bluuebunny/crossref_metadata_2025_split huggingface dataset:
Primary Prophylaxis Prevents the Onset of Arthropathy: Longitudinal Results of 603 Boys with Severe Factor VIII Deficiency Analyzed through the Centers for Disease Control Universal Data Collection (CDC UDC) Study.
Authors: Manco-Johnson Marilyn J. , Wang Chengbin , Konkle Barbara A. , Ingram-Rich Robina , Humes Steven , Soucie J. Michael 
Published: October 2019

Abstract
BACKGROUND: Arthropathy is the most common complication of severe factor VIII deficiency and causes chronic pain, functional disability and increased cost of care. Early institution of primary prophylaxis (PP) was found to prevent hemophilic arthropathy at age 6 years in a prospective randomized clinical trial including 65 boys (NEJM2007;357:535–44). The current study was conducted to determine the prevalence and efficacy of primary prophylaxis to prevent joint disease in the larger population of young children enrolled in the CDC UDC data base.
METHODS: Eligibility for this analytic project included boys with factor VIII < 1% entered onto the UDC study at or before 36 months of age. Treatment was clinically determined and defined as prophylactic which could be primary (PP, initiated at or before 36 months), secondary (SP, initiated at or following 42 months), or indeterminate (IP, initiated between 36 and 42 months); or episodic (E, without routine prophylaxis). A target joint was defined as one with four or more bleeding events within 6 months. Kaplan-Meier curves were used to evaluate survival without target joint development over time; Cox modeling was employed to assess risk factors for target joint development. Univariate and multivariable analyses were used to model risk factors for bleeding.
RESULTS: Of 603 eligible boys, treatment included: PP in 252 (42%), SP in 73 (12%), E in 240 (40%). 38 boys (6%) with IP were excluded from the analysis. In comparison to children treated with E, those on PP were, on average: older (2.17 vs 1.87 years, p < 0.0001); more likely to have had a bleeding event (96 vs 91%, p=0.002); more likely to have received home infusions (90 vs 49%, p < 0.0001); and more likely to have suffered an intracranial hemorrhage (11.3 vs 4.1%, p=0.02). Despite their increased experience with joint and other bleeding events, during the period of prospective observation, children on PP had significantly less joint bleeding (OR 0.51, CI 0.39–0.66, p < 0.0001) and showed improved survival time without onset of a target joint (p = 0.002 overall, and 0.004 between PP and E, figure 1). Results for muscle or any bleeding were similar. In multivariable analysis PP exerted the greatest protective effect on risk of target joint development (hazard ratio 0.38, 95% CI 0.22–0.66, P = 0.001). Factors associated with E therapy included non-white race (p < 0.0001) and insurance coverage other than commercial (p = 0.004), although these factors were not associated with poor joint outcomes.
CONCLUSION: In a large population of children with severe hemophilia A on the CDC UDC registry, PP prevents joint and other bleeding and delays target joint development. Factors limiting application of PP must be explored and addressed.
Figure Figure
Alteration of Circulating Regulatory T Cells in Japanese Adult Patients with Immune Thrombocytopenic Purpura after Eradication of Helicobacter Pylori.
Authors: Miyazaki Koji , Danbara Mikio , Ohsaka Manabu , Suzuki Yuhko , Horie Ryouichi , Higashihara Masaaki 
Published: October 2019

Abstract
Immune thrombocytopenic purpura (ITP) is an autoimmune disorder, and eradication of Helicobacter pylori (HP) has been demonstrated to be an effective and tolerable firstline treatment for the patients infected with HP in Japan. However the mechanism has still remained to be uncovered. Recently CD4+CD25high+Foxp3+ regulatory T cells (Tregs), which regulate autoreactive T cells, have been identified, and suggested to play an important role in pathogenesis of ITP, as well as other autoimmune disorders. It has been shown that Tregs are reduced and suppressed in the ITP patients. And a recent report has demonstrated that the defective Tregs are restored upon the rituximab treatment. However, the effects of HP eradication therapy on Tregs have not been determined. The aim of this study is to investigate the circulating Tregs in ITP patients treated with HP eradication. And we also attempted to elucidate the mechanisms of the treatment. The peripheral blood CD4+CD25high+ Tregs were measured by flow cytometry before and after the treatment in 21 Japanese adults with HP-positive chronic ITP. We also confirmed the expression of Foxp3 in this cellular population with the permeabilized mononuclear cells. Among 21 patients, the platelet counts increased in 13 cases (responders), but not in 9 cases (non-responders). In responders the numbers of Tregs have been restored, but not in non-responders after the treatment. It is interesting here that the amounts of Tregs were still transiently elevated in an early phase (2–3weeks) after the treatment in some non-responders without recovery of the platelet counts. Furthermore, in three cases, who failed in pylorus elimination, Tregs were also transiently increased in number associated with brief recovery of the platelet counts, and reduced to the initial level in about two months. After the successful re-eradication, the numbers of Tregs and platelets have been restored. From these results it is shown that HP eradication can modulate Tregs to increase the platelet counts for ITP patients. We also demonstrate that the increase of Tregs by HP eradication was more rapid than that by rituximab, which required about three months. Further, it might be suggested that there are two phases of the therapy with HP eradication for ITP. In an initial stage, the therapy itself could have an effect modulating the immune systems to potentiate Tregs. Some drugs, such as macrolide antibiotics including clarithromycin have been demonstrated to be a potent immunomodulator. However, this phase is not sufficient for the successful treatment, as shown in the cases of failure in HP elimination. In a retentive stage, extermination of HP is also necessary for sustainment of restored Tregs.
The Uroporphomethene Inhibitor Causitive for Porphyria Cutanea Tarda (PCT) Is Generated by Oxidation of Hydroxymethylbilane (HMB)
Authors: Phillips John D , Bergonia Hector A , Reilly Christopher A , Franklin Michael R , Kushner James P 
Published: October 2019

Abstract
Porphyria cutanea tarda (PCT) is caused by a uroporphomethene (UPN) inhibitor of uroporphyrinogen decarboxylase (Phillips, JD et al, Proc Natl Acad Sci U S A. 2007 104(12):5079–84). In the absence of the inhibitor very little uroporphyrin (URO) is secreted in the urine and the excreted URO is predominantly the isomer-III (URO-III). In contrast, in patients with PCT and mice with experimental PCT the predominant porphyrin excreted in urine is uroporphyrin-I (URO-I). Direct measurements of uroporphyrinogen III synthase activity (UIIIS) in mice with PCT have shown no reduction in activity indicating that an alternate mechanism is responsible. We hypothesized that oxidation of a single bridge carbon of HMB, the product of the porphobilinogen deaminase reaction, is the initial event leading to the porphomethene. Our structural data indicate that oxidized HMB cannot serve as a substrate for UIIIS (Schubert, HL et al, Biochem J. 2008 47(33):8648–55) but can non-enzymatically cyclize to form UPN I. UPN I would then serve as the inhibitor of uroporphyrinogen decarboxylase but and then be fully oxidized to yield URO-I. To test this hypothesis we generated authentic UPN’s I and III. URO’s I and III were reduced with palladium on carbon to yield uroporphyrinogens (Uro’gen) I and III. UPN’s I and III were then generated by controlled photo-oxidation of the corresponding Uro’gens. We then utilized LC-MS to separate and characterize the synthesized UPN’s. Retention times of the two compounds differed by approximately 1 min (UPN III elutes prior to UPN I) but the profile of the daughter ions derived from the UPN’s (molecular mass 835 Da) was identical. The same approach was applied to clarified liver cytosol from mice with experimental PCT. The retention time of the 835 Da UPN identified in mouse liver cytosol corresponded to that of the synthesized UPN I and yielded identical daughter ions. These data support the hypothesis that oxidation of HMB leads to formation of UPN I in vivo. This is concordant with our view that UPN I serves both as the inhibitor of URO-D and as the source of the excess URO-I found in PCT.
Human SIRPA Polymorphism Modulates Macrophage-Mediated Suppression of Human Hematopoiesis
Authors: Kuriyama Takuro , Takenaka Katsuto , Kumano Takashi , Oku Seido , Urata Shingo , Nagafuji Koji , Dick John E. , Danska Jayne S , Akashi Koichi 
Published: October 2019

Abstract
Signal regulatory protein-α (SIRPA) is an immunoglobulin superfamily transmembrane protein with intracellular docking sites for two Src homology domain containing tyrosine phosphatases, and most abundantly expressed in neurons and myeloid cells. SIRPA is a critical immune inhibitory receptor on macrophages. CD47 is a ligand for SIRPA, and CD47 interaction with SIRPA serves as a ‘self-recognition’ that prevents phagocytosis of the cells expressing CD47. Recently, we (Nature immunology 2007) identified polymorphism in murine Sirpa as a critical modulator of engraftment in xenogeneic model of human-to-mouse hematopoietic stem cell transplantation, and non-obese diabetic (NOD) mouse Sirpa polymorphism has far greater reactivity with human CD47 than that of the respective alleles of other strains resulting in effective engraftment of human hematopoietic cells. In addition, as well as the mouse, human SIRPA is highly polymorphic in the IgV domain. Then, we examined whether polymorphism in SIRPA induced the difference for suppressive effect of human macrophage on hematopoiesis. First, we examined sequence alignment of SIRPA IgV domain (exon 3 of SIRPA) of healthy control 18 people. We identified two major variants; 6 people with variant1 (V1) and 12 people with variant2 (V2). The SIRPA amino acid sequences of V1 and V2 are different in 13 residues, and its residues in orthologous position between species that is polymorphic between NOD and other strains as well as between V2 and V1. Next, we examined that whether there is a difference in the effect on hematopoiesis between V1 macrophage and V2 macrophage by long term culture-initiating cells (LTC-IC) assay on MS-5 stromal cells. For macrophage preparation, peripheral mononuclear cells from healthy donors were purified by positive selection using MACS CD14 Micro Beads (Miltenyi Biotec), and they were incubated with M-CSF for 3 days. For LTC-IC assay, 5×102 differentiated macrophages were seeded onto established MS-5 stroma in 96-well tissue culture plates. The next day, human hematopoietic CD34+ cells were seeded at doses of 102 to 5×103 cells per well and cultured for 4 weeks. At the end of the culture, cells were detached and plated into methylcellulose progenitor assays. Macrophage had a suppressive effect on the number of LTC-IC in all cultures. There was tendency that V1 macrophage had a greater suppression compared to V2 macrophage, however the differences between V1 and V2 were marginal (p=0.03). These data suggests that human macrophages have suppressive effect on hematopoiesis, and human SIRPA polymorphism modulates macrophage-mediated suppression of hematopoiesis in allogenic model, likewise in xenogeneic model of human-to-mouse hematopoietic stem cell transplantation. Moreover, SIRPA polymorphism might be related to graft failure in the allogenic hematopoietic stem cell transplantation.
The Potent Deacetylase Inhibitor Trichostatin a (TSA) Increases CXCR4 Expression in Hematopoietic Stem/Progenitor Cells by Chromatin Remodelling
Authors: Gul Hilal , Marquez-Curtis Leah A. , Lo Jennifer , Jahroudi Nadia , Turner A. Robert , Larratt Loree M. , Janowska-Wieczorek Anna 
Published: October 2019

Abstract
Stromal-cell derived factor (SDF)-1α/CXCL12 and its cognate receptor, CXCR4, play a crucial role in the trafficking of normal hematopoietic stem/progenitor cells (HSPC) and their homing/retention in bone marrow. Consequently, modulation of CXCR4 expression in HSPC could be applied therapeutically to improve the efficiency of HSPC transplantation. It is known that gene expression can be regulated by chromatin remodelling. Two groups of histone modifying enzymes, histone acetyltransferase (HAT) and histone deacetylase (HDAC) participate in the regulation of chromatin structure, and hence gene expression. Disruption of normal HAT or HDAC activities has been found in many human cancers. Recently, several structurally diverse and highly specific HDAC inhibitors (HDI) have been reported. They act as strong modulators of growth, differentiation and apoptosis in several types of cancer, particularly acute myeloid leukemia (AML). However, very little is known regarding the effects of HDI on HSPC. We have previously shown that a specific short-chain fatty acid HDI, valproic acid (VPA), enhances CXCR4 expression and function in normal HSPC (Blood2007: 110; 425a). In order to determine whether other structurally diverse classes of HDI are able to influence CXCR4 expression in HSPC through chromatin remodelling, we investigated the effect of potent hydroxamic acid HDI Trichostatin A (TSA) on CXCR4 in normal HSPC. We examined the effect of TSA on CXCR4 expression (by FACS and real-time RT-PCR), modulation of CXCR4 transcription (chromatin immunoprecipitation (X-ChIP) analysis) and on functional response towards an SDF-1α gradient (by chemotaxis assay) of HSPC (CD34+ cells from cord blood (CB) and the models of immature hematopoietic cells expressing CD34 antigen, namely AML cell lines KG-1a and KG-1). Cells were incubated for 24 h in IMDM supplemented with 20% FCS in the presence of TSA (0.1 μM). We found that TSA increases the percentage of CXCR4-expressing CB CD34+, KG-1a, KG-1 cells (2.5-, 8- and 3-fold, respectively). This effect was also confirmed at the mRNA level in CB CD34+, KG-1a and KG-1 cells (by about 2.5-, 5- and 2.5-fold up-regulation, respectively). Moreover, X-ChIP analysis showed a significant increase in association of acetyl-histone H4 binding to the CXCR4 promoter in CB CD34+ and KG-1 cells (2- and 1.7-fold, respectively). TSA was also shown to significantly increase the chemotaxis of KG-1a cells towards SDF-1α (20 ng/mL), which was inhibited by AMD3100, a potent antagonist of CXCR4. We conclude that other HDI such as TSA regulate CXCR4 expression in HSPC by chromatin remodelling and we suggest that priming of HSPC with HDI may improve their homing and engraftment into bone marrow, especially in CB transplantation.
Induction of Invariant NKT Cell-Dependent Alloreactivity by Administration of G-CSF after Bone Marrow Transplantation
Authors: Morris Edward S , MacDonald Kelli P A , Kuns Rachel D , Morris Helen M , Banovic Tatjana , Don Alistair LJ , Rowe Vanessa , Burman Angela C , Markey Kate A , Godfrey Dale I , Smyth Mark J , Hill Geoff 
Published: October 2019

Abstract
G-CSF is often used to hasten neutrophil recovery following allogeneic bone marrow transplantation (BMT), but the clinical and immunological consequences invoked remain unclear. We examined this in murine models and found that administration of both standard G-CSF and pegylated G-CSF early after BMT significantly increased graft-versus-host disease (GVHD). This effect was seen in the B6 → B6D2F1, BALB/c → B6 and C3H.SW → B6 systems of GVHD to either MHC or multiple minor histocompatibility antigens. This effect was dependent on total body irradiation (TBI) rendering host dendritic cells (DC) responsive to G-CSF by up-regulating their expression of the G-CSF receptor as determined by real-time PCR. This induction of G-CSFR expression was not seen following busulfan (Bu), cyclophosphamide (Cy) or fludarabine. The enhanced GVHD was present when G-CSF was administered to both WT and G-CSFR−/− donors but not G-CSFR−/− recipients, confirming that host signalling was critical for this effect. G-CSF administration after BMT had no effect on inflammatory cytokine generation but enhanced in vivo CTL activity after BMT when administered to WT but not G-CSFR−/−, CD1d−/−, IFNgR−/− or CD40−/− recipients. Furthermore, donor iNKT cell activation was absent in CD11c Diptheria Toxin Receptor recipient transgenic mice depleted of dendritic cells (DC) by diphtheria toxin and treated with G-CSF. Thus, stimulation of host DC by G-CSF subsequently unleashed a cascade of events characterized by CD1d dependent donor iNKT cell activation, IFNg secretion and CD40-dependent amplification of donor CTL function during the effector phase of GVHD. Critically, the detrimental effects of G-CSF on GVHD were present when administered early following TBI conditioning and at a time when residual host APC were still present (day +1), but had no effect when administered at day +8 when host DC were not detectable by phenotypic or functional analysis. This is consistent with the inefficient cross presentation of host Ag within MHC class I by donor DC after BMT. In addition, the administration of G-CSF after Bu/Cy conditioning had no effect, perhaps explaining the conflicting and somewhat controversial clinical studies from the large European and North American BMT registries since TBI conditioning predominated only in the positive European study. These data have major implications for the use of G-CSF in disease states where NKT cell activation may have important effects on outcome and suggest a guide to the safe use of G-CSF after allogeneic BMT.
Soluble Lymphotoxin Plays a Critical Role in Acute Graft-Versus-Host Disease
Authors: Markey Kate A , Burman Angela C , Kuns Rachel D , Morris Edward S , Banovic Tatjana , Raffelt Neil C , Olver Stuart D , Engwerda Christian R , Korner Heinrich , MacDonald Kelli P A , Hill Geoff R 
Published: October 2019

Abstract
TNF is a key cytokine in the effector phase of both graft-versus-host disease (GVHD) and the graft-versus leukemia (GVL) effect after bone marrow transplantation (BMT). TNF neutralizing antibodies are now established as effective therapeutic adjuncts for the treatment of severe acute GVHD. TNF signals through the p55 and p75 TNF receptors (TNFR), which are also receptors for the soluble lymphotoxin homotrimer (LTα3). The membrane-bound lymphotoxin heterotrimeric molecule (LTα1β2) signals through the LTβ receptor. The function of these molecules in GVHD remains unknown. Pharmacological agents are available which block either TNF alone, or both TNF and LTα3 and elucidating the roles of these molecules in GVHD is essential for the design of rational therapeutic strategies.
We have employed multiple well established preclinical models of GVHD to investigate the roles of these molecules. We first used a functional bioassay to confirm that a human TNFR:Fc construct was capable of preventing physiological activity of both recombinant murine TNF and LTα3. Irradiated B6D2F1 recipients were then transplanted with allogeneic B6.TNF−/− bone marrow and T cells, then treated with control Ig or the TNFR:Fc. Suprisingly, the TNFR:Fc provided significant protection from GVHD mortality in the absence of donor derived TNF (median survival 25.5 vs 35 days, P<0.0002). Having observed a potentially TNF-independent but TNFR dependent pathway in the generation of GVHD we next examined mRNA expression by real-time PCR. This demonstrated that the majority of LTα mRNA was within donor CD4 T cells and that expression was unbalanced (> 3 logs more LTα than LTβ). We subsequently developed the first reported ELISA to assess murine LTα3 protein and were able to clearly demonstrate LTα3 production by wild-type (WT) CD4 T cells seven days after BMT. To further examine this molecule, we performed experiments using WT, LTα−/− or LTβ−/− donor grafts in two BMT systems (B6 → B6D2F1 and B6 → BALB/c). Recipients of LTα−/− grafts were protected from GVHD (P<0.01) in both systems while LTβ−/− grafts induced GVHD that was equivalent to, or more severe than in recipients of WT grafts. This data confirms soluble LTα3 but not membrane bound LTα1β2 as an additional pathogenic molecule in GVHD. Using a luciferase-expressing, host-type leukemia (P815) in the B6 → B6D2F1 system, relapse occurred at an identical rate in recipients of WT and LTα−/− grafts confirming that GVL is maintained in the absence of donor derived soluble LTα3. Thus the combined neutralization of both LTα3 and TNF represents an important and logical therapeutic intervention in transplant medicine.
Mechanisms of Synergy Between Toll-Like Receptor 4 and Triggering Receptor Expressed on Myeloid Cells-1 in Human Neutrophils
Authors: Radsak Markus P , Schild Hansjörg , Haselmayer Philipp , Salih Helmut R 
Published: October 2019

Abstract
The triggering receptor expressed on myeloid cells 1 (TREM-1) is an important player in the innate inflammatory response to microbial infections. Activation and expression of TREM-1 by polymorphonuclear neutrophils (PMN) occurs in concert with Toll-like receptors (TLR) such as TLR4 for bacterial lipopolysaccharide. However, it is currently unclear how this is mediated on a molecular level. Using pharmacologic inhibitors and western blot analysis we demonstrate that phosphatidyl inositide 3-kinase, phospholipase C and the mitogen activated kinase p38 are essential for the TREM-1 and TLR4 mediated respiratory burst of human PMN. The down stream phosphorylation of protein kinase B and extracellular signal related kinase show characteristic phosphorylation patterns upon single or co-ligation indicating individual activation pathways of both receptors. TREM-1, but not TLR4 mediated respiratory burst depended on calcium flow via store operated calcium entry channels, while transient receptor potential channels were important for TLR and TREM-1. Taken together, we provide new insights on the mechanisms how TREM-1 and TLR interact creating synergistic activation in PMN. These results shed a new light on our understanding how the innate inflammatory responses are regulated and might contribute to the development of future concepts for the treatment of severe inflammatory conditions such as sepsis.