophages cocultured with autologous CD8
+ T cells, the most potent being the human CD4(1 + 2) BiTE [termed CD(1 + 2) h BiTE] antibody construct and the CD4(1 + 2)L17b BiTE antibody construct. The CD4(1 + 2) h BiTE antibody construct promoted HIV infection of human CD4- /CD8+ T cells. In contrast, the neutralizing B12 and the VRC01 BiTE antibody constructs, as well as the CD4(1 + 2)L17b BiTE antibody construct, did not. Thus, BiTE antibody constructs targeting HIV gp120 are very promising for constraining HIV and warrant further development as novel antiviral therapy with curative potential. IMPORTANCE HIV is a chronic infection well controlled with the current cART. However, we lack a cure for HIV, and the HIV pandemic goes on. Here, we showed in vitro and ex vivo that a BiTE antibody construct targeting HIV gp120 resulted in substantially reduced HIV replication. In addition, these BiTE antibody constructs display efficient killing of gp120-expressing cells and inhibited replication in ex vivo HIV-infected PBMCs or macrophages. We believe that BiTE antibody constructs recognizing HIV gp120 could be a very valuable strategy for a cure of HIV in combination with cART and compounds which reverse latency., (Copyright © 2018 American Society for Microbiology.)- Published
- 2018
- Full Text
- View/download PDF
173. Changes in clinical laboratory parameters and pharmacodynamic markers in response to blinatumomab treatment of patients with relapsed/refractory ALL.
- Author
-
Nägele V, Kratzer A, Zugmaier G, Holland C, Hijazi Y, Topp MS, Gökbuget N, Baeuerle PA, Kufer P, Wolf A, and Klinger M
- Abstract
Background: Blinatumomab has shown a remission rate of 69% in an exploratory single-arm, phase II dose-escalation study in adult patients with relapsed/refractory B-precursor acute lymphoblastic leukemia (ALL). We evaluated changes in laboratory parameters and immunopharmacodynamic markers in patients who received blinatumomab in the exploratory phase II study., Methods: Data from 36 adults with relapsed/refractory ALL receiving blinatumomab as 4-week continuous IV infusions in various dose cohorts were analyzed for changes in liver enzymes, first-dose parameters, peripheral blood cell subpopulations, and cytokine/granzyme B release. Associations with clinical response were evaluated., Results: Liver enzymes and inflammatory parameters transiently increased primarily during the first treatment week without clinical symptoms and reversed to baseline levels thereafter. B and T cells showed expected depletion and redistribution kinetics, respectively. Similarly, thrombocytes and T cells displayed an initial decline in cell counts, whereas neutrophils peaked during the first days after infusion start. T-cell redistribution coincided with upregulation of LFA-1 and CD69. Patients who responded to blinatumomab had more pronounced T-cell expansion, which was associated with proliferation of CD4
+ and CD8+ T cells and memory subsets. Release of cytokines and granzyme B primarily occurred during the first week of cycle 1, except for IL-10, which was released in subsequent cycles. Blinatumomab step-dosing was associated with lower cytokine release and lower body temperature., Conclusions: In this study of relapsed/refractory ALL, blinatumomab-induced changes in laboratory parameters were transient and reversible. The evaluated PD markers demonstrated blinatumomab activity, and the analysis of cytokines supported the rationale for stepwise dosing. ( ClinicalTrials.gov Identifier NCT01209286.).- Published
- 2017
- Full Text
- View/download PDF
174. Bispecific T cell engaging antibody constructs targeting a universally conserved part of the viral M2 ectodomain cure and prevent influenza A virus infection.
- Author
-
Pendzialek J, Roose K, Smet A, Schepens B, Kufer P, Raum T, Baeuerle PA, Muenz M, Saelens X, and Fiers W
- Subjects
- Animals, Antibodies, Bispecific administration & dosage, Antibodies, Monoclonal administration & dosage, Antibodies, Monoclonal, Humanized administration & dosage, Antibodies, Monoclonal, Humanized immunology, Antibodies, Viral blood, Cytotoxicity Tests, Immunologic, Immunologic Memory, Influenza Vaccines administration & dosage, Mice, Antibodies, Bispecific immunology, Influenza A virus chemistry, Influenza A virus immunology, Orthomyxoviridae Infections prevention & control, T-Lymphocytes immunology, Viral Matrix Proteins immunology
- Abstract
The ectodomain of the influenza A matrix protein 2 (M2e) is highly conserved amongst all influenza virus A subtypes. M2e is present on the surface of influenza A virus-infected cells, and therefore a suitable target for broadly protective therapies. We designed bispecific T cell engaging (BiTE
® ) antibody constructs specific for M2e by genetically fusing a single chain variable fragment (scFv) derived from an M2e-specific murine monoclonal antibody with a CD3ɛ-specific scFv. These so-called FLU BiTE® antibody constructs selectively mediate T cell dependent lysis of M2-expressing and influenza A virus infected cells and protect BALB/c mice against challenge with different influenza A virus subtypes. By humanizing the M2e-binding scFv, we generated human-like FLU BiTE® antibody constructs, with increased in vitro cytotoxic activity and in vivo protective capacity against influenza A virus infection. FLU BiTE® antibody constructs represent a promising new curative and prophylactic treatment option for influenza disease., (Copyright © 2017 Elsevier B.V. All rights reserved.)- Published
- 2017
- Full Text
- View/download PDF
175. Long-term relapse-free survival in a phase 2 study of blinatumomab for the treatment of patients with minimal residual disease in B-lineage acute lymphoblastic leukemia.
- Author
-
Gökbuget N, Zugmaier G, Klinger M, Kufer P, Stelljes M, Viardot A, Horst HA, Neumann S, Brüggemann M, Ottmann OG, Burmeister T, Wessiepe D, Topp MS, and Bargou R
- Subjects
- Adult, Aged, Follow-Up Studies, Fusion Proteins, bcr-abl genetics, Fusion Proteins, bcr-abl metabolism, Gene Expression, Humans, Middle Aged, Myeloid-Lymphoid Leukemia Protein genetics, Myeloid-Lymphoid Leukemia Protein metabolism, Neoplasm, Residual, Oncogene Proteins, Fusion genetics, Oncogene Proteins, Fusion metabolism, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma genetics, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma mortality, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma pathology, Recurrence, Survival Analysis, Transplantation, Homologous, Treatment Outcome, Antibodies, Bispecific therapeutic use, Antineoplastic Agents therapeutic use, Hematopoietic Stem Cell Transplantation, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma therapy
- Published
- 2017
- Full Text
- View/download PDF
176. A Threshold of Systemic MAGE-A Gene Expression Predicting Survival in Resected Non-Small Cell Lung Cancer.
- Author
-
Mecklenburg I, Sienel W, Schmid S, Passlick B, and Kufer P
- Subjects
- Aged, Carcinoma, Non-Small-Cell Lung pathology, Disease-Free Survival, Female, Gene Expression Regulation, Neoplastic, Humans, Kaplan-Meier Estimate, Male, Middle Aged, Neoplasm Metastasis, Neoplasm Recurrence, Local pathology, Biomarkers, Tumor blood, Carcinoma, Non-Small-Cell Lung blood, Melanoma-Specific Antigens blood, Neoplasm Recurrence, Local blood
- Abstract
Purpose: Quantitative measurement of minimal residual disease predicting recurrence in individual cancer patients is available only in very few indications, such as acute lymphoblastic leukemia, but is still missing in most solid tumors, including non-small cell lung cancer (NSCLC). Experimental Design: MAGE-A expression levels in blood and bone marrow determined as calibrator-normalized relative ratios by quantitative multimarker real-time RT-PCR for transcript amplification of MAGE -A1, -A2, -A3/6, -A4, -A10, and -A12 in 94 patients with completely resected NSCLC were correlated with survival in a clinical study. Results: Patients with MAGE-A expression levels ≥0.2 in at least one sample of bone marrow or blood at tumor surgery had a significantly reduced overall ( P = 0.007), cancer-free ( P = 0.002), and distant metastasis-free survival ( P < 0.001) versus patients below 0.2 in all samples without significant difference in locoregional recurrence-free survival. The corresponding HRs (≥0.2 vs. <0.2) for death, cancer-related death, and development of distant metastasis were 2.56 [95% confidence interval (CI), 1.42-4.63], 3.32 (95% CI, 1.66-6.61), and 4.03 (95% CI, 1.77-9.18), respectively. Five-year Kaplan-Meier estimates of distant metastasis-free survival were 43% (MAGE-A ≥ 0.2) versus 87% (MAGE-A < 0.2). Conclusions: MAGE-A expression in blood or bone marrow at tumor surgery is an independent predictor of survival in resected NSCLC. The reliable prediction of distant metastasis in individual patients with a statistically proven impact on overall survival may help to refine patient selection for adjuvant therapy urgently needed, especially in the clinical management of elderly patients. Clin Cancer Res; 23(5); 1213-9. ©2016 AACR ., (©2016 American Association for Cancer Research.)
- Published
- 2017
- Full Text
- View/download PDF
177. Impact of Diverse Immune Evasion Mechanisms of Cancer Cells on T Cells Engaged by EpCAM/CD3-Bispecific Antibody Construct AMG 110.
- Author
-
Deisting W, Raum T, Kufer P, Baeuerle PA, and Münz M
- Subjects
- Animals, Antibodies, Bispecific genetics, B7-H1 Antigen genetics, B7-H1 Antigen metabolism, CHO Cells, Cell Line, Tumor, Cricetulus, Cytotoxicity, Immunologic genetics, Cytotoxicity, Immunologic immunology, Epithelial Cell Adhesion Molecule, Gene Expression, Genetic Vectors genetics, Humans, Lymphocyte Activation genetics, Lymphocyte Activation immunology, Neoplasms genetics, Neoplasms metabolism, Proto-Oncogene Proteins c-bcl-2 genetics, Proto-Oncogene Proteins c-bcl-2 metabolism, Serpins genetics, Serpins metabolism, T-Lymphocytes metabolism, Transfection, Transforming Growth Factor beta genetics, Transforming Growth Factor beta metabolism, Antibodies, Bispecific immunology, Antigens, Neoplasm immunology, CD3 Complex immunology, Cell Adhesion Molecules immunology, Immune Evasion genetics, Neoplasms immunology, T-Lymphocytes immunology
- Abstract
Background: Bispecific T cell engager (BiTE®) are single-chain bispecific antibody constructs with dual specificity for CD3 on T cells and a surface antigen on target cells. They can elicit a polyclonal cytotoxic T cell response that is not restricted by T cell receptor (TCR) specificity, and surface expression of MHC class I/peptide antigen complexes. Using human EpCAM/CD3-bispecific BiTE® antibody construct AMG 110, we here assessed to what extent surface expression of PD-L1, cytoplasmic expression of indoleamine-2,3-deoxygenase type 1, Bcl-2 and serpin PI-9, and the presence of transforming growth factor beta (TGF-β), interleukin-10 (IL-10) and adenosine in culture medium can impact redirected lysis by AMG 110-engaged T cells., Methods: The seven factors, which are all involved in inhibiting T cell functions by cancer cells, were tested with human EpCAM-expressing Chinese hamster ovary (CHO) target cells at levels that in most cases exceeded those observed in a number of human cancer cell lines. Co-culture experiments were used to determine the impact of the evasion mechanisms on EC50 values and amplitude of redirected lysis by AMG 110, and on BiTE®-induced proliferation of previously resting human peripheral T cells., Findings: An inhibitory effect on redirected lysis by AMG 110-engaged T cells was seen upon overexpression of serpin PI-9, Bcl-2, TGF-β and PD-L1. An inhibitory effect on induction of T cell proliferation was only seen with CHO cells overexpressing IDO. In no case, a single evasion mechanism rendered target cells completely resistant to BiTE®-induced lysis, and even various combinations could not., Conclusions: Our data suggest that diverse mechanisms employed by cancer cells to fend off T cells cannot inactivate AMG 110-engaged T cells, and that inhibitory effects observed in vitro may be overcome by increased concentrations of the BiTE® antibody construct.
- Published
- 2015
- Full Text
- View/download PDF
178. Preclinical characterization of AMG 330, a CD3/CD33-bispecific T-cell-engaging antibody with potential for treatment of acute myelogenous leukemia.
- Author
-
Friedrich M, Henn A, Raum T, Bajtus M, Matthes K, Hendrich L, Wahl J, Hoffmann P, Kischel R, Kvesic M, Slootstra JW, Baeuerle PA, Kufer P, and Rattel B
- Subjects
- Animals, Antibodies, Monoclonal administration & dosage, Antibodies, Monoclonal immunology, Humans, Leukemia, Myeloid, Acute immunology, Leukemia, Myeloid, Acute pathology, Macaca fascicularis immunology, Mice, Molecular Targeted Therapy, Xenograft Model Antitumor Assays, Antibodies, Bispecific administration & dosage, CD3 Complex immunology, Leukemia, Myeloid, Acute drug therapy, Sialic Acid Binding Ig-like Lectin 3 immunology, T-Lymphocytes immunology
- Abstract
There is high demand for novel therapeutic options for patients with acute myelogenous leukemia (AML). One possible approach is the bispecific T-cell-engaging (BiTE, a registered trademark of Amgen) antibody AMG 330 with dual specificity for CD3 and the sialic acid-binding lectin CD33 (SIGLEC-3), which is frequently expressed on the surface of AML blasts and leukemic stem cells. AMG 330 binds with low nanomolar affinity to CD33 and CD3ε of both human and cynomolgus monkey origin. Eleven human AML cell lines expressing between 14,400 and 56,700 CD33 molecules per cell were all potently lysed with EC(50) values ranging between 0.4 pmol/L and 3 pmol/L (18-149 pg/mL) by previously resting, AMG 330-redirected T cells. Complete lysis was achieved after 40 hours of incubation. In the presence of AML cells, AMG 330 specifically induced expression of CD69 and CD25 as well as release of IFN-γ, TNF, interleukin (IL)-2, IL-10, and IL-6. Ex vivo, AMG 330 mediated autologous depletion of CD33-positive cells from cynomolgous monkey bone marrow aspirates. Soluble CD33 at concentrations found in bone marrow of patients with AML did not significantly affect activities of AMG 330. Neoexpression of CD33 on newly activated T cells was negligible as it was limited to 6% of T cells in only three out of ten human donors tested. Daily intravenous administration with as low as 0.002 mg/kg AMG 330 significantly prolonged survival of immunodeficient mice adoptively transferred with human MOLM-13 AML cells and human T cells. AMG 330 warrants further development as a potential therapy for AML., (©2014 American Association for Cancer Research.)
- Published
- 2014
- Full Text
- View/download PDF
179. Regression of human prostate cancer xenografts in mice by AMG 212/BAY2010112, a novel PSMA/CD3-Bispecific BiTE antibody cross-reactive with non-human primate antigens.
- Author
-
Friedrich M, Raum T, Lutterbuese R, Voelkel M, Deegen P, Rau D, Kischel R, Hoffmann P, Brandl C, Schuhmacher J, Mueller P, Finnern R, Fuergut M, Zopf D, Slootstra JW, Baeuerle PA, Rattel B, and Kufer P
- Subjects
- Animals, Antibodies, Bispecific genetics, Antibodies, Bispecific immunology, CD3 Complex genetics, CHO Cells, Cricetinae, Cross Reactions, Female, Haplorhini, Humans, Immunization, Passive methods, Macaca fascicularis, Male, Mice, Mice, Inbred BALB C, Mice, Inbred NOD, Mice, SCID, Prostatic Neoplasms pathology, Transfection, Xenograft Model Antitumor Assays, Antibodies, Bispecific pharmacology, Antigens, Surface immunology, CD3 Complex immunology, Glutamate Carboxypeptidase II immunology, Prostatic Neoplasms immunology, Prostatic Neoplasms therapy
- Abstract
For treatment of patients with prostate cancer (PCa), we developed a novel T cell-engaging (BiTE) antibody designated AMG 212 or BAY2010112 that is bispecific for prostate-specific membrane antigen (PSMA) and the CD3 epsilon subunit of the T cell receptor complex. AMG 212/BAY2010112 induced target cell-dependent activation and cytokine release of T cells, and efficiently redirected T cells for lysis of target cells. In addition to Chinese hamster ovary cells stably expressing human or cynomolgus monkey PSMA, T cells redirected by AMG 212/BAY2010112 also lysed human PCa cell lines VCaP, 22Rv1, MDA PCa 2b, C4-2, PC-3-huPSMA, and LnCaP at half maximal BiTE concentrations between 0.1 and 4 ng/mL (1.8-72 pmol/L). No lysis of PSMA-negative human PCa cell lines PC-3 and DU145 was observed. The subcutaneous (s.c.) formation of tumors from PC-3-huPSMA cells in NOD/SCID mice was significantly prevented by once daily intravenous (i.v.) injection of AMG 212/BAY2010112 at a dose level as low as 0.005 mg/kg/d. Rapid tumor shrinkage with complete remissions were observed in NOD/SCID mice bearing established s.c. 22Rv1 xenografts after repeated daily treatment with AMG 212/BAY2010112 by either the i.v. or s.c. route. Of note, 22Rv1 tumors were grown in the absence of human T cells followed by intraperitoneal injection of T cells 3 days before BiTE treatment. No effects on tumor growth were observed in the absence of human T cells or AMG 212/BAY2010112. On the basis of these preclinical results, AMG 212/BAY2010112 appears as a promising new BiTE antibody for the treatment of patients with PSMA-expressing PCa.
- Published
- 2012
- Full Text
- View/download PDF
180. Blinatumomab: a historical perspective.
- Author
-
Nagorsen D, Kufer P, Baeuerle PA, and Bargou R
- Subjects
- Animals, Antibodies, Bispecific administration & dosage, Antibodies, Bispecific chemistry, Clinical Trials as Topic, Humans, Lymphoma, Large B-Cell, Diffuse drug therapy, Lymphoma, Mantle-Cell drug therapy, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma drug therapy, Antibodies, Bispecific therapeutic use
- Abstract
For decades, chemotherapy has been the backbone for the treatment of patients with B cell malignancies. Depending on the individual disease, monoclonal antibodies, irradiation and/or hematopoietic stem cell transplantation are added. However, the current standard of care--particularly for patients with relapsed disease--is often not sufficient to achieve durable remissions. A highly promising new drug candidate in late-stage clinical development for treatment of B cell malignancies is blinatumomab (MT103 or AMG 103). This bispecific antibody construct has dual specificity for CD19 and CD3 and belongs to the class of bispecific T cell engager (BiTE®) antibodies, which can potentially engage all cytotoxic T cells of a patient for redirected lysis of tumor cells. Here, we review how blinatumomab has so far been pre-clinically and clinically developed for the treatment of patients with non-Hodgkin's lymphoma and acute lymphoblastic leukemia., (Copyright © 2012 Elsevier Inc. All rights reserved.)
- Published
- 2012
- Full Text
- View/download PDF
181. Targeted therapy with the T-cell-engaging antibody blinatumomab of chemotherapy-refractory minimal residual disease in B-lineage acute lymphoblastic leukemia patients results in high response rate and prolonged leukemia-free survival.
- Author
-
Topp MS, Kufer P, Gökbuget N, Goebeler M, Klinger M, Neumann S, Horst HA, Raff T, Viardot A, Schmid M, Stelljes M, Schaich M, Degenhard E, Köhne-Volland R, Brüggemann M, Ottmann O, Pfeifer H, Burmeister T, Nagorsen D, Schmidt M, Lutterbuese R, Reinhardt C, Baeuerle PA, Kneba M, Einsele H, Riethmüller G, Hoelzer D, Zugmaier G, and Bargou RC
- Subjects
- Adult, Agammaglobulinemia chemically induced, Aged, Aged, 80 and over, Antibodies, Bispecific adverse effects, Antibodies, Bispecific pharmacokinetics, Antibody Specificity, Antigens, CD19 immunology, Antigens, Neoplasm immunology, B-Lymphocytes immunology, B-Lymphocytes pathology, Bone Marrow pathology, CD3 Complex immunology, Cell Lineage, Combined Modality Therapy, Disease-Free Survival, Drug Resistance, Neoplasm, Female, Humans, Kaplan-Meier Estimate, Lymphocyte Activation, Lymphopenia chemically induced, Male, Middle Aged, Molecular Targeted Therapy, Neoplasm, Residual, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma drug therapy, Remission Induction, T-Cell Antigen Receptor Specificity drug effects, T-Cell Antigen Receptor Specificity immunology, T-Lymphocytes, Cytotoxic immunology, Young Adult, Antibodies, Bispecific therapeutic use, Drug Delivery Systems, Immunotherapy methods, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma therapy, T-Lymphocytes, Cytotoxic drug effects
- Abstract
Purpose: Blinatumomab, a bispecific single-chain antibody targeting the CD19 antigen, is a member of a novel class of antibodies that redirect T cells for selective lysis of tumor cells. In acute lymphoblastic leukemia (ALL), persistence or relapse of minimal residual disease (MRD) after chemotherapy indicates resistance to chemotherapy and results in hematologic relapse. A phase II clinical study was conducted to determine the efficacy of blinatumomab in MRD-positive B-lineage ALL., Patients and Methods: Patients with MRD persistence or relapse after induction and consolidation therapy were included. MRD was assessed by quantitative reverse transcriptase polymerase chain reaction for either rearrangements of immunoglobulin or T-cell receptor genes, or specific genetic aberrations. Blinatumomab was administered as a 4-week continuous intravenous infusion at a dose of 15 μg/m2/24 hours., Results: Twenty-one patients were treated, of whom 16 patients became MRD negative. One patient was not evaluable due to a grade 3 adverse event leading to treatment discontinuation. Among the 16 responders, 12 patients had been molecularly refractory to previous chemotherapy. Probability for relapse-free survival is 78% at a median follow-up of 405 days. The most frequent grade 3 and 4 adverse event was lymphopenia, which was completely reversible like most other adverse events., Conclusion: Blinatumomab is an efficacious and well-tolerated treatment in patients with MRD-positive B-lineage ALL after intensive chemotherapy. T cells engaged by blinatumomab seem capable of eradicating chemotherapy-resistant tumor cells that otherwise cause clinical relapse.
- Published
- 2011
- Full Text
- View/download PDF
182. Side-by-side analysis of five clinically tested anti-EpCAM monoclonal antibodies.
- Author
-
Münz M, Murr A, Kvesic M, Rau D, Mangold S, Pflanz S, Lumsden J, Volkland J, Fagerberg J, Riethmüller G, Rüttinger D, Kufer P, Baeuerle PA, and Raum T
- Abstract
Background: Epithelial cell adhesion molecule (EpCAM) is frequently and highly expressed on human carcinomas. The emerging role of EpCAM as a signalling receptor and activator of the wnt pathway, and its expression on tumor-initiating cells, further add to its attractiveness as target for immunotherapy of cancer. Thus far, five conventional monoclonal IgG antibodies have been tested in cancer patients. These are murine IgG2a edrecolomab and its murine/human chimeric IgG1 antibody version, and humanized, human-engineered and fully human IgG1 antibodies 3622W94, ING-1, and adecatumumab (MT201), respectively. Here we compared all anti-EpCAM antibodies in an attempt to explain differences in clinical activity and safety., Methods: We recombinantly produced all antibodies but murine edrecolomab and investigated them for binding affinity, EpCAM epitope recognition, ADCC and CDC, and inhibition of breast cancer cell proliferation., Results: ING-1 and 3622W94 bound to EpCAM with much higher affinity than adecatumumab and edrecolomab. Edrecolomab, ING-1, and 3622W94 all recognized epitopes in the exon 2-encoded N-terminal domain of EpCAM, while adecatumumab recognized a more membrane proximal epitope encoded by exon 5. All antibodies induced lysis of EpCAM-expressing cancer cell lines by both ADCC and CDC with potencies that correlated with their binding affinities. The chimeric version of edrecolomab with a human Fcγ1 domain was much more potent in ADCC than the murine IgG2a version. Only adecatumumab showed a significant inhibition of MCF-7 breast cancer cell proliferation in the absence of complement and immune cells., Conclusion: A moderate binding affinity and recognition of a distinct domain of EpCAM may best explain why adecatumumab showed a larger therapeutic window in cancer patients than the two high-affinity IgG1 antibodies ING-1 and 3622W94, both of which caused acute pancreatitis.
- Published
- 2010
- Full Text
- View/download PDF
183. Highly efficient elimination of colorectal tumor-initiating cells by an EpCAM/CD3-bispecific antibody engaging human T cells.
- Author
-
Herrmann I, Baeuerle PA, Friedrich M, Murr A, Filusch S, Rüttinger D, Majdoub MW, Sharma S, Kufer P, Raum T, and Münz M
- Subjects
- Animals, Colorectal Neoplasms pathology, Epithelial Cell Adhesion Molecule, Flow Cytometry, Humans, Mice, Mice, Inbred NOD, Mice, SCID, Antibodies, Bispecific immunology, Antigens, Neoplasm immunology, CD3 Complex immunology, Cell Adhesion Molecules immunology, Colorectal Neoplasms immunology, T-Lymphocytes immunology
- Abstract
With their resistance to genotoxic and anti-proliferative drugs and potential to grow tumors and metastases from very few cells, cancer stem or tumor-initiating cells (TICs) are a severe limitation for the treatment of cancer by conventional therapies. Here, we explored whether human T cells that are redirected via an EpCAM/CD3-bispecific antibody called MT110 can lyse colorectal TICs and prevent tumor growth from TICs. MT110 recognizes EpCAM, a cell adhesion molecule expressed on TICs from diverse human carcinoma, which was recently shown to promote tumor growth through engagement of elements of the wnt pathway. MT110 was highly potent in mediating complete redirected lysis of KRAS-, PI3 kinase- and BRAF-mutated colorectal TICs, as demonstrated in a soft agar assay. In immunodeficient mice, MT110 prevented growth of tumors from a 5,000-fold excess of a minimally tumorigenic TIC dose. T cells engaged by MT110 may provide a potent therapeutic means to eradicate TICs and bulk tumor cells derived thereof.
- Published
- 2010
- Full Text
- View/download PDF
184. T cell-engaging BiTE antibodies specific for EGFR potently eliminate KRAS- and BRAF-mutated colorectal cancer cells.
- Author
-
Lutterbuese R, Raum T, Kischel R, Hoffmann P, Mangold S, Rattel B, Friedrich M, Thomas O, Lorenczewski G, Rau D, Schaller E, Herrmann I, Wolf A, Urbig T, Baeuerle PA, and Kufer P
- Subjects
- Antibodies, Monoclonal, Antibodies, Monoclonal, Humanized, Cetuximab, Colorectal Neoplasms pathology, ErbB Receptors genetics, ErbB Receptors metabolism, Genes, ras drug effects, Humans, Mutation drug effects, Neoplasms genetics, Proto-Oncogene Proteins B-raf, Risk Factors, Colorectal Neoplasms drug therapy, Colorectal Neoplasms genetics, ErbB Receptors antagonists & inhibitors
- Abstract
Epidermal growth factor receptor (EGFR)-specific monoclonal antibodies predominantly inhibit colorectal cancer (CRC) growth by interfering with receptor signaling. Recent analyses have shown that patients with CRC with mutated KRAS and BRAF oncogenes do not profit from treatment with such antibodies. Here we have used the binding domains of cetuximab and pantitumumab for constructing T cell-engaging BiTE antibodies. Both EGFR-specific BiTE antibodies mediated potent redirected lysis of KRAS- and BRAF-mutated CRC lines by human T cells at subpicomolar concentrations. The cetuximab-based BiTE antibody also prevented at very low doses growth of tumors from KRAS- and BRAF-mutated human CRC xenografts, whereas cetuximab was not effective. In nonhuman primates, no significant adverse events were observed during treatment for 3 wk at BiTE serum concentrations inducing, within 1 d, complete lysis of EGFR-overexpressing cancer cells. EGFR-specific BiTE antibodies may have potential to treat CRC that does not respond to conventional antibodies.
- Published
- 2010
- Full Text
- View/download PDF
185. Antibody recognition of a unique tumor-specific glycopeptide antigen.
- Author
-
Brooks CL, Schietinger A, Borisova SN, Kufer P, Okon M, Hirama T, Mackenzie CR, Wang LX, Schreiber H, and Evans SV
- Subjects
- Animals, Antibodies, Monoclonal, Antibody Affinity, Antibody Specificity, Antigen-Antibody Complex chemistry, Crystallography, X-Ray, Epitopes chemistry, Glycopeptides chemistry, Glycopeptides immunology, Humans, Immunoglobulin Fab Fragments chemistry, In Vitro Techniques, Mice, Models, Molecular, Nuclear Magnetic Resonance, Biomolecular, Protein Conformation, Static Electricity, Surface Plasmon Resonance, Antigens, Tumor-Associated, Carbohydrate chemistry
- Abstract
Aberrant glycosylation and the overexpression of certain carbohydrate moieties is a consistent feature of cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy. One of the most common aberrations in glycosylation patterns is the presentation of a single O-linked N-acetylgalactosamine on a threonine or serine residue known as the "Tn antigen." Whereas the ubiquitous nature of Tn antigens on cancers has made them a natural focus of vaccine research, such carbohydrate moieties are not always tumor-specific and have been observed on embryonic and nonmalignant adult tissue. Here we report the structural basis of binding of a complex of a monoclonal antibody (237mAb) with a truly tumor-specific glycopeptide containing the Tn antigen. In contrast to glycopeptide-specific antibodies in complex with simple peptides, 237mAb does not recognize a conformational epitope induced in the peptide by sugar substitution. Instead, 237mAb uses a pocket coded by germ-line genes to completely envelope the carbohydrate moiety itself while interacting with the peptide moiety in a shallow groove. Thus, 237mAb achieves its striking tumor specificity, with no observed physiological cross-reactivity to the unglycosylated peptide or the free glycan, by a combination of multiple weak but specific interactions to both the peptide and to the glycan portions of the antigen.
- Published
- 2010
- Full Text
- View/download PDF
186. Immunotherapy of lymphoma and leukemia with T-cell engaging BiTE antibody blinatumomab.
- Author
-
Nagorsen D, Bargou R, Ruttinger D, Kufer P, Baeuerle PA, and Zugmaier G
- Subjects
- Antigens, CD19 immunology, B-Lymphocytes cytology, B-Lymphocytes immunology, Cytotoxicity, Immunologic drug effects, Cytotoxicity, Immunologic immunology, Humans, Leukemia immunology, Lymphoma immunology, Models, Immunological, T-Lymphocytes cytology, Antibodies, Bispecific therapeutic use, Immunotherapy methods, Leukemia therapy, Lymphoma therapy, T-Lymphocytes immunology
- Abstract
Hematologic malignancies are predominantly treated with chemotherapy or a combination of chemotherapy with monoclonal antibodies. Current treatment regimens often are not satisfactory due to severe side effects and a substantial proportion of relapsed patients. This has prompted the development of better tolerated and more efficacious immunotherapeutic strategies for the treatment of lymphoma and leukemia. With blinatumomab (MT103) a first antibody, which can potentially engage all cytotoxic T cells of a patient for tumor cell lysis, is now available for clinical evaluation. Blinatumomab is a single-chain bispecific antibody construct with specificity for CD19 and CD3 belonging to the class of bispecific T cell engager. Here, we review the current progress in development of blinatumomab for treatment of patients with CD19-expressing hematological malignancies.
- Published
- 2009
- Full Text
- View/download PDF
187. Antitumor activity of an EpCAM/CD3-bispecific BiTE antibody during long-term treatment of mice in the absence of T-cell anergy and sustained cytokine release.
- Author
-
Amann M, D'Argouges S, Lorenczewski G, Brischwein K, Kischel R, Lutterbuese R, Mangold S, Rau D, Volkland J, Pflanz S, Raum T, Münz M, Kufer P, Schlereth B, Baeuerle PA, and Friedrich M
- Subjects
- Animals, Antibodies, Bispecific adverse effects, Antigens, Neoplasm genetics, Antigens, Neoplasm metabolism, CD3 Complex genetics, CD3 Complex metabolism, Cell Adhesion Molecules genetics, Cell Adhesion Molecules metabolism, Cell Proliferation drug effects, Clonal Anergy drug effects, Cytokines metabolism, Epithelial Cell Adhesion Molecule, Female, Humans, Immunologic Factors administration & dosage, Immunologic Factors adverse effects, Lymphocyte Activation drug effects, Mammary Neoplasms, Experimental pathology, Mammary Neoplasms, Experimental therapy, Mice, Mice, Inbred BALB C, Neoplasm Transplantation, Protein Engineering, T-Lymphocytes drug effects, T-Lymphocytes metabolism, T-Lymphocytes pathology, Time Factors, Antibodies, Bispecific administration & dosage, Antigens, Neoplasm immunology, CD3 Complex immunology, Cell Adhesion Molecules immunology, Immunotherapy, Mammary Neoplasms, Experimental immunology, T-Lymphocytes immunology
- Abstract
muS110 is a BiTE antibody bispecific for murine EpCAM (CD326) and murine CD3. MT110, its human-specific analog, is in a clinical phase 1 trial for treatment of patients with adenocarcinoma of the lung or gastrointestinal tract. Recent studies have shown a therapeutic window for muS110, have explored single-dose toxicity of muS110, and have found that a 1-week low-dose treatment dramatically increased the tolerability of mice to very high doses of muS110 (Cancer Immunol. Immunother. 2009;58:95-109). Here we analyzed the impact of long-term, high-dose treatment of mice with muS110 on antitumor activity and functionality of T cells. After an initial self-limiting cytokine release, the 1-week adaptation period effectively blunted further cytokine production in response to a subsequent high-dose treatment with muS110. The much-increased tolerability of mice adapted to muS110 was not because of anergy of T cells. T cells isolated from chronically muS110-treated mice fully retained their cytotoxic potential, proliferative capacity, and responsiveness to stimulation by either muS110 or anti-CD3/anti-CD28/interleukin-2 when compared with T cells from control mice. Unimpaired T-cell performance was also evident from the effective prevention of orthotopic 4T1 breast tumor outgrowth in mice treated long term with escalating doses of muS110. Finally, we show that muS110 and MT110 recognize orthologous epitopes on mouse and human EpCAM proteins, suggesting that the target-related safety profile of muS110 in mice may be predictive for MT110 in humans.
- Published
- 2009
- Full Text
- View/download PDF
188. Potent control of tumor growth by CEA/CD3-bispecific single-chain antibody constructs that are not competitively inhibited by soluble CEA.
- Author
-
Lutterbuese R, Raum T, Kischel R, Lutterbuese P, Schlereth B, Schaller E, Mangold S, Rau D, Meier P, Kiener PA, Mulgrew K, Oberst MD, Hammond SA, Baeuerle PA, and Kufer P
- Subjects
- Animals, Antibodies, Bispecific immunology, CD8-Positive T-Lymphocytes metabolism, CHO Cells, Carcinoembryonic Antigen blood, Cricetinae, Cricetulus, Humans, Immunotherapy, Mice, Mice, SCID, Recombinant Fusion Proteins immunology, Tetrahydrofolate Dehydrogenase genetics, Tetrahydrofolate Dehydrogenase immunology, Antibodies, Bispecific therapeutic use, CD3 Complex immunology, CD8-Positive T-Lymphocytes immunology, Carcinoembryonic Antigen immunology, Colorectal Neoplasms therapy, Recombinant Fusion Proteins therapeutic use
- Abstract
Carcinoembryonic antigen (CEA, CD66e) is a well-characterized tumor-associated antigen that is frequently overexpressed in tumors. Phospholipases release CEA from tumor cells resulting in high circulating serum levels of soluble CEA (sCEA) that has been validated as marker for progression of colorectal, breast, and lung cancers. sCEA also acts as a competitive inhibitor for anticancer strategies targeting membrane-bound CEA. As a novel therapeutic approach for treatment of tumors expressing CEA on their cell surface, we constructed a series of bispecific single-chain antibodies (bscAb) combining various single-chain variable fragments recognizing human CEA with a deimmunized single-chain variable fragments recognizing human CD3. CEA/CD3-bscAbs redirected human T cells to lyse CEA-expressing tumor cells in vitro and in vivo. Efficient tumor cell lysis was achieved in vitro at bscAb concentrations from 1 pg/mL (19 fM) to 8.9 pg/mL with preactivated CD8 T cells, and 200 to 500 pg/mL with unstimulated peripheral blood mononuclear cell. The cytotoxic activity of a subset of CEA/CD3-bscAbs was not competitively inhibited by sCEA at concentrations that exceeded levels found in the serum of most cancer patients. Treatment with CEA/CD3-bscAbs prevented the growth of human colorectal cancer lines in a severe combined immunodeficiency mouse model modified to show human T cell killing of tumors. A murine surrogate CEA/CD3-bscAb capable of recruiting murine T cells for redirected tumor lysis in immunocompetent mice prevented the growth of lung tumors expressing human CEA. Together, our results reveal a unique opportunity for targeting cytotoxic T cells toward CEA-expressing tumors without being competitively inhibited by sCEA and establish CEA/CD3-bscAb as a promising and potent therapeutic approach.
- Published
- 2009
- Full Text
- View/download PDF
189. Combination of rituximab with blinatumomab (MT103/MEDI-538), a T cell-engaging CD19-/CD3-bispecific antibody, for highly efficient lysis of human B lymphoma cells.
- Author
-
d'Argouges S, Wissing S, Brandl C, Prang N, Lutterbuese R, Kozhich A, Suzich J, Locher M, Kiener P, Kufer P, Hofmeister R, Baeuerle PA, and Bargou RC
- Subjects
- Antibodies, Monoclonal, Murine-Derived, Antigens, CD19 immunology, Antineoplastic Combined Chemotherapy Protocols, CD3 Complex immunology, Caspase 3 metabolism, Caspase 7 metabolism, Drug Synergism, Granzymes, Humans, Lymphoma, B-Cell drug therapy, Lymphoma, B-Cell pathology, Rituximab, Tumor Cells, Cultured, Antibodies, Bispecific pharmacology, Antibodies, Monoclonal pharmacology, Cytotoxicity, Immunologic drug effects
- Abstract
We have compared the cytotoxic activity of rituximab with that of blinatumomab (MT103/MEDI-538), a single-chain CD19-/CD3-bispecific antibody engaging human T cells. Blinatumomab consistently led to a higher degree of lysis of human lymphoma lines than rituximab, and was active at much lower concentration. The cytotoxicity mediated by blinatumomab and rituximab both caused a potent activation of pro-caspases 3 and 7 in target cells, a key event in induction of granzyme-mediated apoptotic cell death. Combination of rituximab with blinatumomab was found to greatly enhance the activity of rituximab, in particular at low effector-to-target cell ratios and at low antibody concentration.
- Published
- 2009
- Full Text
- View/download PDF
190. BiTE: Teaching antibodies to engage T-cells for cancer therapy.
- Author
-
Baeuerle PA, Kufer P, and Bargou R
- Subjects
- Antibodies, Bispecific immunology, Antigens, CD immunology, CTLA-4 Antigen, Humans, T-Lymphocytes, Cytotoxic immunology, Neoplasms immunology, Neoplasms therapy, T-Lymphocytes immunology
- Abstract
Bispecific T-cell-engager (BiTE) antibodies are designed to transiently engage cytotoxic T-cells for lysis of selected target cells. Although this therapeutic concept had been proposed more than two decades ago, BiTE, and also trispecific, antibodies did not achieve clinical proof-of-concept until the past 2 years. Their clinical activity corroborates findings that ex vivo expanded, autologous T-cells derived from tumor tissue, or transfected with specific T-cell receptors, have shown therapeutic potential in the treatment of solid tumors. While these personalized approaches prove that T-cells alone can have considerable therapeutic activity, even in late-stage cancer, they are cumbersome to perform on a broad basis. This is different for cytotoxic T-lymphocyte antigen 4 (CTLA-4) antibodies, which facilitate generation of tumor-specific T-cell clones, and also for bi- and tri-specific antibodies that directly engage a large proportion of patients' T-cells for cancer cell lysis. The potential of global T-cell engagement for human cancer therapy by T-cell-engaging antibodies has yet to be fully investigated.
- Published
- 2009
191. Mode of cytotoxic action of T cell-engaging BiTE antibody MT110.
- Author
-
Haas C, Krinner E, Brischwein K, Hoffmann P, Lutterbüse R, Schlereth B, Kufer P, and Baeuerle PA
- Subjects
- Adenylyl Cyclases metabolism, Amino Acid Chloromethyl Ketones pharmacology, Antigens, Neoplasm immunology, Antigens, Neoplasm metabolism, Apoptosis, CD3 Complex immunology, CD3 Complex metabolism, CD4-Positive T-Lymphocytes drug effects, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes drug effects, CD8-Positive T-Lymphocytes immunology, Caspase Inhibitors, Cell Adhesion Molecules immunology, Cell Adhesion Molecules metabolism, Cell Line, Tumor, Collagen Type XI antagonists & inhibitors, Cytotoxicity, Immunologic immunology, DNA Fragmentation drug effects, Epithelial Cell Adhesion Molecule, Granzymes genetics, Granzymes immunology, Granzymes metabolism, Humans, Lymphocyte Activation drug effects, Neoplasms immunology, Neoplasms pathology, Neoplasms therapy, Single-Chain Antibodies, Antibodies, Monoclonal pharmacology, CD4-Positive T-Lymphocytes metabolism, CD8-Positive T-Lymphocytes metabolism, Cytotoxicity, Immunologic drug effects
- Abstract
MT110 is an EpCAM/CD3-bispecific antibody construct in clinical development for the treatment of patients with adenocarcinoma expressing EpCAM (CD326). Like other members of this antibody class, MT110 can engage resting, polyclonal CD8(+) and CD4(+) T cells for highly potent redirected lysis of target cells. Here we further explored the mechanism of this action. Complete lysis of EpCAM(+) Kato III gastric cancer cells by previously unstimulated T cells was achieved within 48 h. During this period, a high percentage of CD4(+) and CD8(+) T cells became activated and increased expression of granzyme B. This apparently boosted the capacity for serial target cell lysis as studied at very low effector-to-target ratios. Elimination of cancer cells by MT110-redirected T cells involved membrane damage as was evident from nuclear uptake of propidium iodide and release of the cytosolic enzyme adenylate kinase. Redirected T cells also potently triggered programmed cell death in cancer cells as was evident by membrane blebbing, activation of procaspases 3 and 7, fragmentation of nuclear DNA and cleavage of the caspase substrate poly (ADP ribose) polymerase. Chelation of extracellular calcium fully protected cancer cells from lysis by MT110-redirected T cells, while the pan-caspase inhibitor Z-VAD-FMK blocked activation of procaspases, cleavage of poly (ADP ribose) polymerase and fragmentation of nuclear DNA in cancer cells, but could not prevent nuclear uptake of propidium iodide. Soluble factors did not significantly contribute to cancer cell death. Our study shows that MT110 can efficiently gear up the potential of CD8(+) and CD4(+) T cells for serial lysis, and mediate kill of cancer cells predominantly through poreforming and pro-apoptotic components of cytotoxic T cell granules.
- Published
- 2009
- Full Text
- View/download PDF
192. Tumor regression in cancer patients by very low doses of a T cell-engaging antibody.
- Author
-
Bargou R, Leo E, Zugmaier G, Klinger M, Goebeler M, Knop S, Noppeney R, Viardot A, Hess G, Schuler M, Einsele H, Brandl C, Wolf A, Kirchinger P, Klappers P, Schmidt M, Riethmüller G, Reinhardt C, Baeuerle PA, and Kufer P
- Subjects
- Antibodies, Bispecific adverse effects, Antibodies, Bispecific immunology, Antibodies, Bispecific therapeutic use, Antineoplastic Agents adverse effects, Antineoplastic Agents therapeutic use, B-Lymphocytes immunology, Humans, Immunologic Memory, Immunophenotyping, Leukemia, Lymphocytic, Chronic, B-Cell drug therapy, Leukemia, Lymphocytic, Chronic, B-Cell immunology, Lymphocyte Count, Lymphoma, B-Cell immunology, Lymphoma, Follicular immunology, Lymphoma, Mantle-Cell immunology, Recurrence, T-Lymphocytes immunology, Antibodies, Bispecific administration & dosage, Antineoplastic Agents administration & dosage, Lymphoma, B-Cell drug therapy, Lymphoma, Follicular drug therapy, Lymphoma, Mantle-Cell drug therapy, T-Lymphocytes, Cytotoxic immunology
- Abstract
Previous attempts have shown the potential of T cells in immunotherapy of cancer. Here, we report on the clinical activity of a bispecific antibody construct called blinatumomab, which has the potential to engage all cytotoxic T cells in patients for lysis of cancer cells. Doses as low as 0.005 milligrams per square meter per day in non-Hodgkin's lymphoma patients led to an elimination of target cells in blood. Partial and complete tumor regressions were first observed at a dose level of 0.015 milligrams, and all seven patients treated at a dose level of 0.06 milligrams experienced a tumor regression. Blinatumomab also led to clearance of tumor cells from bone marrow and liver. T cell-engaging antibodies appear to have therapeutic potential for the treatment of malignant diseases.
- Published
- 2008
- Full Text
- View/download PDF
193. Therapeutic window of MuS110, a single-chain antibody construct bispecific for murine EpCAM and murine CD3.
- Author
-
Amann M, Brischwein K, Lutterbuese P, Parr L, Petersen L, Lorenczewski G, Krinner E, Bruckmeier S, Lippold S, Kischel R, Lutterbuese R, Kufer P, Baeuerle PA, and Schlereth B
- Subjects
- Animals, Antibodies, Bispecific immunology, Antibodies, Bispecific therapeutic use, Antibodies, Monoclonal immunology, Antibodies, Monoclonal therapeutic use, Antigens, Neoplasm analysis, Breast Neoplasms drug therapy, Breast Neoplasms immunology, CD3 Complex analysis, Cell Adhesion Molecules analysis, Cricetinae, Cytotoxicity, Immunologic, Epithelial Cell Adhesion Molecule, Humans, Lung Neoplasms drug therapy, Lung Neoplasms immunology, Mice, Neoplasms immunology, Single-Chain Antibodies, Tissue Distribution, Antibodies, Bispecific pharmacokinetics, Antibodies, Monoclonal pharmacokinetics, Antigens, Neoplasm immunology, CD3 Complex immunology, Cell Adhesion Molecules immunology, Neoplasms drug therapy
- Abstract
EpCAM (CD326) is one of the most frequently and highly expressed tumor-associated antigens known and recently has also been found on cancer stem cells derived from human breast, colon, prostate, and pancreas tumors. However, like many other tumor-associated antigens used for antibody-based immunotherapeutic approaches, EpCAM is expressed on normal tissues including epithelia of pancreas, colon, lung, bile ducts, and breast. To assess the therapeutic window of an EpCAM/CD3-bispecific single-chain antibody construct of the bispecific T-cell engager (BiTE) class, we constructed murine surrogate of MT110 (muS110) from single-chain antibodies specific for murine EpCAM and CD3 antigens. Immunhistochemical analysis showed that, with minor differences, the expression of EpCAM protein on a large variety of tissues from man and mouse was similar with respect to distribution and level. MuS110 exhibited significant antitumor activity at as low as 5 microg/kg in both syngeneic 4T1 orthotopic breast cancer and CT-26 lung cancer mouse models. Dosing of muS110 for several weeks up to 400 microg/kg by intraanimal dose escalation was still tolerated, indicating existence of a significant therapeutic window for an EpCAM-specific BiTE antibody in mice. MuS110 was found to have similar in vitro characteristics and in vivo antitumor activity as MT110, a human EpCAM/human CD3-bispecific BiTE antibody that currently is in formal preclinical development.
- Published
- 2008
- Full Text
- View/download PDF
194. Strictly target cell-dependent activation of T cells by bispecific single-chain antibody constructs of the BiTE class.
- Author
-
Brischwein K, Parr L, Pflanz S, Volkland J, Lumsden J, Klinger M, Locher M, Hammond SA, Kiener P, Kufer P, Schlereth B, and Baeuerle PA
- Subjects
- Amino Acid Substitution, Animals, Antibodies, Bispecific genetics, Antigens, CD metabolism, Antigens, CD19 metabolism, Antigens, Differentiation, T-Lymphocyte metabolism, Antigens, Neoplasm genetics, CD3 Complex metabolism, CHO Cells, Cell Adhesion Molecules genetics, Cell Proliferation, Cricetinae, Cricetulus, Cytokines metabolism, Epithelial Cell Adhesion Molecule, Epitopes, T-Lymphocyte immunology, Epitopes, T-Lymphocyte metabolism, Humans, Immunoglobulin Fragments genetics, Interleukin-2 Receptor alpha Subunit metabolism, Lectins, C-Type, Leukocytes, Mononuclear immunology, Leukocytes, Mononuclear metabolism, Lymphocyte Activation immunology, Macaca fascicularis, Mice, Muromonab-CD3 metabolism, Recombinant Proteins immunology, T-Lymphocytes metabolism, T-Lymphocytes, Cytotoxic immunology, T-Lymphocytes, Cytotoxic metabolism, Transfection, Antibodies, Bispecific immunology, Antigens, Neoplasm immunology, Cell Adhesion Molecules immunology, Immunoglobulin Fragments immunology, T-Lymphocytes immunology
- Abstract
Bispecific antibodies have been extensively studied in vitro and in vivo for their use in redirected tumor cell lysis. A particular challenge of bispecific antibody constructs that recognize the invariant CD3 signaling complex is a controlled polyclonal activation of T cells that, ideally, is exquisitely dependent on the presence of target cells. Otherwise, overt production of inflammatory cytokines and secondary reactions may occur as side effects, as can be observed with constitutively T-cell activating monoclonal antibodies to CD3 or CD28, and with bispecific antibodies bearing Fc gamma portions. Here we analyzed 2 distinct bispecific single-chain antibody constructs of the BiTE class, called MT110 and MT103 (or MEDI-538), for conditional T-cell activation. In the presence of target-expressing cell lines, low picomolar concentrations of the BiTE molecules were sufficient to stimulate a high percentage of peripheral human T cells to express cytokines and surface activation markers, enter into cell cycle, and induce redirected lysis of target cells. However, in the absence of target cells, the 2 BiTE molecules even at high concentrations did not detectably activate T cells. Our data show that T cell activation by monomeric forms of MT110 and MT103 is highly conditional in that it is strictly dependent on the presence of cells expressing the proper target antigen. BiTE molecules therefore qualify for a highly controlled polyclonal T-cell therapy of cancer.
- Published
- 2007
- Full Text
- View/download PDF
195. A multimarker real-time RT-PCR for MAGE-A gene expression allows sensitive detection and quantification of the minimal systemic tumor load in patients with localized cancer.
- Author
-
Mecklenburg I, Weckermann D, Zippelius A, Schoberth A, Petersen S, Prang N, Riethmüller G, and Kufer P
- Subjects
- Aged, Antigens, Neoplasm genetics, Gene Expression, Humans, Male, Melanoma-Specific Antigens, Middle Aged, Prognosis, RNA, Messenger analysis, RNA, Neoplasm analysis, Sensitivity and Specificity, Transcription, Genetic, Biomarkers, Tumor genetics, Neoplasm Proteins genetics, Prostatic Neoplasms pathology, Reverse Transcriptase Polymerase Chain Reaction methods, Tumor Burden
- Abstract
Introduction: Distant metastases of solid tumors are usually associated with fatal outcome. Disseminated cancer cells are considered early indicators of metastasis. Their sensitive detection and quantification would be a valuable tool for staging of disease and as guidance for therapeutic decisions., Experimental Design: We established a highly sensitive and quantitative multimarker real-time RT-PCR assay for amplification of cancer-related genes MAGE-A1, -A2, -A3/6, -A4, -A10 and -A12 using SYBR green I to detect one single tumor cell in 2 mL of blood or bone marrow. The feasibility of the assay was tested in a large cohort of 177 patients with locally confined prostate carcinoma., Results: Analysis revealed frequent MAGE expression in venous blood and bilateral bone marrow samples (25.5% of all cases) and yielded the first quantitative profile of MAGE expression with a broad range of transcript concentrations for individual markers in the minimal systemic tumor load of patients with localized cancer., Conclusions: Rare transcripts of different MAGE-A genes can be quantified in clinical samples of cancer patients by a sensitive multimarker real-time RT-PCR. Because of frequent expression of MAGE genes in various types of cancer the multimarker MAGE real-time RT-PCR may be generally useful for detection, quantification and characterization of the individual disseminated tumor load in cancer patients.
- Published
- 2007
- Full Text
- View/download PDF
196. Selective targeting and potent control of tumor growth using an EphA2/CD3-Bispecific single-chain antibody construct.
- Author
-
Hammond SA, Lutterbuese R, Roff S, Lutterbuese P, Schlereth B, Bruckheimer E, Kinch MS, Coats S, Baeuerle PA, Kufer P, and Kiener PA
- Subjects
- Animals, Antibodies, Bispecific immunology, Antibody Specificity, CHO Cells, Cell Growth Processes immunology, Cell Line, Tumor, Cricetinae, Cricetulus, Epitopes immunology, Humans, Intercellular Junctions immunology, Lymphocyte Activation, Mice, Mice, Inbred NOD, Mice, SCID, Neoplasms immunology, Neoplasms pathology, T-Lymphocytes immunology, Xenograft Model Antitumor Assays, Antibodies, Bispecific pharmacology, CD3 Complex immunology, Immunization, Passive methods, Neoplasms therapy, Receptor, EphA2 immunology
- Abstract
The EphA2 receptor tyrosine kinase is frequently overexpressed and functionally altered in malignant cells and thus provides opportunities for selective targeting of tumor cells. We describe here the development of a novel, bispecific single-chain antibody (bscAb) referred to as bscEphA2xCD3. This molecule simultaneously targets EphA2 on tumor cells and the T-cell receptor/CD3 complex on T cells and possesses structural and functional characteristics of the recently developed BiTE technology. An EphA2-specific single-chain antibody was selected for recognition of an epitope that is preferentially exposed on malignant cells based on the concept of epitope exclusion; this was fused to a CD3-specific single-chain antibody to generate bscEphA2xCD3. The resultant bscAb redirected unstimulated human T cells to lyse EphA2-expressing tumor cells both in vitro and in vivo. In separate experiments, efficient tumor cell lysis was achieved in vitro at drug concentrations
- Published
- 2007
- Full Text
- View/download PDF
197. CD19-/CD3-bispecific antibody of the BiTE class is far superior to tandem diabody with respect to redirected tumor cell lysis.
- Author
-
Mølhøj M, Crommer S, Brischwein K, Rau D, Sriskandarajah M, Hoffmann P, Kufer P, Hofmeister R, and Baeuerle PA
- Subjects
- Animals, Antibodies, Bispecific genetics, Antibodies, Bispecific pharmacology, Antibodies, Bispecific therapeutic use, Antineoplastic Agents pharmacology, Antineoplastic Agents therapeutic use, Cell Line, Tumor, Dose-Response Relationship, Drug, Humans, Lymphoma drug therapy, Mice, Recombinant Proteins genetics, Recombinant Proteins immunology, Recombinant Proteins pharmacology, Recombinant Proteins therapeutic use, Antibodies, Bispecific immunology, Antibody-Dependent Cell Cytotoxicity, Antigens, CD19 immunology, Antineoplastic Agents immunology, CD3 Complex immunology, Lymphoma immunology
- Abstract
Many kinds of bispecific antibodies recruiting T cells for cancer therapy have been developed. Side-by-side comparison has shown that CD19-/CD3-bispecific antibodies of the diabody, tandem diabody (Tandab) and quadroma format had similar cytotoxic activity, with Tandab being the most active format. Tandab has also been claimed to be superior to single-chain (sc) Fv-based bispecific constructs although data from a side-by-side comparison are not available. In this study, we compared side-by-side MT103 (bscCD19xCD3), a single-chain bispecific antibody of the BiTE class, with a CD19-/CD3-bispecific representative of the Tandab class. Based on literature data, we have constructed, produced and characterized the LL linker version of Tandab, which was reported to be the most active version of Tandab proteins. A dimeric protein of 114kDa was obtained that showed proper bispecific binding to CD3- and CD19-positive cells and could redirect both pre-stimulated and unstimulated human T cells for lysis of human B lymphoma lines Raji, MEC-1 and Nalm-6. Raji cells were lysed at a half-maximal concentration (EC50) of 10 nM Tandab using pre-stimulated T cells, which closely matched the published activity of LL-Tandab with this particular cell line. MT103 had between 700- and 8000-fold higher efficacy than Tandab for redirected lysis of the three human B lymphoma lines. These data demonstrate that under identical experimental conditions, the BiTE format has far superior activity compared to the Tandab format and is also superior to conventional diabody and quadroma formats. The extraordinary potency of the BiTE class and its representative MT103 may translate into improved anti-tumor activity, lower dosing and lower costs of production compared to other bispecific antibody formats.
- Published
- 2007
- Full Text
- View/download PDF
198. Antitumor activity of a dual cytokine/single-chain antibody fusion protein for simultaneous delivery of GM-CSF and IL-2 to Ep-CAM expressing tumor cells.
- Author
-
Schanzer JM, Fichtner I, Baeuerle PA, and Kufer P
- Subjects
- Animals, Antibodies, Neoplasm, Antigens, Neoplasm genetics, Antineoplastic Agents administration & dosage, Cell Adhesion Molecules genetics, Cell Line, Cricetinae, Drug Delivery Systems, Epithelial Cell Adhesion Molecule, Female, Granulocyte-Macrophage Colony-Stimulating Factor administration & dosage, Humans, Immunotherapy, Active, Interleukin-2 administration & dosage, Lung Neoplasms immunology, Lung Neoplasms secondary, Mice, Mice, Inbred BALB C, Neoplasm Transplantation, Recombinant Fusion Proteins administration & dosage, Recombinant Fusion Proteins genetics, Recombinant Proteins, Transplantation, Heterologous, Xenograft Model Antitumor Assays, Antigens, Neoplasm biosynthesis, Antineoplastic Agents therapeutic use, Cell Adhesion Molecules biosynthesis, Granulocyte-Macrophage Colony-Stimulating Factor genetics, Interleukin-2 genetics, Lung Neoplasms drug therapy, Recombinant Fusion Proteins therapeutic use
- Abstract
Cytokine targeting to tumor-associated antigens via antibody cytokine fusion proteins has demonstrated potent antitumor activity in numerous animal models and has led to the clinical development of 2 antibody-interleukin-2 (IL-2) fusion proteins. We previously reported on the construction and in vitro properties of a "dual" cytokine fusion protein for simultaneous targeted delivery of human granulocyte macrophage-colony stimulating factor (GM-CSF) and IL-2 to human tumors. The fusion protein is based on a heterodimerized core structure formed by human CH1 and Ckappa domains (heterominibody) with C-terminally fused human cytokines and N-terminally fused single-chain antibody fragments specific for the tumor-associated surface antigen epithelial cell adhesion molecule (Ep-CAM). For testing the antitumor activity in syngeneic mouse xenograft models, we developed "dual cytokine heterominibodies" with murine cytokines (mDCH). mDCH fusion proteins and, as controls, "single cytokine heterominibodies" (SCH) carrying either murine GM-CSF (mGM-CSF) or murine IL-2 (mIL-2) were constructed, of which all retained the specific activities of cytokines and binding to the Ep-CAM antigen on human Ep-CAM transfected mouse colon carcinoma CT26-KSA cells. Over a 5-day treatment course, DCH fusion proteins induced significant inhibition of established pulmonary CT26-KSA metastases in immune-competent Balb/c mice at low daily doses of 1 mug of fusion protein per mouse. However, with the tested dosing schemes, antitumor activity of mDCH was largely independent of cytokine targeting to tumors as demonstrated by a control protein with mutated Ep-CAM binding sites. Single cytokine fusion proteins mSCH-GM-CSF and mSCH-IL-2 showed similar antitumor activity as the dual cytokine fusion protein mDCH, indicating that GM-CSF and IL-2 in one molecule did not significantly synergize in tumor rejection under our experimental conditions. Our results seem to contradict the notion that IL-2 and GM-CSF can synergize in antitumor activity and that with conventional dose regimens, their specific targeting to tumors, as tested here with 2 antibodies of different affinities, enhances their antitumor activity.
- Published
- 2006
- Full Text
- View/download PDF
199. Potent inhibition of local and disseminated tumor growth in immunocompetent mouse models by a bispecific antibody construct specific for Murine CD3.
- Author
-
Schlereth B, Kleindienst P, Fichtner I, Lorenczewski G, Brischwein K, Lippold S, da Silva A, Locher M, Kischel R, Lutterbüse R, Kufer P, and Baeuerle PA
- Subjects
- Animals, Antibodies, Bispecific immunology, Antibodies, Bispecific pharmacokinetics, Antibody Specificity, Cell Line, Tumor transplantation, Dose-Response Relationship, Immunologic, Drug Screening Assays, Antitumor, Epithelial Cell Adhesion Molecule, Humans, Immunocompetence, Lung Neoplasms secondary, Lung Neoplasms therapy, Melanoma, Experimental immunology, Mice, Mice, Inbred C57BL, Mice, Inbred NOD, Mice, SCID, Neoplasm Transplantation, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma pathology, Recombinant Fusion Proteins immunology, Subcutaneous Tissue, Xenograft Model Antitumor Assays, Antibodies, Bispecific therapeutic use, Antigens, Neoplasm immunology, CD3 Complex immunology, Cell Adhesion Molecules immunology, Immunotherapy, Melanoma, Experimental therapy, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma therapy
- Abstract
Bispecific single-chain antibody constructs specific for human CD3 have been extensively studied for antitumor activity in human xenograft models using severe combined immunodeficient mice supplemented with human T cells. High efficacy at low effector-to-target ratios, independence of T cell costimuli and a potent activation of previously unstimulated polyclonal T cells were identified as hallmarks of this class of bispecific antibodies. Here we studied a bispecific single-chain antibody construct (referred to as 'bispecific T cell engager', BiTE) in an immunocompetent mouse model. This was possible by the use of a murine CD3-specific BiTE, and a syngeneic melanoma cell line (B16F10) expressing the human Ep-CAM target. The murine CD3-specific BiTE, called 2C11x4-7 prevented in a dose-dependent fashion the outgrowth of subcutaneously growing B16/Ep-CAM tumors with daily i.v. injections of 5 or 50 microg BiTE which was most effective. Treatment with 2C11x4-7 was effective even when it was started 10 days after tumor cell inoculation but delayed treatments showed a reduction in the number of cured animals. 2C11x4-7 was also highly active in a lung tumor colony model. When treatment was started on the day of intravenous tumor cell injection, seven out of eight animals stayed free of lung tumors, and three out of eight animals when treatment was started on day 5. Our study shows that BiTEs also have a high antitumor activity in immunocompetent mice and that there is no obvious need for costimulation of T cells by secondary agents.
- Published
- 2006
- Full Text
- View/download PDF
200. T-cell activation and B-cell depletion in chimpanzees treated with a bispecific anti-CD19/anti-CD3 single-chain antibody construct.
- Author
-
Schlereth B, Quadt C, Dreier T, Kufer P, Lorenczewski G, Prang N, Brandl C, Lippold S, Cobb K, Brasky K, Leo E, Bargou R, Murthy K, and Baeuerle PA
- Subjects
- Animals, Antigens, CD19 immunology, B-Lymphocytes immunology, CD3 Complex immunology, Cell Line, Tumor, Cells, Cultured, Cytotoxicity Tests, Immunologic, Female, Flow Cytometry, Humans, Immunotherapy, Leukocytes, Mononuclear drug effects, Leukocytes, Mononuclear immunology, Lymphocyte Activation immunology, Male, Pan troglodytes, T-Lymphocytes immunology, Antibodies, Bispecific pharmacokinetics, B-Lymphocytes drug effects, Lymphocyte Activation drug effects, Lymphocyte Depletion, T-Lymphocytes drug effects
- Abstract
BscCD19xCD3 is a bispecific single-chain antibody construct with exceptional cytotoxic potency in vitro and in vivo. Here, we have investigated the biological activity of bscCD19xCD3 in chimpanzee, the only animal species identified in which bscCD19xCD3 showed bispecific binding, redirected B-cell lysis and cytokine production comparable to human cells. Pharmacokinetic analysis following 2-h intravenous infusion of 0.06, 0.1 or 0.12 mug/kg of bscCD19xCD3 as part of a dose escalation study in a single female chimpanzee revealed a half-life of approximately 2 h and elimination of the bispecific antibody from circulation within approximately 8 h after the end of infusion. This short exposure to bscCD19xCD3 elicited a transient increase in serum levels of IFNgamma, IL-6, IL-2, soluble CD25, and transiently upregulated expression of CD69 and MHC class II on CD8-positive cells. Cytokine release and upregulation of T-cell activation markers were not observed with vehicle controls. A multiple-dose study using 5 weekly doses of 0.1 mug/kg in two animals also showed transient cytokine release and an activation of peripheral T cells with a first-dose effect, accompanied by a transient lymphopenia. While oscillations of T-cell counts were relatively even during repeated treatments, the amplitudes of peripheral B cells declined with every infusion, which was not observed in a vehicle control animal. Our data show that bscCD19xCD3 can be safely administered to chimpanzees at dose levels that cause fully reversible T-cell activation and, despite a very short exposure time, cumulative loss of peripheral B lymphocytes. A clinical trial testing prolonged administration of bscCD19xCD3 (MT103) for improving efficacy is currently ongoing.
- Published
- 2006
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.