bluuebunny/crossref_metadata_2025_split huggingface dataset:
A GMCSF & IL15 Fusokine Leads to Paradoxical Immunosuppression In Vivo Via Asymmetrical JAK/STAT Signalling through the IL15 Receptor Complex.
Authors: Rafei Moutih , Wu Jian Hui , Annabi Borhane , Lejeune Laurence , Francois Moira , Galipeau Jacques 
Published: October 2019

Abstract
Immune stimulatory cytokines can be exploited to treat human ailments including cancer. Amongst cytokines identified for such use, granulocyte-macrophage colony stimulating factor (GMCSF) has been under much scrutiny since it acts directly on the adaptive immune system by enhancing antigen presentation as well as costimulation. Furthermore, interleukin (IL)15 possesses overlapping activities with IL2 such as the activation of T-cells and the stimulation of natural killing as well as additive stimulatory effects on the immune system distinct from IL2. These features make IL15 an attractive companion to GMCSF as part of an immunotherapeutic fusokine. Therefore, we hypothesized that a GMCSF and IL15 fusion protein (GIFT15) would possess greater immune stimulatory properties than their combined use. Unexpectedly, tumor cells engineered to secrete GIFT15 protein led to enhanced tumor growth and suppression of natural killer (NK) and NKT-cell recruitment in vivo, suggesting an unheralded immune suppressive effect. We found that GIFT15 has pleiotropic effects on an array of immune competent cells. More specifically, peritoneal macrophages treatment with GIFT15 secrete de novo the tissue inhibitor of metalloproteinase-2 (TIMP-2); activated matrix metalloproteinase-2 (MMP-2); transforming growth factor-β (TGF-β) as well as vascular endothelial growth factor (VEGF). In terms of ligand-receptor interactions, we show by BIAcore analysis that the GIFT15 fusokine has increased affinity for the αchain component of the trimeric IL15R, which contributes to aberrant signalling through the β chain manifested by the hyperphosphorylation of STAT3 both in macrophages and splenocytes. In addition, GIFT15 and IL15R virtual interaction studies suggests that the GMCSF domain component of the GIFT15 fusokine may hinder the interaction of the IL15 domain component with the IL15Rγ chain, and thus leading to a downregulation of the JAK3/STAT5 pathway. We also show that GIFT15 leads to suppression of common γ chain-mediated STAT5 phosphorylation and blockade of the IL15-dependent IFN-γ response in mouse splenocytes We tested the utlity of GIFT15 as an immunosuppressor directly in vivo and demonstrated that it allowed engraftment of allogeneic B16F0 and human xenograft U87MG glioma cells in immunocompetent mice in a CD4-dependent maner. Thus, GIFT15 defines a new class of fusokine which mediates pro-angiogenic and potent immunosuppressive effects via aberrant signalling by the IL15R in lymphomyeloid cells. We propose that GIFT15 may serve as a novel pharmaceutical for tolerance induction of somatic allo or xenografts in mammals without requirement of toxic conditioning regimens.
Generation of Human T/NK Cell Precursors for Adoptive Transfer To Enhance Immune Reconstitution in Allogeneic Hematopoietic Stem Cell Transplantation Recipients.
Authors: Chaudhury Sonali , Zakarzewski Johannes , Shieh Jae-Hung , van der Brink Marcel , Moore Malcolm A.S. 
Published: October 2019

Abstract
Allogeneic hematopoietic stem cell transplantation (HSCT) is associated with significant post-transplant immunoincompetence which affects in particular the T cell lineage and results in an increased susceptibility to infections. Novel strategies to enhance immune recovery after HSCT could prevent malignant relapse and immune deficiency and improve the overall outcome of this therapy. We have established a serum free culture system using murine bone marrow stroma expressing the Notch ligand Delta-like 1 (DL1) to obtain high numbers of human pre-T cells from CD34+ cells. Human cord blood CD34+ cells were plated on OP9 DL1 stroma transduced with adenovirus expressing thrombopoietin (ad-TPO) at an MOI of 30. Media used was QBSF-60 (Serum free media prepared by Quantity Biologicals) supplemented with Flt-3 ligand and IL-7 (10ng/ml). At 4–5 weeks we obtained a 10 5–10 7 fold expansions of cultured cells of which about 70–80% were CD5, CD7 positive pre T cells (Fig 1). We then developed an optimal system to study human lymphohematopoiesis using mouse models (NOD/SCID/IL2rϒnull and NOD/SCIDβ2null) and established an adequate pre T cell number (4 × 10 6) and radiation dose (300 Rads). We injected CD34 and pre-T cells (CD45 +, CD4−, CD5+, CD7+) derived from OP9 DL1 cultures into these mice and achieved ~50%engraftment of NK in the bone marrow and spleen of the mice at 2 weeks following transplant. The thymus from the same mice showed evidence of about 12–15% CD7+ pre T cells. We are currently studying the function of the generated NK and T cells both in vivo and in vitro studies.
Figure Figure
Identification of a Single MiHA Specificity That Induces Resistance to MHC-Matched Allogeneic HCT.
Authors: Shatry Alwi M. , Levy Robert B. 
Published: October 2019

Abstract
T cell populations specific for transplantation antigens have been detected in sensitized individuals following multiple blood transfusions, marrow transplants as well as in multiparous females. Resistance to allogeneic hematopoietic cell transplant (HCT) in such sensitized individuals is consistent with the presence of a host memory T cell (TM) population specific for donor cell antigens. We hypothesized that a single donor minor histocompatibility (MiHA) epitope could elicit antigen-specific CD8 TM capable of resisting MHC-matched allogeneic hematopoietic cell engraftment. To address this question, CD8 TM were generated against a single MiHA epitope to determine if such cells could mediate resistance after ablative TBI conditioning. B6 mice were sensitized 2X to the H60 immunodominant MiHA epitope utilizing bone marrow-derived dendritic cells pulsed with the H60 (LTFNYRNL) peptide. Three weeks following booster sensitization, CD8 T cells were detected by tetramer staining in peripheral blood samples. These T cells exhibited a phenotype characteristic of memory cells (CD44hi, Ly 6C+). B6 (H2b) mice containing CD8+ H60+ T cells were subsequently conditioned with 9.0 Gy TBI and transplanted with 5 × 106 BALB.B (H2b) BM-TCD. One week post-transplant, naive recipients of BALB.B (H60+) or B6-H60 congenic TCD-BM contained >10-fold higher levels of circulating donor cells than the B6 dendritic cell/peptide sensitized recipients. Donor progenitor cells were also found to be significantly reduced in sensitized recipients of allogeneic TCD-BM at this time. Two weeks post-HCT, recipients of syngeneic marrow exhibited >10-fold greater frequency of circulating donor cells compared to recipients of MHC-matched allogeneic marrow (< 5% donor chimerism was detected). These findings demonstrate that host T cells with specificity against a single donor MiHA determinant are sufficient to induce resistance to MHC-matched allogeneic marrow engraftment. Such observations regarding the effector response of HVG contrast those by donor T cell responses post-transplant in which single MiHA differences fail to induce GVHD. Finally, heterologous immunity to virus has been reported to generate allo-reactive TM cells. Since such TM repertoires could include specificity for MiHA immunodominant epitopes, the presence of TM populations that can mediate resistance in “naive” recipients may be more prevalent than hitherto appreciated.
Impaired GvL Potential of Natural Killer Cells Early Reconstituting Following Haploidentical HSCT.
Authors: Vago Luca , Zino Elisabetta , Di Terlizzi Simona , Forno Barbara , Stanghellini Maria T. Lupo , Perna Serena K. , Mazzi Benedetta , Peccatori Jacopo , Rossini Silvano , Roncarolo Maria G. , Bonini Chiara , Ciceri Fabio , Bordignon Claudio , Fleischhauer Katharina 
Published: October 2019

Abstract
Alloreactive NK cells have been suggested to be important functional players in GvL activity after haploidentical HSCT for high risk leukemia. In this study we have characterized NK cells differentiating from purified haploidentical CD34+ cells after transplantation into 16 patients who did (n=8) or did not (n=8) suffer acute leukemia relapse in a long term follow-up (median 208 days). The incidence of relapse in these patients was not correlated with the presence (n=9) or absence (n=7) of predicted donor NK alloreactivity (p=0.94). NK cells in the first month after transplantation were, regardless of the occurence of relapse, NKG2A+ (>95%) and KIR− (13%), thus resembling CD56bright NK cells from healthy donors. However, in contrast to mature CD56bright cells, the patients’ NK cells expressed heterogeneous intensities of CD56, were only partly positive for the lymph node homing markers CD62L and CCR7, and expressed a higher amount of Fcγ receptor III (CD16). Importantly, in contrast to mature CD56bright cells, which constitrutively express the high affinity αβγ IL-2 receptor, thus releasing γ-IFN in response to low dose IL2, the patients’ NK cells lacked IL-R α (CD25) and did not release cytokines in response to low-dose IL2, nor, most importantly, when challenged with leukemic blasts. γ-IFN release induced by leukemic blasts could be restored by inhibition of NKG2A while cytotoxicity, which was consistently lower as compared to that of mature CD56+ cells, could not. Our data suggest that NK cells differentiating in patients from CD34+ progenitors after haploidentical HSCT have important phenotipical and functional differences from both subsets of mature NK cells, accounting for an impaired in vivo GvL potential.
Insertional Activation of MDS1/EVI1, PRDM16 and SETBP1 in a Successful Chronic Granulomatous Disease (CGD) Gene Therapy Trial.
Authors: Schwarzwaelder Kerstin , Schmidt Manfred , Deichmann Annette , Ott Marion G. , Stein Stefan , Glimm Hanno , Siler Ulrich , Hoelzer Dieter , Seger Reinhard , Grez Manuel , von Kalle Christof 
Published: October 2019

Abstract
The potential of gene therapy to correct genetic diseases of the lymphoid compartment has been demonstrated in ADA-SCID and X-linked SCID clinical gene therapy trials. The first successful correction of the myeloid compartment could be achieved in the latest chronic granulomatous disease (CGD) gene therapy trial. CD34+ bone marrow derived cells of 2 patients were transduced using a SFFV based retroviral vector encoding the therapeutic transgene gp91phox. After non-myeloablative conditioning the autologous cells were reinfused. 3 months post therapy the proportion of marked granulocytes was 20% in patient 1 and 10% in patient 2. 5 to 9 months after treatment the proportion of gp91phox expressing granulocytes expanded 4-fold in both patients. Until the latest time points analyzed, (P1: d820; P2: d560) the marking efficiency persisted at that level. In order to define the clonality of the corrected hematopoietic repopulation we accomplished linear amplification mediated PCR (LAM-PCR) on peripheral blood and bone marrow samples as well as sorted lymphoid and myeloid fractions derived from successive time points after therapy. To characterize the retroviral insertion site distribution, we carried out high throughput sequencing and mapping of the vector genome junctions. The hematopoietic repopulation in patient 1 was polyclonal up to day 542 after therapy. Subsequently the number of corrected cell clones and the activity of a predominant clone decreased up to 820 days post transplantation, when the patient succumbed to infectious complications. In this time frame, a different predominant clone appeared. The repopulation in patient 2 has been polyclonal until the latest time point analyzed. Identification of 435 integration sites from patient 1 and 330 insertion sites from patient 2 revealed the gene coding region of the zinc finger transcription factor homologues MDS1/EVI1 and PRDM16 as common integration sites (CIS) in both patients and the SETBP1 locus as a third CIS in patient 1. RT-PCR analysis demonstrated an activating influence of vector LTR on individual CIS genes. Our data show that prospectively studying insertions and stem cell contributions is feasible and that retroviral vector insertion may lead to an upregulation of genes causing an in vivo expansion of the affected cell clones, which can augment gene-corrected hematopoietic repopulation as an unexpected, thus far non-adverse side effect.
Transient Blockade of ICOS Co-Stimulatory Pathways Induces Long-Term Tolerance to Factor VIII Following Nonviral Gene Transfer of Factor VIII Plasmid into Hemophilia A Mice.
Authors: Peng Baowei , Ye Peiqing , Blazer Bruce R. , Ochs Hans D. , Miao Carol H. 
Published: October 2019

Abstract
Formation of inhibitory antibodies is a significant problem encountered in the treatment of hemophilia by replacement therapy. Nonviral gene transfer of a factor VIII plasmid into hemophilia mice induced strong humoral responses through predominantly TH2 signals. The plasmid-treated mice produced persistent, high-level inhibitory antibody specifically against FVIII, representing a unique and useful model for testing various immunomodulation strategies. It was previously demonstrated by our group that transient immunosuppression by CTLA4-Ig and anti-CD40L (MR1) can prevent inhibitory antibody formation following nonviral gene transfer of FVIII plasmid into hemophilia A mice. In this study, we tested if blockade of inducible costimulator (ICOS)-ICOS ligand (ICOSL) pathway in combination with or without agents blocking other co-stimulatory pathways can modulate the immune response following gene therapy treatment. Three groups of mice (n=5/group) were subjected to administration of FVIII plasmid via hydrodynamics-based tail-vein injection, and transient immunosuppressive regimens including anti-ICOS (8 treatment in 2 week period), combination of anti-ICOS (same dose) and CTLA4-Ig (2 treatment at day 0 and 2), and combination of anti-ICOS (same dose) and MR1 (5 treatment in 2 week period). 2 mice from anti-ICOS only group, 3 mice from combination treatment of anti-ICOS and CTLA4-Ig group, and 2 mice from combination treatment of anti-ICOS and MR1 group developed inhibitors at 2 weeks post treatment. The rest of the mice did not develop inhibitors. These results imply that neither synergistic nor additional modulation was achieved by combining CTLA4-Ig or MR1 with anti-ICOS compared to anti-ICOS alone. Subsequently a more frequent and longer anti-ICOS treatment (16 treatment in 4 week period) was administered in two separate groups of FVIII plasmid-treated mice (n=5 and 11 per group, respectively). All the treated mice did not produce inhibitory antibodies against FVIII and produced persistent, high-level (100–300 μg/ml) FVIII gene expression for at least 150 days (experimental period). The CD4+ T cells isolated from the spleen of tolerized mice did not proliferate in response to FVIII stimulation in vitro. Furthermore, higher population of CD4+CD25+ regulatory T cells were detected in peripheral blood in the tolerized mice compared to untreated and plasmid-treated mice. Adoptive transfer of CD4+ T cells isolated from tolerized mice is performed to test if these cells can protect the recipient mice from developing inhibitory antibodies against FVIII. Anti-ICOS treatment has the potential for a new immunomodulatory strategy for preventing the formation of inhibitory antibodies against FVIII following gene therapy.
Using Geographical Information Systems (GIS) To Examine Associations between Characteristics of Males with Hemophilia and Geographic Distance to Hemophilia Treatment Centers (HTCs).
Authors: McClellan Ann C. , Soucie J. Michael , Kulkarni Roshni 
Published: October 2019

Abstract
Hemophilia, an inherited bleeding disorder, is best treated at specialized healthcare facilities offering comprehensive healthcare services. For healthcare providers at the over 130 federally-funded HTCs across the U.S., information about where individuals with hemophilia live and distances to the HTC can be useful in studying barriers to care and health outcomes. For the analysis, we used data collected on males with hemophilia in the CDC-sponsored Universal Data Collection (UDC) surveillance program that includes demographic and clinical data on about 90% of patients using these HTCs for their care. Patient characteristics, including age, race/ethnicity, insurance status, hemophilia severity, treatment (prophylaxis vs. on-demand) and residential 3-digit zip code were obtained from the UDC database. We calculated distances between the HTC and the centroid (geographic center) of the patients’ zip codes using GIS software (sample map shown in figure 1). The independent associations between patient characteristics and distance to the HTC adjusting for regional variations were examined using a multivariate generalized linear model. The mean age of the patients was 21 years (median 16 years); 70% were white; 42% used a health maintenance organization (HMO), 27% had Medicare or Medicaid coverage, and 16% had commercial health insurance; 50% of patients had severe hemophilia; and 26% were on prophylaxis. The mean distance from patient residence to HTC was 68 miles (median 35 miles). Of the 12,397 patients living in the contiguous U.S., 10,164 (82%) lived within 90 miles of the HTC (53% within 30 miles, 31% from 31–60 miles, and 16% from 61–90 miles). Among patients who lived within 90 miles of the HTC, we found that patients >25 years old and those on prophylaxis lived closer to HTCs than their counterparts (p=0.04 for each). On the other hand, white patients and those using an HMO lived farther from HTCs than those of other races and those with other types of health insurance, respectively (p<0.001 for each). Information from this geographic analysis may be used to guide resource allocation and programmatic activities and may help during emergencies such as natural disasters and treatment product shortages.
North Carolina Hemophilia Treatment Centers Male UDC* Participants with Hemophilia North Carolina Hemophilia Treatment Centers Male UDC* Participants with Hemophilia
RADAR Update on Thalidomide (Thal)- and Lenalidomide (Len)-Associated Venous Thromboembolism (VTE): Safety Concerns Persist for Multiple Myeloma (MM) Despite FDA Approvals in This Setting.
Authors: Bennett Charles L. , Hussain Zehra , Courtney Mark , Yarnold Paul , Raisch Dennis , McKoy June M. 
Published: October 2019

Abstract
BACKGROUND: At ASH 2004, we reported > 20% VTE rates with thal/dexamethasone (thal/dex) use as an off-label treatment for MM. The abstract prompted Richard Blumenthal, the Connecticut Attorney General, to file a Citizen’s Petition with the FDA in 2005 outlining six recommendations to address this safety concern. In 2006, the FDA indicated that four recommendations were approved. The AG’s request for a randomized clinical trial of aspirin, low molecular weight heparin (LMWH), and warfarin as VTE prophylaxis was denied- with the FDA indicating that preliminary data suggested that these agents are effective for prophylaxis. At ASH 2005, we reported > 20% VTE rates with len/dex treatment for MM. In response, the manufacturer pre-emptively included a “Black Box” warning describing high VTE rates when len was used off-label for MM. In 2006, len and thal received FDA approval for MM. The Research on Adverse Drug Events and Reports (RADAR) project conducted a systematic review of VTE rates when len or thal is prescribed for MM.
METHODS: Reports of VTE with thal or len treatment of MM were reviewed. Data sources included: the FDA’s MedWatch program and phase II/III clinical trial reports from 1998 to 7/2006.
RESULTS: FDA’s MedWatch program included reports of VTE occurring among 8 len- (3 on aspirin, 2 on warfarin, and 1 on MLWH) and 1076 thal-treated cancer patients. Clinical trials identified VTE rates of 15% among thal/dex treated patients and 14% among len/dex- treated MM patients (Table). Uncontrolled trials found that thal/dex-associated VTE rates were sometimes, but not always, lower with aspirin, LMWH, or warfarin (INR of 2 to 3), while little information on VTE prophylaxis is available for len-dex treated MM patients.
CONCLUSIONS: The extant data continue to support our assertion that randomized controlled trials are needed to identify appropriate VTE prophylaxis agents when MM patients receive len or thal with dex. Data on aspirin prophylaxis remain premature.
Clinical Trial VTE Results VTE rate among Thal treated MM patients (n = 2690) VTE rates among Len treated MM patients (n = 702) MM-extent of disease Untreated Relapsed/Refractory Untreated Relapsed/Refractory n/a = data not available     Thal/Dex None 96/667 (14.4%) 11/256 (4.3%) 38/278 (13.7%) 48/346 (13.9%) Warfarin (1–1.25 mg/d) 16/105 (15.2%) 8/85 (9.4%) n/a n/a Aspirin (81 mg/d) n/a n/a 9/54 (16.7%) n/a LMWH/Warfarin (INR 2–3) 0/63 (0.0%) 2/26 (7.7%) n/a n/a     Thal/Dex/Doxorubicin None 148/610 (24.3%) 47/271 (17.3%) n/a n/a Aspirin (81 mg/d) 3/45 (6.7%) n/a n/a n/a LMWH 19/211 (9.0%) n/a n/a n/a     Thal/Melphalan/Prednisone None 43/287 (15.0%) n/a n/a n/a Aspirin (81 mg/d) n/a n/a 1/24 (4.2%) n/a LMWH 2/64 (3.1%) n/a n/a n/a
Darbepoetin Alfa (DA) - Pattern of Use and Outcomes in Adult Patients with Non-Myeloid Malignancies and Chemotherapy-Induced Anemia: A French Observational Study.
Authors: Pigneux Arnaud P.M. , Ray-Coquard Isabelle L. , Dalivoust Philippe J.M. , Biostat Gaelle Chenuc , Bastie Anne M. 
Published: October 2019

Abstract
INTRODUCTION: DA is widely used to treat anemia in patients undergoing chemotherapy (CT) for non-myeloid malignancies. Three years after the commercialization of DA in France, a study of the pattern of DA use in daily practice was conducted in this population.
METHODS: A nationwide, retrospective observational study of DA use and patient outcomes was conducted in 239 centers between September 2004 and May 2005. Adult patients with non-myeloid malignancies, who had received DA for CT-induced anemia for at least three 3- or 4-weekly or at least five 2-weekly CT cycles (min. 9–12 weeks) were studied. Response was defined as hemoglobin (Hb) ≥ 12g/dL or a Hb increase of at least 2 g/dL from baseline to week 6/8. In addition, the proportion of patients achieving a Hb level of ≥ 11 g/dL as suggested by NCCN guidelines was assessed. Hb levels were evaluated at baseline, week 3 and week 6 (CT cycles of 21 days) or week 4 and week 8 (CT cycles of 15 or 28 days).
RESULTS: Of 2017 patients recruited, 1865 (92.5%) were evaluable at week 6 or 8. Analyses were conducted on the evaluable patient population. A total of 51% were female and the mean age was 62.5 years [range: 18–96 years]. The most common tumor types were non-myeloid hematological malignancies (25%), lung cancer (21%), breast cancer (16%) and gastrointestinal tumors (14%). A total of 37% of centers were University hospitals, 30% public hospitals, 21% private hospitals and 11% specialized cancer centers. Baseline mean Hb level was 10g/dL [range 5–16.6 g/dL]. A total of 843 patients at week 0 (45.4%) had a Hb <10g/dL. The first DA dose was administered to 40% of patients at the first CT cycle and to 60% of patients at subsequent cycles. At week 6/8, 707 patients (38%) had a Hb ≥ 12 g/dL and 1096 patients (58.8%) had a Hb ≥ 11 g/dL. During the study period, between week 0 and week 6/8, 272 pts (15.9%) received RBC transfusions.
CONCLUSIONS: Despite the very short follow-up of 6/8 weeks, almost 40% of the patients achieved a Hb ≥ 12 g/dL. More importantly, the proportion of patients reaching a Hb level within the range recommended by NCCN guidelines (≥ 11 g/dL) at this short follow-up was around 60%, which is comparable to results published in other trials of DA.