bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bleomycin Pulmonary Toxicity Does Impact on Hodgkin’s Disease Outcome.
Authors: Martin William G. , Ristow Kay M. , Colgan Joseph P. , Habermann Thomas M. , Ansell Stephen M. 
Published: April 2020

Abstract
Background: Bleomycin-containing chemotherapy regimens have become the mainstay of treatment in Hodgkin’s Disease (HD). The risk of bleomycin pulmonary toxicity (BPT) ranges from 0–46% and mortality rates range from 0–27%. Although there is no excepted standard treatment for BPT, corticosteroid treatment, withholding bleomycin from subsequent chemotherapy, and proceeding with a non-bleomycin containing regimen is the most common approach. We reviewed our experience to better delineate outcome and appropriate treatment in these patients.
Methods: We reviewed all patients managed for HD at Mayo Clinic, Rochester, from January 1986 to February 2003. BPT was defined by the presence of pulmonary symptoms, bilateral interstitial infiltrates, and no evidence for an infectious etiology.
Results: A retrospective review identified 141 patients with HD treated with a bleomycin-containing regimen. The mean age of patients at diagnosis was 34 years. Fifty four percent were males. The histology was nodular sclerosing in 85%. The Hasenclever index was 0 in 15%, 1 in 35%, 2 in 24%, 3 in 16%, 4 in 9%, and &gt;5 in 1%. Frontline chemotherapy included ABVD in 57%, MOPP-ABV(D) in 33%, COPP-ABV in 8%, BEACOPP in 3%, and Stanford V in 2%. Forty one percent of patients received radiation. The 5-year overall survival (OS) and progression free survival (PFS) for all patients were 87% and 79% respectively. BPT was observed in 18% of patients (25/141). There was a significant decrease in OS, 63% in patients with BPT versus 90% in non-BPT patients at 5 years (p=0.0013). The mortality from BPT was 4.2% (6/141) in all patients and 24% (6/25) in patients who developed the pulmonary syndrome. These 6 patients all died within 9 months of their HD diagnosis from BPT. The Hasenclever index was 3 or less in 5 of these 6 patients. CR rates were equal at 91% and 93% in BPT and non-BPT patients respectively (p=0.299). Twenty two percent (31/141) of patients, either symptomatic or asymptomatic, had bleomycin omitted at any time from their regimen with no difference in OS or PFS, 83% versus 88% (p=0.369) and 82% versus 78% (p=0.853) respectively. The mean bleomycin dose was 84 mg/m2 (range of 20–180 mg/m2). Bleomycin dose had no impact on OS or PFS. In BPT patients subsequent non-bleomycin therapy included AVD 44%, MOPP-AV 31%, or no further chemotherapy 25%. The five-year OS was equal between the AVD and MOPP-AV groups at 91%. However in patients who received no further chemotherapy 5-year OS was 30% (p=0.0001).
Conclusion: Bleomycin pulmonary toxicity (BPT) results in a significant decrease in OS survival at 5 years in patients being treated for Hodgkin’s Disease. In patients who do not die acutely from pulmonary toxicity both OS and PFS appear equal despite the omission of bleomycin.
Preliminary Results of a Phase I/II Trial of BBR-2778 (Pixantrone) in Combination with Fludarabine, Dexamethasone, and Rituximab (FPD-R) in the Treatment of Patients with Relapsed/Refractory Indolent Non-Hodgkin’s Lymphoma (NHL).
Authors: Fayad Luis , Liebmann James , Modiano Manuel , Cohen Gary I. , Pro Barbara , Romaguera Jorge , Davite Cristina , Lymboura Margarita , Comis Silvia , Laffranchi Bernard L. 
Published: April 2020

Abstract
Background: Fludarabine-mitoxantrone-dexamethasone-rituximab (FND-R) is a combination chemotherapy with significant activity in untreated and relapsed indolent B-cell lymphoma patients (pts). Pixantrone is a novel aza-anthracenedione that has no delayed cardiotoxicity in animal models and has single agent activity in NHL. We conducted an outpatient phase I/II trial, to define the recommended dose (RD) of pixantrone when substituting for mitoxantrone in the FND-R regimen, and the safety and efficacy of this regimen in pts with relapsed or refractory indolent NHL.
Methods: Pts received pixantrone with fludarabine (25mg/m2/day, days 1–3), dexamethasone (20mg/day, days 1–5), and rituximab (375mg/m2/day on day 1) in a 28 day regimen (FPD-R). Based on previous single agent clinical studies, the starting dose of pixantrone was defined as 80mg/m². The RD was established to be 120mg/m2 and the protocol was amended to treat patients in the phase II part.
Results: 9 pts (6 males) with a median age of 65 years (range 41–78) were included in the dose-finding part of the study. Following the protocol amendment, 23 pts (11 males) of WHO performance status 0–1, median age 61 (range 32–78) have been enrolled in the currently ongoing phase II part. Pathology included SLL/CLL, follicular grade I, follicular grade II, MALT, marginal cell, and other indolent B-cell lymphomas. All pts had received a prior anthracycline-containing regimen (median number 1, range 1–4). The study regimen was well tolerated; median number of courses received was 5 (range 2–8). Grade 4 toxicities were neutropenia in 10 pts, leukopenia in 5 pts, and thrombocytopenia in 1 pt. No clinically significant cardiac events or decreases in LVEF ≥20% were noted. However 4 pts went below 50% LVEF including 1 pt who was below 50% at baseline. Non-hematologic adverse events were primarily grade 1 or 2 in severity. Response rate was 75% for the 20 pts evaluable for response, with 11 (55%) complete remission [7 confirmed (CR), 4 unconfirmed (CRu)], and 4 (20%) partial remission (PR). Two responders went onto bone marrow transplant (BMT): one had a CRu and one a PR. A third patient was pending BMT after CR. At a median follow-up of 345 days, the median duration of response has not been reached, as only two pts have progressed: the first progression occurred at 113 days, and the second pt progressed at 699 days. The remaining pts are still in remission.
Conclusions: The RD of pixantrone in the FPD-R regimen is 120mg/m2. The primary toxicity is hematologic. The regimen can be given on an outpatient basis, is associated with major responses, and is very well tolerated in relapsed and refractory indolent NHL pts.
Immunodeficiency Associated with a STAT-5b Mutation and Growth Hormone Insensitivity.
Authors: Cleary Aileen M. , Hwa Vivian , Tu Wenwei , Dionis Kira , Bezrodnik Liliana , Teper Alejandro , Gaillard Maria , Heinrich Juan J. , Rosenfeld Ron G. , Lewis David B. 
Published: April 2020

Abstract
We recently identified a homozygous mutation in the STAT-5b (signaling transducer and activator of transcription 5b) gene in a female patient with growth hormone insensitivity. The patient had significant growth retardation (height -7.5 SD), despite normal levels of growth hormone (GH) and GH binding protein. Analysis of the GH receptor signaling pathways indicated that the mutation (Ala630 to Pro630) resulted in a protein that could not be activated by GH, and subsequent dysregulation of STAT-5b-dependent genes, such as IGF-I and IGFBP-3. This abstract details an immunodeficiency due to a lack of functional STAT5b. Clinically, the patient had chronic diarrhea as an infant, and suffered several severe infections, including persistent lymphocytic interstitial pneumonia (LIP), a condition associated with children who are HIV-positive or immunodeficient. Laboratory studies showed hypergammaglobulinemia, T-cell lymphopenia with a normal CD4:CD8 T-cell ratio, and total B and NK cells within the normal range. Immunologic analysis showed decreased interleukin (IL)-2 receptor α expression in response to IL-2 on both CD4 and CD8 T cells, despite normal proliferation to mitogens, and normal anti-CD3 proliferation by CFSE analysis. IL-2 mediated γc (common gamma chain) upregulation, induction of CD40-ligand and interferon-γ production by intracellular cytokine analysis in response to either the bacterial superantigen SEB or anti-CD3/anti-CD28 mAb stimulation were normal, suggesting that STAT5b is not necessary for these functions. Importantly, perforin expression in response to IL-2 was decreased in CD8 T cells suggesting that both decreased CD8 number and function contribute to this immunodeficiency. Interestingly, NK cells failed to increase adhesion molecules CD54 or CD58 in response to IL-2 suggesting impaired NK cell function. Lastly, EBV transformed B cell lines were unable to phosphorylate STAT5b in response to IL-2 and IL-7 suggesting an intrinsic B cell defect that may affect immunoglobulin production. Thus, the STAT5b −/ − mutation appears to limit the effects of important γc-containing cytokines on CD4, CD8, NK and B cells, resulting in increased susceptibility to infection.
Tumor Localization and Quantitation of Adoptively Transfered T Lymphocytes in a Murine Model.
Authors: Hokland Marianne , Petersen Mikkel S. , Fleischer Charlotte C. , Stødkilde-Jørgensen Hans , Hansen Søren B. , Skands Ulrik , Gundersen Hans J.G. , Agger Ralf 
Published: April 2020

Abstract
Tracking adoptively transferred antigen-specific T lymphocytes is an important prerequisite for devising better protocols for cellular therapy. To this end we have developed a highly sensitive method for “in situ” visualization of labelled lymphocytes in vivo by combined PET and magnetic resonance imaging (MRI) to monitor the distribution of adoptively transferred tumour-specific T cells in a mouse model system. Moreover, quantitation of the adoptively transferred cells in tumor was performed by flow cytometry.
C57BL/6J mice carrying subcutaneous tumours of the ovalbumin (OVA)-expressing malignant melanoma cell line B16-OVA were adoptively transferred with OVA-specific CD8+ T cells labelled with 124IdU. Five days after transfer of T cells, mice were killed and subjected to PET and MR imaging. Using a newly developed method for co-registration of the two image modalities, the anatomical localisation of the transferred cells was visualised and the amount of radioactivity in various anatomical locations very accurately determined.
For quantitation of tumor infiltrating non-labelled OVA-specific CD8+ T cells by flow cytometry (using AbsoluteCount Beads), tumors were removed from mice day 1 until day 8 following adoptive transfer (6 mice/group) and prepared for single cell suspension before labeled with anti-CD8-FITC and SIINFEKL-Tetramer-PE.
Results showed a clear tumor localization of the adoptively transferred OVA-specific T cells in the tumours. In two independent experiments comprising 12 and 13 evaluable mice, respectively, we found a mean value of 0.909 +/− 0.468 Bq and 0.926 +/− 0.553 Bq in the tumours, and only 0.182 +/− 0.479 Bq and 0.026 +/− 0.480 Bq in the corresponding contralateral control volumes. The difference in activity between the tumour regions and the control regions was statistically highly significant with 2p-values of 0.002 and 0.006 for the two experiments. Using flow cytometry it was shown that the number of OVA specific T lymphocytes accumulating in tumor gradually increased until day 5 after transfer when an average of 3.3 million SIINFEKL-specific cells per gram tumor tissue was found. From day 5 until day 8 the number of SIINFEKL-specific cells per gram tumor tissue fluctuated at a fairly constant level.
This method presented for tracking adoptively transfered tumor specific T lymphocytes represent a significant advancement for studies of adoptively transferred specific T cells, and could potentially be developed for diagnostic purposes. Moreover, since these studies show that tumor-specific T cells home to subcutaneous tumours in substantial numbers, we suggest that these migrating cells could be employed in a new form of therapy as carriers of toxic substances to tumors.
Efficient and Reproducible Retroviral Infection of Primary Human Natural Killer Cells.
Authors: Becknell Brian , Trotta Rossana , Yu Jianhua , Ding Wei , Mao Hsiaoyin C. , Hughes Tiffany , Marburger Trent , Caligiuri Michael A. 
Published: April 2020

Abstract
Molecular characterization of human natural killer (NK) cells will require targeted gene delivery to inhibit and activate specific signaling pathways, yet to our knowledge, an effective means to deliver such products for long-term gene expression without disrupting normal cellular processes has not been described. In this study we have developed a retroviral strategy to effectively express gene products in the NK cell, whereby its effector functions of cytotoxicity and cytokine production remain intact. Using an EBV/retroviral hybrid vector PINCO, we demonstrate infection of human peripheral blood NK cells with simultaneous expression of a marker for infection - the enhanced green fluorescent protein (EGFP) - along with various genes of interest. This technique results in successful infection of the CD56dim NK population that predominates among human peripheral blood NK and is the effector of antibody-dependent cellular cytotoxicity (ADCC) and natural killing. In addition, we demonstrate infection of the CD56bright NK subset as well as the NK-92 and NK-L cell lines. Finally, we modify PINCO to express a cytoplasmically truncated murine CD8 molecule in place of GFP. The resulting vector enables us to transduce NK cells with multiple genes of interest simultaneously and provides an alternative purification method to FACS by using magnetic beads. In summary, we have devised an efficient and highly reproducible methodology for the targeted delivery of gene products to human NK cells that should now provide opportunities to dissect the molecular processes critical to normal NK cell physiology and to genetically manipulate NK cell populations prior to their administration in cancer therapy.
In Vitro Induction of Myeloid Leukemia Specific CD4 and CD8 T-Cells by CD40 Ligand Activated B Cells Gene-Modified to Express Primary Granule Proteins.
Authors: Fujiwara Hiroshi , Melenhorst Joseph J. , El Ouriaghli Frank , Kajigaya Sachiko , Grube Matthias , Sconocchia Giuseppe , Rezvani Katayoun , Price David A. , Hensel Nancy F. , Douek Daniel C. , Barrett A. John 
Published: April 2020

Abstract
The closely-related primary granule proteins are a source of multiple antigens with immunotherapeutic potential for myeloid leukemias. To further explore the possibility of generating leukemia-specific T cell responses, DNA vectors encoding the primary granule proteins proteinase 3 (PR3), human neutrophil elastase (HNE) and cathepsin-G (CathG) were transfected into autologous CD40-activated B (CD40-B) cells and used to stimulate T cell responses from peripheral blood T cells of five patients with myeloid malignancies and three ALL patients three months post-bone marrow transplantation (BMT), and one patient with CML-BC pre-transplant. T-cell responses were measured by flow-based intra-cytoplasmic IFN-g assay and cytotoxic assay. CD8 and CD4 cell response to PR3 and HNE were observed in CD8+ and CD4+ T-cells in 6/6 patients with myeloid leukemias with various degrees. T-cell response against CathG were found in both myeloid and lymphoid leukemias. We conclude that gene-transfected CD40-activated B cells are fully capable of expanding functionally-competent primary granule protein-specific CD4 and CD8 T cells, making this approach a valuable means of expanding leukemia antigen-specific T cells for adoptive immunotherapy.
PGP-DNA primed T cells showed CTL activity to CML cell (case#5, post transplant)
CTL activity % ( E:T ratio ) CML cell; recipient BM cell, normal BM; HLA identical DNR BM cells Effector cells Target 50:1 25:1 12.5:1 PR3-primed CML cell 63.8 54.6 30.4 normal BM 15.5 13.1 5.9 HNE-primed CML cell 43.9 38.3 32.1 normal BM 0 4.1 2.3 CathG-primed CML cell 43.1 29.1 27.5 normal BM 0 0 1.7 mock-primed CML cell 33.3 34.2 29.5 normal BM 5.5 12.9 0
Cotransfection of Dendritic Cells with RNA Coding for a Tumor Antigen and Costimulatory Molecule 4-1BBL Increases the Induction of Her-2/neu Specific Cytotoxic T-Cells.
Authors: Grünebach Frank , Kayser Katrin , Appel Silke , Weck Markus M. , Müller Martin R. , Reichardt Volker L. , Brossart Peter 
Published: April 2020

Abstract
Dendritic cells are professional antigen-presenting cells (APC) with the unique capacity to initiate primary immune responses. Recently, it was demonstrated that transfection of DC with RNA coding for specific antigens can elicit an effective T cell response in vitro and in vivo.
4-1BB (CD137) is a member of a family of receptors found on the surface of cells of the immune system. It plays an important role in the costimulation of T-cells: it enhances T-cell proliferation, cytokine production and cytotoxic effector functions and suppresses the induction of apoptosis. It is predominantly expressed on activated T-cells, and its ligand 4-1BBL is expressed on activated APC including DC.
In the present study we analyzed the effects of 4-1BBL on the stimulatory capacity of DC. Monocyte derived DC were electroporated with in vitro transcribed RNA encoding the tumor-associated antigen Her-2/neu. Additionally these cells were cotransfected with in vitro transcribed RNA coding for the human 4-1BBL with the aim to enhance the costimulatory functions of the cells and therefore the induction of Her-2/neu specific cytotoxic T-cells (CTL).
Standard 51Cr-release assays performed after two restimulations showed specific cytotoxic activity of the induced CTL against autologous DC transfected with Her-2/neu RNA and against the HLA-matched allogeneic Her-2/neu+/HLA-A2+ renal cell carcinoma cell line A498 whereas DC transfected with irrelevant RNA as well as Her-2/neu-/HLA-A2+ CROFT cells and the Her-2/neu+/HLA-A2- cell line SK-OV-3 were spared. There was no lysis of K562 cells, thus excluding a NK-cell mediated killing. In this experimental setting the specific lysis rate of target cells could nearly be doubled by the coadministration of Her-2/neu RNA and an equal amount of 4-1BBL RNA (50% vs 90% specific lysis at effector to target ratio of 30:1). Interestingly, overexpression of 4-1BBL had no effect on the stimulation of allogeneic T cells by electroporated DC in a mixed lymphocyte reaction, indicating that 4-1BBL increases the induction of antigen specific T cell responses rather than the unspecific stimulation of bulk T lymphocytes.
Furthermore, FACS analysis of DC transfected with 4-1BBL plasmid DNA or in vitro transcribed 4-1BBL RNA showed higher expression of the costimulatory molecules CD80, CD86, CD40 and the T-cell adhesion molecule CD54 as compared to the controls indicating that 4-1BBL signaling can mediate activation of DC and thus increase their stimulatory capacity. In the context of immunotherapies for cancer patients the findings of our study could contribute to optimise the in vitro manipulation of DC for the induction of antigen-specific CTL responses.
Transduction of Tolerogenic Dendritic Cells by Foamy Virus and Lentivirus Vectors.
Authors: Su RuiJun , Josephson Neil C. 
Published: April 2020

Abstract
The induction and maintenance of antigen specific peripheral tolerance is critical for the prevention of autoimmunity and maintenance of immune homeostasis. Antigen presentation by tolerogenic dendritic cells (DCs) initiates the process through the stimulation of regulatory T cells and other mechanisms that may include peripheral deletion and anergy induction of autoreactive T cells. In contrast to DCs that mediate immune priming, tolerogenic DCs have a resting phenotype. They lack expression of co-stimulatory molecules and MHC class II. Recently, a method for identifying murine tolerogenic DCs (lin−, CD45RBhigh, CD11clow) and expanding them ex vivo has been described (Wakkach et al. Immunity18:605, 2003). Directed antigen presentation by viral vector transduction of these cells has the potential to produce antigen specific tolerance for therapeutic purposes. To generate tolerogenic DCs, we cultured lin− murine bone marrow in complete RPMI medium in the presence of 10 ng/ml GM-CSF, 2.5 ng/ml TNF-α and 10–100 ng/ml IL-10 for 6–7 days. Cells were transduced overnight at different time points (day 0, day 3, day 6), with foamy virus vectors and lentivirus vectors containing either the GFP or alkaline phosphatase reporter genes. Transductions were performed both in the presence or absence of fibronectin fragment CH-296. For day 0 transductions, we also evaluated whether stem cell stimulatory cytokines (50 ng/ml TPO, 100 ng/ml SCF, 100 ng/ml FL) improved gene transfer efficiency. On days 6–7 the CD45RBhighCD11clow cell population was flow sorted and then further analyzed. The percentage of CD45RBhighCD11clow cells harvested on days 6–7 varied from 10% to 30% and was increased by IL-10 in a dosage dependent manner. As expected for tolerogenic DCs, these cells were morphologically immature by Giemsa-Wright staining and phenotypically negative to low for the expression of co-stimulatory molecules CD80 and CD86, and MHC class II, even after exposure to lipopolysaccharide. At an MOI of 0.5–1, transduction rates of CD45RbhighCD11clow cells were 30%–37% for foamy virus vectors and 23%–24% for lentivirus vectors. Most importantly, vector transduction did not induce expression of co-stimulatory molecules, suggesting that these two viral vectors do not activate DCs. There was no difference in transduction efficiency at the three different time points tested. Furthermore, transduction efficiencies were not affected by either stem cell stimulatory cytokines or CH-296. In summary, cells with the phenotype of tolerogenic DCs can be efficiently transduced by foamy virus and lentivirus vectors with overnight exposure to vector stocks. In vivo studies in mice are in progress to evaluate if transduced CD45RBhighCD11clow cells are able to induce antigen specific tolerance. These studies have potential clinical implications for the treatment of autoimmune diseases, the induction of tolerance after stem cell transplantation, or the prevention of immune responses triggered by specific transgenes in gene therapy.