bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mechanism for the Immuno-Suppresion Activity of Pentostatin: Selective Apoptosis of Naive T Cell Subsets.
Authors: Smith Melody M. , Powers William , Giver Cindy R. , Waller Ned , Flowers Christopher 
Published: October 2019

Abstract
Background: Nucleoside analogs have potent anti-tumor activity, especially against lymphoid malignancies, with immuno-suppression being a significant toxicity. We have previously described the immunosuppressive effects of fludarabine on murine T-cells, and found selective cytotoxicity against naïve subsets that reduced their allo-reactivity in a GvHD model of murine BMT (Giver et al 2003 BBMT 9:616). In this study we tested the cytotoxicity of fludarabine and pentostatin against human and murine T-cell subsets to determine their immune-suppressive activity, mechanism of action, and their potential utility as agents to decrease the allo-reactivity of allogeneic donor lymphocyte infusions.
Methods: Peripheral blood mononuclear cell samples from normal donors were incubated ex vivo ex vivo treatment with Flu at doses of 5, 50, and 250 mcg/ml or pentostatin at doses of 1, 10, and 100 mcg/ml at 0, 4, 24, and 48-hour time points following 1 or 24 hour drug exposure. All samples that obtained dCF also received 25 mcg/ml of deoxyadenosine (dAdo), which enables dCF to mediate cytotoxicity against lymphocytes. Control samples for fludarabine and pentostatin treatment were untreated or treated with dAdo only, respectively. The total number of viable cells and the fractions of the CD4 and CD8 memory and naive T cell subsets that survived nucleoside exposure were determined by multi-parameter flow cytometry following staining with antibodies to CD3, CD4, CD8, CD45RA, CD6L, Annexin, and 7-AAD.
Results: The decrease in the viability of the human naïve CD4 T cells, whether treated with fludarabine or pentostatin, was rapid and most notably observed after the 24-hour time point. On the other hand, the human memory CD4 T cells displayed a more gradual decline in percent viability in both treatment with fludarabine and pentostatin. Furthermore, the potency of pentostatin was exhibited in that it achieved a comparable effect to fludarabine while at substantially lower doses. Data obtained from the murine model also demonstrated a relative a relative sensitivity of the CD4 naïve T cells as oppesed to the CD4 memory to purine analog exposure.
Conclusions: Future work will determine the optimal dosage and period of incubation for pentostatin and fludarabine that can utilized to induce immunosuppresion ex vivo in donor lymphocytes.
SELECTIVE DEPLETION OF NAÏVE CD4+ T-CELLS FOLLOWING IN VITRO EXPOSURE TO PURINE ANALOGS SELECTIVE DEPLETION OF NAÏVE CD4+ T-CELLS FOLLOWING IN VITRO EXPOSURE TO PURINE ANALOGS
Pre-Transplant Biomarkers Predict GVHD and Treatment-Related Mortality in Patients Treated with Extracorporeal Photopheresis Prior to Allogeneic Bone Marrow Transplantation.
Authors: O’Brien Peter J. , Bolwell Brian , Abhyankar Sunil , Shaughnessy Paul , van Besien Koen , Williamson Paul , Campbell Kim A. 
Published: October 2019

Abstract
Accurate prediction of the onset and severity of GvHD and treatment-related mortality (TRM) can lead to improved outcomes through optimized donor selection and GvHD prophylaxis. Current assessments are typically made after bone marrow transplantation (BMT), and often require inherently unreliable qualitative measures. The persistence of host antigen presenting cells (APC), particularly dendritic cells (DC) has been implicated in the onset of GvHD, and a recent clinical trial sought to confirm previous reports that extracorporeal photopheresis (ECP) can prevent GvHD after allogeneic BMT. We examined blood from a subset of 32 ECP-treated patients to identify peripheral blood cell surface markers that might predict GvHD and TRM. Samples were drawn immediately prior to ECP (at baseline) and immediately after ECP, but prior to myeloablative conditioning, then assayed by flow cytometry. Cytometry data were grouped and modeled to assess their predictive accuracy alone or in combination with clinical laboratory values. Logistic regression showed that markers for specific DC subsets present prior to myeloablation were the best predictors of outcomes. The likelihood of grade II-IV GvHD (aGvHD) increases when baseline lin− HLA-DR+ CD11c+ myeloid cells make up a smaller proportion of circulating mononuclear cells with a predictive accuracy, reflected in the area under the receiver-operator curve (ROC) of 0.83. The best predictive model for TRM was a lower absolute abundance of circulating lin− HLA-DR+ CD123+ plasmacytoid DC at baseline (ROC=0.86). No additional predictive power arose with respect to aGvHD or TRM after including laboratory values. However, models that combined certain clinical laboratory results and demographic factors also predicted these clinical outcomes, and offer a possible alternative to complex cytometric assays. The best such model included baseline measurements of BUN/Creatinine ratio, serum albumin, and the match/relatedness of the graft donor and recipient (ROC=0.82, n=59) for TRM, while baseline neutrophil counts were most predictive of aGvHD (ROC=0.69, n=60). In summary, we have identified biomarkers including host DC subsets, readily measured clinical tests, and demographic factors that, at least in patients receiving ECP, are present before conditioning and can predict outcomes. Although this study does not address the impact of ECP on antigen presenting cells directly, the DC profiles described here highlight the potential role of ECP in modulating DC, and thereby decreasing the risks associated with BMT. These results encourage larger studies with non-ECP control populations to directly address this issue.
Multiple Myeloma Patients Treated with Allogeneic Stem Cell Transplantation Show an Increased Frequency of Bone Marrow-Residing CD4+Foxp3+ T Regulatory Cells.
Authors: Atanackovic Djordje , Cao Yanran , Faltz Christiane , Bartels Katrin , Wolschke Christine , Brummendorf Tim H. , Zander Axel R. , Bokemeyer Carsten , Kroeger Nicolaus 
Published: October 2019

Abstract
BACKGROUND: Immunosuppressive CD4+Foxp3+ T regulatory cells (Treg) play a vital role in immune regulation. Thus, Treg contribute to the prevention of autoimmune disease and graft-versus-host reactions following allogeneic stem cell transplantation (alloSCT) but also to the inhibition of effective anti-tumor T cell responses. It has previously been suggested that the frequency of Treg is increased in the peripheral blood of patients with multiple myeloma (MM). However, little is known about the presence of Treg in the bone marrow and it is unclear whether allogeneic stem cell transplantation might deplete Treg from this immune compartment.
METHODS: In the present study, we analyzed percentages of CD4+Foxp3+ Treg as well as Treg expression of CD45RA and CCR7 in the bone marrow (BM) and in the peripheral blood of MM patients who had received alloSCT (N=42), in newly diagnosed MM patients (N=18), and in healthy controls (N=15) using flow cytometry. In addition, we performed inhibition assays in order to test the functional relevance of peripheral and BM-residing Treg.
RESULTS: While newly diagnosed MM patients and healthy controls showed no significant difference in the proportions of CD4+Foxp3+ Treg in the bone marrow, percentages of BM-residing CD4+Foxp3+ T regulatory cells were markedly higher (p<0.001 and p<0.01) in patients post alloSCT (3.3±0.3%) than in normal BM (1.0±0.3%) or in BM of untreated MM patients (1.8±0.4%). In both groups of patients (p<0.05) as well as in the healthy controls (p<0.001) percentages of Treg were higher in the peripheral blood than in the bone marrow. While there were no differences regarding the percentages of peripheral Treg between the remaining groups, patients post alloSCT had higher percentages of peripheral Treg than newly diagnosed patients (5.6±0.8 vs. 3.2±0.7%, p<0.05). More than 90% of these donor-derived peripheral and BM-residing Treg expressed a memory T cell phenotype, being negative for CD45RA and CCR7. Importantly, peripheral as well as BM-residing Treg of patients post alloSCT were capable of inhibiting the proliferation of autologous non-Treg CD4+ T cells.
CONCLUSION: Our study demonstrates for the first time an increased frequency of immunosuppressive Treg in the bone marrow of MM patients. Remarkably, in our patients these memory-type Treg were all donor-derived and led to an efficient replenishment of Treg in the periphery. These Treg might be necessary for the prevention of graft-versus-host disease in the transplanted MM patients, however, they might also contribute to the failure of an effective graft-versus-myeloma effect in the majority of the patients.
Maternal Haplotype at Time of Banking Is an Effective Strategy to Guarantee the Identification and Traceability of Cord Blood Units.
Authors: Bergamaschi Paola , Perotti Cesare , Viarengo Gianluca , Fante Claudia Del , Parisi Cristina , Marchesi Andrea , Bellotti Laura , Galiazzo Valentina , Pasi Annamaria , Martinetti Miryam , Salvaneschi Laura 
Published: October 2019

Abstract
Stem cell transplantation from unrelated cord blood (CB) donors is nowadays a standard practice for treatment of both malignant and non-malignant haematological disorders. The degree of HLA disparities, the cell dose and the prompt availability of the CB unit strongly influence the donor choice. According to FACT Netcord standards, HLA A, B and DRB1 shall be determined on a pre-cryopreservation sample from each CB. Once a CB unit is identified for potential use, a sample is tested to verify HLA type and confirmation of maternal haplotype is provided. The recipient’s HLA typing and its matching to the donor are confirmed as well prior to CB release for transplant. Quality of the stem cell product represents a basic assumption for the final success of transplant and an essential requirement for CB Banks (CBB). Each CBB shall establish a program of quality assurance that includes identification and traceability. The protection of confidentiality of mother and infant donors and the maintenance of linkage of a particular CB to its mother/neonate pair is of the highest priority. For this purpose each CB is assigned a unique numeric identifier that accompanies the unit in all steps of management and by which it is related to its maternal/infant data and reference samples. At the Pavia CBB, all CB donations are routinely typed for both HLA class I and class II by molecular techniques (low resolution for A and B loci and high resolution for DRB1). The mother haplotype is also assessed by molecular methods at time of banking. Therefore each CB/mother pair is checked before inclusion in the inventory. We retrospectively evaluate the impact of such a strategy on ensuring both identification and linkage of our CB repository. We review the data referring to the CBs stored in our Bank, all typed by LR-DNA for class I loci and by LR/HR-DNA for class II loci. Our inventory consists of 1987 units available for donor selection. In 2 pairs (0.1%) the mother’s typing was found fully disparate as respect to the supposed corresponding CB/neonate. The identification cannot be confirmed leading to discard of the unit. We also review the data pertinent to the 45 units released for transplant by our CBB: in all cases the cord’s confirmatory typing was coherent to the previous typing thus confirming the identity of the product ready for shipment. In our hands, the practice of confirming maternal haplotype at time of banking provides the final evidence of correct assignment and identification before inclusion in the inventory, ensuring the highest quality of the product supplied. This strategy also optimizes the time for the search requests management contributing to prompt availability of the graft.
Large Scale Expansion of Activated T Cells: Defining Procedures and Cryopreservation Methods for Clinical Use as Adoptive Cellular Therapy Post-Transplant.
Authors: Wu Jia-Yan , Webber Susan M. , Hill John M. , Ernstoff Marc , Szczepiorkowski Zbigniew , Meehan Kenneth R. 
Published: October 2019

Abstract
We previously demonstrated a laboratory model of ex vivo expansion of mobilized peripheral blood mononuclear cells (PBMNC) that acquire the CD8+CD56+ NKT cell phenotype and aggressively destroy targets through the NKG2D receptor (Cytotherapy.2006;8:141–8). We developed a large scale culture method for clinical use that allows growth, expansion and cryopreservation of these expanded cells for infusion post-transplant. PBMNC were collected by large volume leukapheresis from healthy donors (n=3). Unselected MNC cells (2.5 × 106/ml) were cultured in Life Cell Bags (Nexell Therapeutics) with serum-free AIM V medium (Invitrogen), IL-2 (Chiron, 1000 IU/ml)and OKT3 (Ortho Biotech, 500 ng/ml) at 37° C and 5% CO2. Six bags were arranged from each collection. Fresh cytokines and medium were added on Days 3 and 5, and the cells were harvested on Day 7. The viability of the ex vivo expanded cell populations was 95% +/− 1.8%. Total MNC cell numbers expanded from 2.5 × 106/ml to 6.4 × 106/ml (2.5 +/− 0.5% fold increase). Phenotypic analyses showed CD3 cells increased from 55% (Day 0) to 94% (Day 7; +/−1.42%); the CD4 cells increased from 30% to 58% (+/− 1.48%); the CD8 cells increased from 11% to 48% (+/−3.98%); the CD56 cells increased from 17.2% to 47.8% (+/− 6.2). Cytotoxicity of Day 7 expanded cells was markedly increased at 74% (+/−7.1%) (Targets: K562 cells; E:T ratio 100:1) compared with 33% (+/− 5.4%) at baseline. We also tested if the ex vivo expanded cells could be cryopreserved for future clinical use. After 7 days in liquid nitrogen, cells were promptly thawed, washed and analyzed. The viability was 91% (+/− 2.1%) and the cytotoxicity against K562 cells was only slightly decreased (65%, +/− 5.4%), when compared to non-cryopreserved expanded cells (p = 0.18). The phenotype was not affected by cryopreservation and thawing. This large scale culture method results in growth and expansion of aggressive effector cells that are cytotoxic against tumor cells. The majority of the expanded cells are CD3+ T cells (equally distributed between CD4+ and CD8+ T cells). NK cells (CD56+) also readily expanded. After cryopreservation of the ex vivo expanded cells and a short-term storage, viability and phenotype of the cells did not change. There is minimal decrease in cytotoxicity that is not statistically significant. This large scale expansion model will be used in clinical trials at our institution to expand cells for adoptive cellular therapy following autologous PBSC transplantation in patients with a hematologic malignancy.
Clinical Efficacy and Feasibility of an Intensive Preparative Regimen with Busulfan, Cyclophosphamide, and Total Body Irradiation Followed by Allogeneic Stem Cell Transplantation for Patients with Myeloid Malignancies.
Authors: Yano Shingo , Asai Osamu , Dobashi Nobuaki , Osawa Hiroshi , Takei Yutaka , Yahagi Yuichi , Takahara Shinobu , Ogasawara Yoji , Otsubo Hiroko , Yamaguchi Yuko , Minami Jiro , Maruyama Dai , Kobayashi Tatsunosuke , Hoshi Yasutaka , Usui Noriko 
Published: October 2019

Abstract
Purpose: High-intensity conditioning regimens before allogeneic stem cell transplantation (all-SCT) could reduce relapse but may increase nonrelapse mortality. We have investigated the efficacy and feasibility of an intensive preparative regimen of busulfan (BU), cyclophosphamide (CY), and total body irradiation (TBI) for patients with myeloid malignancies.
Patients and methods: From November 1990 to February 2005, 80 patients (acute myeloid leukemia (AML) 33, chronic myeloid leukemia (CML) 29, myelodysplastic syndrome (MDS) 18) treated with BU (8 mg/kg), CY (120 mg/kg), and TBI (10 Gy) followed by related (n=46) or unrelated (n=34) allo-SCT. Thirty-eight consecutive patients with AML in 1st remission, CML in chronic phase, and MDS (refractory anemia) defined as a standard risk group, and 42 patients with advanced AML, CML, and MDS (more than 5% marrow blasts) defined as a poor risk group.
RESULTS: With a median follow-up was 6.9 years, the 7-year actuarial overall survival (OS) rate was 58% (standard risk 70% vs poor risk 47%, p=0.0076), and the probability of 7-year relapse rate was 18% (standard risk 17% vs poor risk 18%, p=0.4777). Nonrelapse mortality was 29% (standard risk 26%, poor risk 31%). Multivariate analysis indicated that age older than 40 years (p=0.018) and received allo-SCT before 1998 (p=0.006) had independent predictive value for nonrelapse mortality.
CONCLUSION: The BU- CY- TBI regimen resulted in decreased relapse, especially in poor risk patients. The results of this study suggested that the BU- CY- TBI is an effective and feasible preparative regimen for allo-SCT in patients with myeloid malignancies.
Analysis of Donor Age and Absolute Lymphocyte Count Recovery as Prognostic Indicators of Survival after Allogeneic Blood or Marrow Transplantation.
Authors: Senfield Jeffrey J. , Hahn Theresa , Padmanabhan Swami , McCarthy Philip L. , Battiwalla Minoo 
Published: October 2019

Abstract
Absolute lymphocyte count (ALC) recovery ≥500 cells/mL at day 15 or 30 post-autologous (autoBMT) transplantation has been reported in many studies as an independent prognostic indicator of overall survival (OS) and progression free survival (PFS) in non-Hodgkin lymphoma (NHL), Hodgkin Disease (HD), multiple myeloma (MM) and breast cancer patients. ALC recovery has also been reported in the allogeneic blood and marrow transplant (alloBMT) setting as an important factor in predicting risk of relapse of acute lymphocytic leukemia (ALL) and acute myelogenous leukemia (AML). It has been shown in previous reports that donor age is critically important for OS after alloBMT. In addition, increased donor age correlates with a reduction of common lymphoid progenitor (CLP) differentiation potential which led us to hypothesize that donor age might predict ALC recovery time in related and unrelated alloBMT recipients. We reviewed 76 related and 36 unrelated HLA-matched consecutive patients with hematological malignancies who underwent first alloBMT at Roswell Park Cancer Institute between 4/20/2000 and 3/17/2006. The patient population included 53 males and 59 females with a median (range) age of 44 (4–68) years. Conditioning regimens were FluMel (n=33), CyTBI (n=32), BuCy (n=19), FluCy (n=13), VCT (n=11), and other (n=4). The stem cell source was bone marrow (n=34), peripheral blood (n=77) or both (n=1). Hematologic engraftment was defined as absolute neutrophil count (ANC) reaching 500 cells/ml or more for 3 consecutive days. ALC at day 30 and time to ALC ≥ 500 cells/mL were analyzed. Our results show that peripheral blood (p=0.04) and a higher CD34+ dose (p=0.05) were significantly associated with day 30 ALC ≥ 500. Time to ALC recovery ≥500 cells/mL was not significantly related to donor age for related alloBMT (p=0.340). However, in unrelated alloBMT patients, there was a trend toward increasing donor age associated with a longer time to ALC recovery ≥ 500 cells/mL (p=0.087). ANC recovery correlates with ALC recovery in related (R=0.237, p=0.02) and unrelated (R=0.514, p<0.001) alloBMT recipients. Recipient and donor age were strongly correlated in related alloBMT patients (R=0.89, p<0.001) and weakly correlated in unrelated alloBMT patients (R=0.236, p=0.09). ALC recovery ≥ 500 at day 30 post alloBMT was not a significant predictor of OS in either the related or unrelated subgroups. We will further examine donor-recipient age disparities in unrelated alloBMT patients and present multivariate analyses of predictors for time to ALC recovery.
Reduced Intensity Conditioning Hematopoietic Stem Cell Transplantation (HSCT) with Mesenchymal Stem Cells (MSC) Infusion for Treatment of Metachromatic Leukodystrophy (MLD): A Case Report.
Authors: Meuleman Nathalie , Rocha C. , Bennani Jalil , Tondreau Tatiana , Delforge Alain , Lewalle Philippe , Martiat Philippe , Lagneaux Laurence , Bron Dominique 
Published: October 2019

Abstract
Background: Patients (pts) with MLD have neurological and musculo-skeletal defects with a limited survival. HSCT has been reported as an effective treatment to stabilize or improve defects associated with this disease. Koç and colleagues reported that donor allogeneic MSC infusion is safe and may be associated with reversal of the disease pathophysiology in some tissues (Bone Marrow transplant, 2002). We decide thus to perform a non myeloablative familial HSCT in adult pt with a symptomatic MLD in order to evaluate the safety and the benefit allogeneic HSCT with MSC infusion in a patient with MLD. An informed consent was obtained from the pt.
Patient: a 23 years old women who presented an adult form of MLD for three years was admitted in our department. The most important symptoms associated with the disease were dizziness, proximal weakness of the lower limbs, difficulty to walk, disorder of the memory and urinary incontinence. The reduced intensity conditioning was preferred to decrease the morbidity and mortality of the procedure. MSC were isolated and amplified from the BM-mononuclear cells of the HSC donor. MSC expansion was made in a commercial serum-free medium (UltraCulture, Cambrex, Walkersville, MD) supplemented with a serum substitute (Ultroser, Pall Biosepra, Cergy-Saint-Christophe, France) as previously reported by our group (Eur J Haematol, 2006). Ex vivo expanded allogeneic MSC were intravenously infused at the dose of 1×106 MSC/kg of recipient body weight. The conditioning regimen of the HSCT consisted in Fludarabine and ATG combination and 5× 106 CD34 cells were infused concomitantly with 1.106 MSC cells. We did not observed any immediate or delayed side effects after the MSC infusion. The patient did not presented any complications after the HSCT. At day 8 of the transplantation she had an normal hematological recovery. The platelet nadir was 72.000/mm3 and she did not need any transfusion. With regards of her neurological status, since 22 months the patient had no new deterioration and we observed a stabilisation of the clinical manifestations.
Conclusions: This case report suggests the feasibility and the potential efficacy of reduced intensity conditioning allogeneic HSCT with MSC infusion for patient with MLD. A larger trial is required to confirm this observation.