bluuebunny/crossref_metadata_2025_split huggingface dataset:
Methylprednisolone Versus Intravenous Immunoglobulin As Initial Therapy In Adult Immune Thrombocytopenia
Authors: Song Ik-Chan , Jo Deog-Yeon , Choi Yoon Seok , Lee Hyo-Jin , Yun Hwan-Jung , Kim Samyong 
Published: October 2019

Abstract

Background
Both intravenous methylprednisolone (methyl-Pd) and intravenous immunoglobulin (IVIg) have been used as standard initial therapies in adult patients with immune thrombocytopenia (ITP). Nonetheless, few studies have addressed whether there are differences in clinical efficacy between these two treatments. In this study, we compared platelet responses, toxicities, and bleeding-related mortality associated with these two treatments in adult ITP.

Patients and Methods
Adult patients newly diagnosed with ITP, with platelet counts of less than 20,000/mL, were prospectively enrolled. Patients with comorbidities such as diabetes and uncontrolled cardiovascular disorders and who were pregnant were excluded. Patients who entered the study between 1993 and 2002 received intravenous methyl-Pd therapy (10 mg/kg/day for 3 days) followed by oral Pd (1 mg/kg/day). Patients who entered the study between 2003 and 2008 received IVIg (400 mg/kg/day for 5 days) together with oral Pd (1 mg/kg/day). After 6 weeks, the prednisolone began to be tapered in both arms. Early response and maintenance of response were assessed at 7 days and 6 months after treatment, respectively, based on the outcome criteria proposed by an international working group (Blood 2009).

Results
Of 87 patients enrolled, 77 (88.5%) were eligible for analysis. Initial platelet counts did not differ between the two groups. Early responses occurred in 30 of 39 patients (76.9%) receiving methyl-Pd versus 33 of 38 patients (86.6%) receiving IVIg (p = 0.187). Maintenance of response was observed in 28 patients (71.8%) in the methyl-Pd arm and 23 patients (60.5%) in the IVIg arm (p = 0.187). The time to a complete response in the IVIg arm (6 days; range, 1–35 days) was shorter than that in the methyl-Pd arm (13.5 days; range, 2–29 days) (p = 0.002). Side effects were mild and tolerable in both arms. Five years after treatment initiation, 5 patients (12.8%) and 7 patients (18.4%) were still under salvage treatment in the methyl-Pd and IVIg arms, respectively. During a median follow-up of 98 months (range, 33-228 months) in the methyl-Pd arm and 71 months (range, 52-180 months) in the IVIg arm, 19 patients (48.7%) and 18 patients (47.4%) patients required one or more salvage treatments, respectively (p = 0.906). No bleeding-related death was observed in either group during the follow-up period.

Conclusion
These results indicate that neither the early response rate nor the long-term outcome differs between methyl-Pd and IVIg treatments. IVIg, however, more rapidly induces a complete response than methyl-Pd.

Disclosures:
No relevant conflicts of interest to declare.
A Carboxy-Terminally Truncated G-CSF Receptor Abolishes Apoptosis Induction By Neutrophil Elastase G185R Mutant In Myeloid Cells,
Authors: Dong Fan , Qiu Yaling , Hu Nan , Friedman Alan D. 
Published: October 2019

Mutations in ELANE encoding neutrophil elastase (NE) have been identified in the majority of patients with severe congenital neutropenia (SCN). SCN patients are predisposed to acute myeloid leukemia (AML), and progression from SCN to AML is accompanied by acquisition of mutations in CSF3R encoding the G-CSFR receptor (G-CSFR) in approximately 80% of the patients. The mutations, which cause receptor’s carboxy-terminal truncation, are highly specific for SCN/AML, but the reason for the prevalence of the CSF3R mutations in patients with SCN/AML is unknown. We show here that NE expression was induced by G-CSF in myeloid 32D cells expressing the wild type (WT) G-CSFR (32D/GR), but not in cells expressing an SCN-derived truncated G-CSFR, d715 (32D/d715). Unlike the WT G-CSFR, the d715 receptor was unable to activate a 1.8-kb promoter fragment of mouse Elane. The NE mutants associated with SCN have been shown to cause premature apoptosis of differentiating granulocytic precursors. To address whether the d715 receptor might abolish apoptosis induction by the NE mutants in part by suppressing their expression, we placed the cDNA encoding the SCN-derived NE G185R mutant under the control of the Elane promoter and the resultant expression construct was transfected into 32D/GR and 32D/d715 cells. G-CSF treatment resulted in increased expression of G185R mutant and premature apoptosis in 32D/GR cells, but had no effect on G185R expression and cell survival in 32D/d715 cells. We further addressed whether the d715 receptor was capable of directly suppressing apoptosis induced by the G185R mutant. As expected, forced expression of the G185R mutant in 32D/GR cells led to accelerated apoptosis in response to G-CSF, but did not negatively influence the viability of 32D/d715 cells cultured in G-CSF. To evaluate whether the d715 receptor inhibited apoptosis induction by the G185R mutant in other hematopoietic cells, we expressed the G185R mutant in murine multipotent FDCP-mix A4 cells transfected with WT G-CSFR (FDCP/GR) or d715 receptor (FDCP/d715). Although FDCP/GR cells expressing G185R mutant showed no significant increase in apoptotic cell death upon treatment with G-CSF, they exhibited increased sensitivity to tunicamycin-induced endoplasmic reticulum (ER) stress. In contrast, the G185R mutant failed to sensitize FDCP/d715 cells to apoptosis induction by tunicamycin. Thus, it appears that the d715 receptor may abolish G185R mutant-induced apoptosis by two distinct mechanisms, i.e., suppression of its expression and direct inhibition of its pro-apoptotic action. Our data may have implications for understanding the prevalence of CSF3R mutations in patients with SCN/AML. We propose that the CSF3R mutations might represent a mechanism by which myeloid cells carrying the ELANE mutations evade the pro-apoptotic effect of the NE mutants in SCN patients.

Disclosures:
No relevant conflicts of interest to declare.
Caveolin-1 Is Required For T Cell-Mediated Acute GvHD
Authors: Wohlfeil Sebastian , Anne-Kathrin Hechinger , Leonhardt Franziska , Prinz Gabriele , Schmitt-Gräff Annette , Minguet Susana , Zeiser Robert 
Published: October 2019

Caveolin-1 (Cav-1) is a key organizer of membrane specializations that coordinate membrane and protein traffic in different cells including T cells. Cav-1 promotes the formation and stability of plasma membrane raft microdomains that serve as platforms for signal transduction and binds a wide array of signal transducers through interactions with its phosphorylated tyrosine 14 or the so-called scaffolding domain. Several of the proteins identified as Cav-1 binding partners have been suggested to play a role in TCR-regulated membrane dynamics and intracellular signaling. Therefore, we aimed to determine the requirement of Cav-1 for T cells during acute graft-versus-host disease (GvHD).
We observed upregulation of Cav-1 in the T cells of mice developing acute GvHD which was dependent on both tissue damage and homeostatic signals as investigated in conditioned WT recipients or RAG-/-C gamma chain-/- non-conditioned mice. By using gene targeted mice we found that Cav-1 deficiency of the donor led to improved survival (p=0.02), lower GvHD histopathology scores (p=0.004) and reduced serum levels of IL-6 (p=0.02) while Treg numbers increased when Cav-1 deficient donors were used. Comparative microarray analysis of WT compared to Cav-1 deficient T cells indicated a reduced IL-17A production in Cav-1 T cells exposed to allogeneic dendritic cells. These observations were confirmed on the protein level.
In summary we show a role for Cav-1 in T cells during GvHD as an alloantigen driven T cell response. Regarding the mechanism we describe the relevance of Cav-1 for Th-17 lineage commitment of alloreactive T cells during GvHD.

Disclosures:
No relevant conflicts of interest to declare.
TLR7 Ligand Potentiates GM-CSF-Initiated Antitumor Immunity Through Activation Of Plasmacytoid Dendritic Cells
Authors: Narusawa Megumi , Inoue Hiroyuki , Sakamoto Chika , Matsumura Yumiko , Takahashi Atsushi , Watanabe Ayumi , Inoue Tomoko , Miyamoto Shohei , Takayama Koichi , Inoue Makoto , Hasegawa Mamoru , Nakanishi Yoichi , Tani Kenzaburo 
Published: October 2019

Vaccination with irradiated granulocyte macrophage-colony stimulating factor (GM-CSF)-transduced autologous tumor cells (GVAX) has been shown to induce therapeutic antitumor immunity presumably through the activated maturation of myeloid dendritic cells (DCs). However, its effectiveness is limited, and little is known about the biological properties related to GM-CSF-sensitized DCs (GM-DCs) in tumor-draining lymph nodes (TDLNs). We therefore attempted to enhance the antitumor effect of GVAX therapy by identifying the key pathways in GM-DCs in TDLNs. We initially confirmed that syngeneic mice subcutaneously (s.c.) injected with poorly immunogenic Lewis lung carcinoma (LLC) cells transduced with Sendai virus encoding GM-CSF (LLC/SeV/GM) significantly rejected the tumor growth. Using cDNA microarrays, we obtained a large number of gene expression data from CD86+ GM-DCs and control DCs in TDLNs, and found that the expressions of type I interferons (IFNs)-related genes, including IRF7 and TLR7, known to be abundantly expressed in plasmacytoid DCs (pDCs), were upregulated in GM-DCs. Indeed, to further activate pDCs, mice s.c. challenged with LLC/SeV/GM cells in combination with a TLR7 ligand, imiquimod, but not a TLR4 ligand, LPS, significantly suppressed the tumor growth compared with mice treated with LLC/SeV/GM cells alone. In contrast, pDCs-depleted mice challenged with LLC/SeV/GM cells facilitated the tumor growth, strongly suggesting that pDCs are essential immune subpopulation in exerting GM-CSF-initiated antitumor effects.
Furthermore, the additional use of imiquimod overcame the refractoriness of therapeutic vaccines with irradiated LLC/SeV/GM cells in mice with pre-established LLC tumors. Moreover, similar improvement of GVAX therapy was also observed in a mouse model of CT26 colorectal cancer. Mechanistically, mice treated with the combined vaccination displayed increased cell ratio of PDCA-1+ pDCs and expression levels of CD86, CD9, which correlate with an antitumor phonetype, and Siglec-H, which promote CD8+ T cell proliferation, in TDLNs. Indeed, allogeneic MLR test showed that bone marrow-derived pDCs matured by in vitro culture with GM-CSF plus imiquimod elicited a superior capacity of CD8+ T cell proliferation compared with those with GM-CSF only. On the other hand, the ratio of CD4+CD25+FoxP3+ regulatory T cells (Tregs) was decreased in TDLNs mice treated with the combined vaccination.
These findings collectively elucidated that pDCs play positive roles in GM-CSF-initiated antitumor immunity and that further activation of pDC by imiquimod targeting TLR7-IRF7 dependent type I IFNs pathway enhance the antitumor effects of GM-CSF-based tumor vaccination.

Disclosures:
No relevant conflicts of interest to declare.
Alternative Donor Stem Cell Transplantation In Aplastic Anemias and Refractory Cytopenias Is Save and Feasible With T- and B-Cell Depleted Haploidentical Grafts
Authors: Feuchtinger Tobias , Teltschik Heiko-Manuel , Pfeiffer Matthias , Mueller Ingo , Schrauder Andre , Handgretinger Rupert , Lang Peter 
Published: October 2019

Allogeneic stem cell transplantation is still the curative treatment option for the majority of patients with severe aplastic anemia and refractory cytopenias. However, a HLA-matched donor is not available for all patients. Alternative donor transplantation has been an experimental treatment option, limited by high rejection rates and transplant related mortality. We performed a prospective clinical trial to evaluate the safety and feasibility of haploidentical stem cell transplantation, since haploidentical family members are always available donors. We investigated a cohort of 10 pediatric patients with severe aplastic anemia or myelodysplatic syndrome transplanted (refractory cytopenia) with T-cell depleted grafts between 2004 and 2011. 7 patients had myelodyslastic syndrome with refractory cytopenia (MDS-RC), 3 had severe aplastic anemias (SAA) refractory to immunosuppresive treatment. 3 patients received a 2nd SCT after rejecting the graft from matched donors. Median age was 11.4 years. Standard conditioning regimen consisted of Fludarabin 3-4x40mg/m2, Thiotepa 1-3x5mg/kg, Melphalan 2x70mg/m2 (n=8) and serotherapy using OKT3 (n=5) and ATG (n=5). 8 patients received additional total lymphoid irradiation (TLI 7 Gy) to prevent graft rejection. In vitro graft manipulation was carried out by direct depletion using antiCD3/19 magnetic microbeads. A median number of 10.1x106 CD34+ progenitor cells and 27x103 T-cells/kg body weight (BW) were transfused. Pharmacological GvHD prophylaxis (graft vs. host disease) was carried out with Mycofenolate until day 60, if residual T-cells in the graft exceeded 25000/kg BW. Primary engraftment occurred in all patients Median time to reach 500/µl neutrophiles was 9 days (9-11). Independence from platelet substitution was reached after 13 days (8-16). Three patients rejected the graft later on. 6/10 patients had no signs of acute GvHD or GvHD grade I, 2 patients had GvHD grade II. TRM at day +100 and after 1 year was 0% and 20%, respectively. Event free survival (EFS) at 3 years was 80%.

Conclusions
Haploidentical SCT with T-cell depleted grafts is a therapeutic option for refractory cytopenias and severe aplastic anemia after nonresponse to immunosuppressive treatment if no HLA-matched donor is available. Recovery of neutrophiles and platelets were fast and TRM was low, even if retransplantation was necessary. Since spontaneous outcome of these conditions are poor, alternative donor SCT is a realistic option for these patients.

Disclosures:
No relevant conflicts of interest to declare.
Differential Gene Expression Analysis Predicts For Response To Bortezomib In The Treatment Of Steroid Refractory Chronic Graft-Versus-Host Disease (cGVHD)
Authors: Miller Alan M , Schneider Reva , Vance Estil A. , Agura Edward , Powell Sheila , Pascual Virginia , Munagala Indira , Qin Huanying 
Published: October 2019

We conducted a trial of bortezomib in patients with steroid-refractory cGVHD. Patients received 1.6 mg/m2 q wk x 4 followed by one week of rest, for up to six cycles, with some going on to a maintenance phase of treatment every other week for an additional 6 months. Twenty-one patients were registered on trial two patients withdrew before beginning treatment. Nineteen patients received between two and twelve months of treatment. The treatment was well tolerated with no grade 3 or higher adverse events (AE). Six patients had either CR or near CR. Average Rodnan scores on the 8 patients with sclerosis completing 2 or more cycles of therapy decreased from 22.6 ± 12.8 to 5.9 ± 6.2, p<0.005. One patient with non-healing suppurating lesions of his lower extremities had marked healing, another had significant decrease in chronic diarrhea. Weekly bortezomib for cGVHD was well tolerated and resulted in early improvement in a subset of patients with long standing refractory disease.
Differential gene expression analysis was performed on the blood of patients prior to starting therapy, after two cycles and after their final cycle of therapy. All samples were run simultaneously at the end of the trial. Patients were classified as High (CR or near CR), intermediate (PR or stable disease), or poor responders (progression or not evaluable) based on clinical response. At baseline cGVHD patients express an overall decrease in T cell markers such as lineage (CD4), chemokine receptors (CCR6, CXCR3 and CCR7), and co-stimulatory molecules (CD40L, ICOS, CD28, and CD27), as well as decreases in IL23 and Granzyme K message compared to healthy controls. Increases were seen in myeloid/macrophage markers (CD163 and OSCAR). Baseline profiles of good responders were closer to those of healthy controls as compared to poor responders (figure below). Inflammation, cell death and apoptosis modules are up regulated and immune response modules are down-regulated in low and medium responders
HC – Healthy Control; DC – Disease Control (allogeneic BMT recipients without cGVHD)
Weekly bortezomib for cGVHD appears to be well tolerated and results in early improvement in long standing refractory disease in a subset of patients. Gene expression profiles at the initiation of treatment correlated with response and may be useful in predicting which patients will respond to bortezomib.

Disclosures:
Miller: Millennium Pharmaceuticals: Research Funding. Off Label Use: Bortezomib for chronic graft vs. host disease.
Palifermin For Prevention Of Oral Mucositis Has No Negative Effect On Long-Term Outcome In Patients With Hematological Malignancies Undergoing HSCT - Long-Term Follow-Up To 15 Years
Authors: Stiff Patrick J , Spielberger Ricardo 
Published: October 2019

Background
Palifermin has been shown to reduce the incidence and duration of severe oral mucositis in hematological stem cell transplantation (HSCT) recipients. A follow-up study was performed to rule out a potential long-term safety risk of palifermin use.

Objectives
to compare long-term disease outcome (overall survival, disease progression and incidence of secondary malignancies) between palifermin and placebo.

Methods
This is a long-term follow-up study of patients with hematological malignancies undergoing HSCT and treated with palifermin or placebo to prevent oral mucositis. The aim of the study was to detail any potential late complications due to palifermin exposure. Patients were enrolled between 1997-2003 into four randomized, placebo-controlled phase II/III studies conducted at 31 sites in Australia, Europe and the US. The survival outcomes were compared using hazard ratios (HRs) estimated with Cox model including the treatment group, baseline age, disease type (Hodgkin lymphoma, non-Hodgkin lymphoma, multiple myeloma, leukemia), ECOG performance status, country and presence of prior radiotherapy as covariates. The incidence of secondary malignancies was compared with chi-squared test.

Results
A total of 672 patients were randomized to the parent studies (429 palifermin, 243 placebo) and 543 (345, 198) were enrolled to the long-term follow-up. The median follow-up time for subjects alive was 7.9 years (range 0.1-14.9) for palifermin and 8.8 (0.1-14.8) for placebo. No significant differences were seen for either overall survival (HR = 1.01; 95% confidence interval (CI) 0.78-1.31; p=0.921) or progression-free survival (HR=1.04; 95% CI 0.83-1.31; p=0.733). Secondary malignancies were reported by 13% (palifermin) vs. 11% (placebo) of the patients (p=0.477). The most common secondary malignancies were acute myeloid leukemia/myelodysplastic syndrome (5% vs. 5%) and skin cancers (2% vs. 2%).

Conclusion
The overall survival, progression-free survival, and the incidence of secondary malignancies were comparable between palifermin and placebo in patients undergoing HSCT. After a follow-up of up to 15 years, no negative effect of palifermin on long-term outcomes was observed.

Disclosures:
No relevant conflicts of interest to declare.
Estimation Of Serum Erythropoietin Levels in Anaemic HIV Infected Patients in Lagos, Nigeria
Authors: Olu-Taiwo Adebola , Akanmu Suleimon , Amusu Olatilewa , Ogunro Samuel 
Published: October 2019

Background
Erythropoietin(EPO) response to anaemia has been reported to be suboptimal in HIV infected anaemic patients.

Objectives
To assess the relationship between EPO levels and degree of anaemia, CD4+ cell count and viral load of HIV infected treatment naive subjects and if there is any association with inflammatory cytokines.

Subjects and Methods
A total of 120 subjects aged 18-66 years, comprises (1) HIV infected treatment naive subjects who had anaemia with Hb level <10g/dL. Control subjects were: (2) HIV infected non-anaemic, (3) non-HIV infected anaemic and (4) non-HIV infected non-anaemic individuals. Full blood count using automated counter; CD4+ cell count by semi-automated flow cytometer; and viral load were estimated. Serum erythropoietin levels, serum ferritin levels, CRP levels, TNF-α levels and IL-6 levels were estimated. Appropriateness of EPO response was determined by calculating EPO response ratio.

Results
Majority (72%) of the HIV infected anaemic patients had a normocytic hypochromic type of anaemia. A weak positive correlation between EPO levels and CD4+ cell count (r=0.075, p=0.572) was obtained. A significant negative correlation between EPO levels and haemoglobin levels (r=-0.268, p=0.003). Twenty-seven (71.05%) of the 38 HIV infected anaemic subjects had EPO response ratio less than 0.8. Serum ferritin level was significantly higher in HIV infected anaemic subjects compared to the controls (p=0.000). Serum interleukin-6 level was significantly higher in HIV infected anaemic subjects compared to HIV infected non-anaemic subjects (p=0.003). Serum C-reactive protein level was significantly higher in HIV infected anaemic subjects compared to non-HIV infected non-anaemic (p=0.004) and non-HIV infected anaemic subjects and(0.001). Serum tumour necrosis factor-α level was significantly higher in HIV infected anaemic subjects compared to other arms of the study population (p=0.000).

Conclusion
The present study revealed that HIV infected anaemic subjects had reduced serum EPO levels. However, serum inflammatory cytokines levels were raised in the same subjects.

Disclosures:
No relevant conflicts of interest to declare.
Pulmonary Hypertension in Sickle Cell Disease: Correlation With Echocardiographic Assessment and Serum Brain Natriuretic Peptide Concentration
Authors: Alkindi Salam , Jabeen Zeba , Pathare Anil , Al-Huneini Mohammed , Al Khabori Murtadha , Al-Farsi Khalil , Al Riyami Adil , Al-Raesi Majdah , Dennison David 
Published: October 2019

Background & Purpose
Pulmonary hypertension (PH) in sickle cell disease (SCD) is an important risk factor for complications including sudden death. In this study, we aimed to determine the prevalence of PH and correlate the echocardiographic parameters and general laboratory data with markers of hemolysis and serum NT pro-brain natriuretic peptide [BNP] concentrations in Omani SCD patients.

Methods
A cohort of 163 SCD patients [Mean age 25.4±8.4 years], in steady state was prospectively screened for PH with Doppler echocardiography (defined as a tricuspid regurgitation jet flow velocity of ≥ 2.5 m/sec and/or mean pulmonary artery pressure [mPAP] ≥25 mmHg). After a written informed consent, all patients were investigated with a complete blood count, renal chemistry, hemolytic parameters including LDH, haptoglobin, liver function tests, coagulation studies, HPLC studies including HbS and HbF level estimations, X ray Chest, ECG, ABG, Pulmonary function tests, Pulse oximetry, and Serum NT pro-BNP levels.

Results
In the evaluable patient cohort of 116 subjects [63 females, 53-males], the prevalence of PH was 5.2%. No statistically significant differences were detected in Hb levels, ECG, chest radiography, pulmonary function tests between patients with and without PH. However, plasma NT pro-BNP levels were significantly correlated with PAH [r=0.934, p<0.000], TRV jet [r=.671, p<0.000], Abnormal ECHO [r=0.672, p<0.000], and direct Bilirubin levels [r=0.278,p<0.009] in SCD patients. Furthermore, in SCD patients with PH, there was a statistically significant increase in plasma NT pro-BNP levels [ p<0.001], ALT [p<0.02], S. Creatinine [p=0.045] and Total bilirubin [p<0.0001] and direct Bilirubin levels [p<0.0001][Table].

Conclusions
Serum NT pro-BNP is a strong indicator of PH in SCD patients. Doppler ultrasound echocardiography is a useful initial screen for PH in SCD patients. The correlation between PH and hemolytic markers suggests an implication in the pathogenesis of PH.

Disclosures:
No relevant conflicts of interest to declare.