bluuebunny/crossref_metadata_2025_split huggingface dataset:
Reconstitution of STAT4 Restores Defective Interferon-gamma Production and Allows Normal IFN-gamma-Dependent Responses after Autologous Stem Cell Transplantation.
Authors: Robertson Michael J. , Chang Hua-Chen , Pelloso David , Kaplan Mark H. 
Published: October 2019

Abstract
Production of IFN-gamma is critical for optimal antitumor immune responses in several preclinical animal models. Patients with lymphoma who have undergone autologous hematopoietic stem cell transplantation exhibit profoundly diminished IFN-gamma production in vivo during IL-12 therapy (Clin Cancer Res.2002; 8: 3383). Furthermore, post-transplant patient peripheral blood mononuclear cells (PBMCs) stimulated directly in vitro with IL-12 secrete little if any IFN-gamma. To determine if restoration of IFN-gamma production would be sufficient to reconstitute IFN-gamma-dependent responses after transplantation, the integrity of IFN-gamma signaling was assessed in post-transplant patient PBMCs. Compared to control subject PBMCs, post-transplant patient PBMCs expressed equivalent levels of CD119 (IFN-gamma receptor alpha) and STAT1. Moreover, tyrosine phosphorylation of STAT1 in response to IFN-gamma did not differ in post-transplant patient as compared to control PBMCs. Thus, IFN-gamma signaling is intact after transplantation. Our prior studies have shown that STAT4 protein levels are decreased by ~97% in post-transplant patient PBMCs, whereas levels of Jak2, Tyk2, and STAT3 are similar to control PBMCs (Blood2005; 106: 963). We wished to demonstrate directly that STAT4 deficiency is the mechanism of defective IFN-gamma production after autologous transplantation. Enriched CD4+ T cells were isolated from post-transplant patient and control PBMCs and cultured under conditions that promote Th1 differentiation. STAT4 deficiency persisted in post-transplant patient CD4+ T cells cultured under Th1 conditions, whereas these cells expressed normal levels of T-bet. Compared to control Th1 cells, post-transplant patient CD4+ T cells cultured under Th1 conditions exhibited markedly reduced IFN-gamma production after restimulation with CD3 mAb. These results are consistent with expected impairment of Th1 differentiation due to profound STAT4 deficiency. STAT4 cDNA was transiently transfected into Th1-cultured CD4+ T cells from post-transplant patients. After stimulation with CD3 mAb plus IL-12, the amounts of IFN-gamma secreted by STAT4-transfected patient cells were equivalent to or higher than those secreted by control subject cells. These data indicate that reconstitution of STAT4 expression is sufficient to restore IFN-gamma production by post-transplant patient PBMCs. As IFN-gamma signaling is normal post-transplant, circumventing STAT4 deficiency should therefore restore IFN-gamma-dependent antitumor immune responses after autologous transplantation. Future studies will determine the molecular mechanisms of STAT4 deficiency after transplantation and develop clinically feasible methods to circumvent this deficiency.
A Novel Mechanism in Generation of Self-Antigen Repertoire - An Unconventional Antigen Translated by a Novel Internal Ribosome Entry Site Elicits Anti-Tumor Humoral Immune Responses.
Authors: Yang Xiao-Feng , Xiong Zeyu , Liu Enli , Yan Yan , Silver Richard T. , Verstovsek Srdan , Prchal Josef , Wang Hong 
Published: October 2019

Abstract
Self-tumor antigens that elicit anti-tumor immune/autoimmune responses in responses to interferon-a (IFN-a) stimulation remain poorly defined. We screened a human testis cDNA library with sera from three polycythemia vera (PV) patients who responded to IFN-a and identified a novel antigen, MPD6. MPD6 belongs to the group of cryptic antigens without conventional genomic structure and is encoded by a cryptic open reading frame located in the 3′untranslated region (3′UTR) of myotrophin mRNA. MPD6 elicits IgG antibody responses in a subset of PV patients, as well as patients with chronic myelogenous leukemia and prostate cancer, suggesting that it is broadly immunogenic. The expression of myotrophin-MPD6 transcripts in tumor cells was upregulated in some tumor cells, but only slightly increased in K562 cells in response to IFN-a treatment. By using bicistronic reporter constructs, we showed that the translation of MPD6 was mediated by a novel internal ribosome entry site (IRES) upstream of the MPD6 reading frame. Furthermore, the MPD6-IRES mediated translation, but not myotrophin-MPD6 transcription, was significantly upregulated in response to IFN-a stimulation. These findings demonstrate for the first time that a novel IRES-mediated mechanism is responsible for the translation of unconventional self-antigen MPD6 in responsive to IFN-a stimulation. The eliciting anti-tumor immune response against unconventional antigen MPD6 in patients with myeloproliferative diseases suggests MPD6 as a potential target of novel immunotherapy. Our results also suggest that the enhancement of IRES-mediated translation of unconventional self-antigen(s) by IFN-a is a novel mechanism of IFN-a enhanced anti-tumor immune/autoimmune responses.
Identification and Characterization of T Cell Epitopes Deduced from RGS5, a Novel Broadly Expressed Tumor Antigen.
Authors: Boß Cristina N. , Grünebach Frank , Brauer Katharina M. , Weinschenk Toni , Kanz Lothar , Stevanovic Stefan , Rammensee Hans-Georg , Brossart Peter 
Published: October 2019

Abstract
Identification of tumor-associated antigens (TAA) has resulted in the development of therapeutic vaccines for the treatment of cancer. We applied an integrated functional genomics approach to identify TAA in malignant tissues of patients with renal cell carcinoma (RCC). A comparative DNA chip analysis of tumor and the corresponding non-malignant tissue from patients with RCC followed by sequencing of peptides bound to the HLA-class I molecules by mass spectrometry was applied to identify novel TAA in RCC. To confirm the immunogenicity of identified epitopes cytotoxic T lymphocytes (CTL) were generated using dendritic cells (DC). Utilizing this approach, two peptides derived from RGS5 binding to either HLA-A*02 or 03 were identified. The key function of regulators of G protein-signalling (RGS) is to bind to G protein α subunits and to stimulate their intrinsic GTPase activity. The hydrolysis of guanosine triphosphate (GTP) to guanosine diphosphate (GDP) thereby is accelerated and the inactive heterotrimer more rapidly restored. Thus, RGS proteins inhibit the biological activity of G proteins. Interestingly, it was recently shown that RGS5 is overexpressed in pericytes of newly developing tumor vessels, indicating that RGS5 plays an important role during tumor angiogenesis.
Using RT-PCR analysis we found that RGS5 is expressed on a broad variety of tumor cells including RCC, colorectal, breast and ovarian cancer, malignant melanoma and multiple myeloma as well as in acute and chronic leukemias making this protein an interesting candidate for the development of vaccination strategies to target the tumor cells and the tumor vessels. CTL that were induced using the RGS5 peptides lysed autologous DC pulsed with the cognate peptide or transfected with in vitro transcribed RGS5 RNA as well as HLA-matched tumor cell lines. The specificity and HLA restriction was confirmed using blocking monoclonal antibodies and in cold-target inhibition assays. We next utilized DC transfected with RGS5 RNA to generate specific CTL. Using this approach we confirmed the processing and presentation of the identified peptides by malignant cells. These CTL lysed tumor cells in an antigen specific manner while sparing non-malignant cells. To analyze the induction of RGS5 specific CTL in an autologous setting in patients with malignant diseases, we used blood samples from a patient with acute myeloid leukemia (AML) in complete remission after chemotherapy and were able to generate specific CTL capable of recognizing autologous AML blasts while sparing non-malignant cells.
Our results demonstrate that RGS5 is a novel tumor rejection antigen expressed in a wide range of malignancies that can be applied to target malignant cells and tumor angiogenesis.
Stimulation of Potent Antigen-Specific T-Cell Responses Using Semiallogenic Dendritic Cells Derived from Human Embryonic Stem Cells.
Authors: Su Zhen , Wei Fusheng , Fesperman Susan , Wang Siqing , Dahm Philipp , Vieweg Johannes 
Published: October 2019

Abstract
The major objective of this study is to develop a novel and broadly applicable immunotherapy platform against cancer and infectious diseases. We hypothesized that human embryonic stem cells (hESC) could serve as a source for generating dendritic cells (DC) with potent immunostimulatory function. One advantage of using hESC-derived DC in clinical settings is the ability to generate virtually unlimited amounts of antigen presenting cells for vaccination. Although hESC-derived DC are not genetically identical to the recipient patient, antigen processing and presentation can be facilitated by matching hESC to recipients that share HLA class I alleles. Another advantage of this technology is that hESC express highly polymorphic HLA class II molecules that serve as major rejection antigens, thereby augmenting the antigen-specific T-cell response in the cancer patient.
In the current study, we have established a novel three-step method to differentiate hESC (H9 cell line) into mature DC sequentially through hematopoietic stem cell and myeloid precursor stages. During the first step, 10–15% CD34+ hematopoietic stem cells were generated by co-culturing hESCs with the bone marrow stromal cell line OP9. During the second step, these CD34+ hematopoietic stem cells were differentiated into CD45+CD33+ myeloid precursors and further expanded in the presence of GM-CSF. In the final step, all myeloid precursors were differentiated into mature DC using a cytokine cocktail including GM-CSF, Flt-3L and TNF-α. Using this method, we were able to generate approximately 1 × 108 mature hESC-derived DCs (hESDC) with ≥ 80% purity. These hESDC exhibited a similar phenotype than monocyte-derived DC with high expression of MHC class I, MHC class II, CD11c, CD54, CD40 and the co-stimulatory molecule CD86. Upon activation with proinflammatory cytokines, the hESDC secreted IL-12p70 and migrated in response to MIP-3β. In mixed lymphocyte reaction assays, hESDC exhibited strong allo-stimulatory capacity. Moreover, peptide-loaded mature hESDCs were able to stimulate antigen-specific CD8+ T-cell responses against the EBV peptide BMLF1280-288 and the MART-1 peptide (ELAGIGILTV) in a HLA-A2-restricted manner. Most importantly, hESDC stimulated HLA-A2+ MART-1 peptide-specific CD8+ T cells in vitro that were capable of recognizing and killing the HLA-A2+ melanoma cell line Malena-3M. These data suggest the development of a scalable DC platform that could be applied in clinical immunotherapy protocols.
The Role of Nitric Oxide in Regulation of Red Blood Cell Deformability.
Authors: He Xiaojun , Azarov Ivan , Gordon Beth , Kim-Shapiro Daniel B. , Ballas Samir K. 
Published: October 2019

Abstract
Nitric Oxide (NO) has been suggested to modulate the deformability of red blood cells (RBCs). Bor-Kucukatay (Bor-Kucukatay et al. Am J Physiol Heart Circ Physiol284: H1577, 2003) found that cells incubated with 1 μM of the NO donor sodium nitroprusside lead to a small but significant increase RBC deformability as measured by ektacytometry. However, no significant effect was seen at lower or higher concentrations of sodium nitroprusside or for any concentration of another NO donor, diethylenetriamine NONOate. Kleinbongard (Kleinbongard et al. Blood10; 3992, 2005) found large increases in red cell deformability as a function of added arginine (the substrate for Nitric Oxide Synthase) by measuring the flow rate through filters. On the other hand, using cell aspiration techniques, Bateman (Bateman et al. Am J Physiol Heart Circ Physiol 280; H2848 H2001) found that NO production during sepsis causes a decrease in RBC deformability. Clearly more work is needed to determine the effects of NO on RBC deformability.
The present work was undertaken to further investigate the effect of NO on normal and sickle RBC deformability. ProLi NONOate, arginine, and nitrite (which can be reduced to NO by hemoglobin (Hb), were incubated with blood at various concentrations over a period of 2 hours. Nitrosyl Hb and MetHb formed due to the interaction between NO and RBCs were quantified by electron paramagnetic resonance spectroscopy. The deformability was measured using a flow channel laser diffraction similar to ektacytometry (Huang et al. Am J Hematol67; 151, 2001, Biophys J85; 2374, 2003) with a stress range from 0 to 1,000 Pa. Diffraction patterns produced by deformed cells were analyzed by Matlab®. The deformability coefficients were compared to the control (n=6 per experiment condition). Our results suggested that ProLi NONOate did not significantly effect the deformability of normal RBCs. In a single case, ProLi NONOate improved the deformability of poorly deformable sickle red cells and this result is being studied further. Using our flow channel assay, we did not find any significant affects of arginine on RBC deformability. In addition, our studies involving nitrite, performed under both oxygenated and deoxygenated conditions, suggested that nitrite has no significant effect on RBC deformability. In summary, NO didn’t significantly affect the deformability of normal RBCs, and its potential effects on sickle RBCs needs to be further investigated.
Study of Deferiprone or Deferoxamine Versus Combination Therapy in Iron-Loaded Thalassemia Patients in Taiwan.
Authors: Peng Ching-Tien , Wu Kang-Hsi , Tsai Chang-Hai , Yang Chao-Ping , Wang Lin-Yen , Jang Ren-Chin , Hsiao Chih-Cheng 
Published: October 2019

Abstract
BACKGROUND: Deferiprone (L1) has been suggested to be an effective oral chelation therapy for regular transfusion thalassemia major patients. The purpose of this study was to test the efficacy either of L1 or DFO alone, or combination use, and also monitor the safety of L1 in Taiwan.
MATERIALS AND METHODS: From April 1999 to December 2005, 114 thalassemic patients from 5 treatment centers were enrolled in this program. L1 at the standard dose of 75mg/kg was given to 57 patients. The mean administered dose of L1 was 72.5mg/kg body weight divided into three doses per day. DFO at the standard dose of 30–50mg/kg/day at least 5 days/week was given to 26 patients, and the mean DFO dose was 46.5mg/kg/day 5 or more days/week. Combined therapy of daily L1 with subcutaneous DFO was given 2 to 6 days each week to the other 31 patients. The mean administered dose of DFO was 45.7mg/kg body weight 2 to 6 days per week, and the mean L1 dose was 71.5mg/kg/day divided into 3 doses per day. The therapeutic efficacy and potential side effects on cardiac and/or hepatic systems of these patients were assessed by left ventricular ejection fraction, T2-weighted magnetic resonance imaging (T2-MRI) and biochemical parameters and liver biopsies.
RESULTS: No significant liver function impairment, neutropenia or arthropathy ever occurred. Only one patient was found to have transient leukopenia (he had a coincidental viral infection at that time). Three patients had temporary G-I upset but no one required discontinuation of L1. The serum ferritin level was significantly lower in 9 patients receiving L1 for more than 6 years (p=0.04), 22 patients receiving L1 for 2–3 years (p<0.01) and 31 receiving the combined therapy (p=0.01), yet significantly higher in those receiving DFO only (p<0.01). The combined therapy group witnessed a more significant ferritin decrease than the L1 alone or DFO alone groups and hepatic function improved or stabilized notably. Besides a decrease of iron, no marked pathohistological changes were observed in the liver biopsies. GPT had a decreasing trend in 3 subgroups of L1 & combination therapy group, however there was no statistic significance. Cardiac studies showed a marked recovery of signal intensity in the heart T2-MRI and increased LVEF, indicating a significant reduction of iron load in the heart in L1 use and in over 50 -month combination therapy groups
CONCLUSIONS: We conclude that in this large group of thalassemia major patients, L1 was a safe long-term method of iron chelation. Combining subcutaneous DFO with L1 therapy is an improved method not only for patients inadequately chelated but also by lowering the DFO dosage to less than 25%. This study also shows that the combined therapy is associated with an improvement in heart function.
Splenic Infarction Due to Concomitant Hereditary Spherocytosis and Sickle Cell Trait.
Authors: Yates Robin , Buchanan George R. , Ginsburg Howard , Fefferman Nancy , Greco M. Alba , Borys Dariusz , Blei Francine 
Published: October 2019

Abstract
The occurrence of hereditary spherocytosis (HS) and sickle cell trait (HbAS) in the same patient is rare, with just 21 cases reported in the literature. Although six of these cases had splenic infarction or sequestration, none were reported as having a precipitating event. However, in persons with HbAS, splenic sequestration or infarction has been reported following exposure to high altitudes, including pressurized commercial aircraft. The interaction of these 2 (HS and HbAS) disorders can be explained by the hypoxic and acidic environment in the spleen. This environment causes spherocytes containing HbAS to be both less deformable and more prone to sickling, which promotes sluggish blood flow within the spleen and leads to sequestration and infarction. We have encountered a 10-year-old Caucasian girl with both HS and HbAS who experienced left upper quadrant pain during and following a flight on a commercial aircraft. Past medical history included episodes, although less severe, of left upper quadrant pain after traveling to high altitudes on automobile trips or on commercial aircraft. Family history included the dual diagnoses of HbAS and HS in the patient’s mother, for which she underwent splenectomy as a teenager, and a diagnosis of HS in her 9-year-old brother. At presentation, she had significant left upper quadrant tenderness upon palpation that was associated with voluntary guarding. There was no rebound or involuntary guarding. The spleen was noted to span 3–4cm below the left costal margin. Laboratory values included a hemoglobin of 10.8 (g/dl), reticulocyte count of 7.5%, total bilirubin of 3.0 (mg/dl), and lactate dehydrogenase of 1160 (U/L), all of which were similar to her baseline values. Abdominal ultrasound and cat scan demonstrated infarction in the inferior pole of her spleen. Despite supportive medical treatment, she required immediate splenectomy to relieve her symptoms and prevent recurrence. Her operative and post-operative courses were uneventful. This first report of a patient with both HS and HbAS who suffered splenic infarction after traveling on a commercial aircraft demonstrates how two different inherited red blood cell abnormalities can interact and lead to clinically significant complications.
Hydroxyurea-Mediated Reactivation of Fetal Genes in Primary Erythroid Cell Cultures from Beta-Thalassemia Patients.
Authors: Calzolari Roberta , Maggio Aurelio , Pecoraro Alice , Borruso Vito , Troia Antonio , Acuto Santina , D’Apolito Danilo , Ferro Leda , Di Marzo Rosalba 
Published: October 2019

Abstract
Fetal hemoglobin (HbF) is frequently increased in the hemoglobinopathies such as sickle cell anemia and b-thalassemia. Epidemiological studies have indicated that an increase in HbF ameliorates the clinical symptoms of these diseases (Rodgers and Rachmilewitz - Bri J. Haemat. 91: 263 -1995).
In sickle cell anemia, HbF containing red blood cells have a lower concentration of sickle hemoglobin (HbS), and the HbF itself inhibits HbS polymerization, decreasing cell sickling process (Eaton and Hofrichter - Science 268:1142 -1995). In b-thalassemia patients, HbF partially compensate HbA deficiency and could potentially improve RBC survival resulting in an increase of hemoglobin levels.
Hydroxyurea (HU) is one of the pharmacological agents currently used to stimulate HbF synthesis in patients with sickle cell anemia and more recently has been tested in clinical trials for b-thalassemia patients too (Olivieri et al. Ann. NY Acad Sci850:100-1998; Rigano et al. Hemogl.21(3): 219- 1997; Dixit et al. Ann. Haematol. 84: 441 -2005).
The mechanism involved in the HU-mediated changes is still unclear. It may involve a selection of a minor pre-existing subpopulation of F-cells that has a growth and/or survival advantage (cellular mechanism). This mechanism may be particularly effective for cells derived from patients with hemoglobinopathies, where F-cells may be resistant to “ineffective erythropoiesis”. An alternative mechanism could involve stimulation of HbF in all or the majority of cell-population by direct induction of g genes (molecular mechanism).
Here we report the analysis on thalassemia patients homozygoutes for Lepore genotype that present high levels of fetal hemoglobin.
We combined the use of primary erythroid cell culture from peripheral blood stem cells of these patients, with primary transcript in situ hybridization (RNA-FISH) of the g and b globin genes to investigate the mechanism of action of hydroxyurea in adult erythroid cells.
RNA-FISH on erythroid cell cultures from these patients reveals that the majority of cells express one g allele only (g: 75.2 %, g:g 19.6%). The analysis in hydroxyurea-treated cultures shows the increase of cells transcribing both g-alleles, indicating the reactivation of fetal genes (g: 58.1%, g:g 40%).
This evidence suggests that the molecular mechanism is involved directly on fetal genes reactivation to increase fetal hemoglobin production in HU-treated patients.