bluuebunny/crossref_metadata_2025_split huggingface dataset:
Desulfated Heparin Blocks Platelet Activation Induced by HIT Antibodies / Heparin: A New Approach to Patient Management.
Authors: Prechel Margaret , Jeske Walter P. , McDonald Meredith K. , Drenth Amanda F. , Walenga Jeanine M. 
Published: April 2020

Abstract
Heparin-induced thrombocytopenia (HIT) represents a disease spectrum triggered by an immune response to heparin. The most dramatic clinical expression of HIT is HIT antibody-driven thrombosis. Direct thrombin inhibitors (DTIs) are a promising new class of drugs for treatment of the acute phase of HIT; however, they have a narrow safety/efficacy window (high bleeding risk) and morbidity/mortality have not been eliminated. In addition, the high probability of developing thrombosis in HIT combined with extreme mortality, has led to a bias for prophylactic treatment. Thus, there remains a clinical need to identify optimal treatment options for patients with HIT. A new concept is to use an agent that can effectively compete with heparin in a manner that prevents the HIT antibody from inducing platelet activation, i.e., amelioration. We evaluated a 2-O, 3-O desulfated heparin (ParinGenix, Inc.; Tucson, AZ) to determine its ability to ameliorate HIT antibody/heparin induced platelet activation. The test agent was added to a mixture of known-reactive platelets and pre-formed immune complexes (heparin, PF4, HIT antibodies), and SRA and flow cytometry were performed. Due to the inherent biological variability of HIT antibodies, sera from four different patients (clinically diagnosed as HIT; SRA positive) were used. Two concentrations of heparin which reflect typical prevention (0.1 U/ml) and treatment (0.5 U/ml) clinical doses were used. The 2-O, 3-O desulfated heparin produced an amelioration of HIT antibody/heparin induced platelet activation as demonstrated in the SRA by inhibition of 14C serotonin release from activated platelets, and in flow cytometric analysis by inhibition of platelet microparticle formation and platelet cell surface P-selectin expression. Significant amelioration activity was initiated at 6.25 μg/ml 2-O, 3-O desulfated heparin and complete inhibition of the induced platelet activation (equal to the 100 U/ml heparin HIT assay ‘no response’) was achieved with 50 μg/ml of the agent. Since this new treatment approach blocks platelet activation caused by HIT antibody/heparin (not a characteristic of DTIs), we propose that a non-anticoagulant glycosaminoglycan (GAG) may be useful in improving the clinical management of patients with HIT. The concept of amelioration differs from all previous options for the clinical management of patients with HIT including the use of danaparoid, fondaparinux, DTIs and other drugs that target thrombin/thrombin generation inhibition. Although not directed at platelet activation inhibition, this type of GAG effects an inhibition of HIT antibody mediated platelet activation, which is the source of the pathophysiology of HIT. The data of this study suggest that this 2-O, 3-O desulfated heparin may be effective as either an adjunct or sole treatment of an ongoing HIT pathology, or as a preventive measure in patients who will be exposed to heparin.
A Novel Frameshift Mutation in a Case of Familial May-Hegglin Anomaly.
Authors: Cutler Jacky A. , Mitchell Mike J. , Savidge Geoffrey F. 
Published: April 2020

Abstract
The May-Hegglin anomaly is a rare, autosomal dominant condition that is characterised by thrombocytopaenia, anisocytosis, and Dohlë-like inclusion bodies in leukocytes. The disorder generally results in a mild bleeding tendency. May-Hegglin anomaly results from defects in the MYH9 gene, which encodes the heavy chain of nonmuscle myosin IIA. The gene maps to chromosome 22q12.3-q13.2, comprises 40 exons, and produces a 5.8kb mRNA transcript.
We present a case of familial May-Hegglin anomaly. This kindred was first identified, and published, in 1964; but no further analyses were carried out until recently.
The proband, now aged 80 years, has a life long history of easy bruising and epistaxis, although the symptoms have improved with increasing age. Her sister (deceased) and daughter reported similar symptoms: the daughter requiring splenectomy for thrombocytopaenia at age 9 years. The proband has also suffered from deafness for in excess of 60 years.
Recent investigations of the proband revealed a mild thrombocytopaenia, and the characteristic blood film of May -Hegglin anomaly. Platelet and neutrophil morphology were also consistent with the disorder. Karyotype analysis revealed good quality G banding throughout chromosome 22, and no gross alteration in the region 22q11.2., 22q13. Direct sequencing of the MYH9 gene identified a heterozygous deletion of a single base in a run of Gs at nucleotide positions 5818–5821 (codons 1940–1941). This frame shift mutation leads to the creation of a premature termination sequence at codon 1947, and the loss of the last fourteen amino acids. The deletion introduces a restriction site, and endonuclease digestion of a panel of normals failed to detect this change. Together with the fact that this is a conserved region across species (human, rat, rabbit, chicken), it is probable that this deletion is the causative mutation in this kindred.
This novel mutation represents the second reported defect affecting the tail-piece domain in May-Hegglin anomaly. The protein’s two heavy chains homodimerise in this region to form a tail, and multiple polypeptides associate through this tail domain to form bipolar filaments. Therefore, a mutation in this domain may exert its effect by the inhibition of myosin filament assembly.
To date, approximately 20 different mutations have been described in the MYH9 gene giving rise to not only May-Hegglin anomaly, but also the similar disorders of Fechtner syndrome, Sebastian syndrome and Epstein syndrome. In fact some mutations have been identified in patients with different diagnoses. The proband presented here has a long standing diagnosis of May-Hegglin anomaly, but does exhibit one symptom (deafness) more usually associated with Fechtner syndrome. The genetic, phenotypic and clinical overlap in these disorders suggest that these actually represent allelic variations of a single genetic disorder, which should more correctly be termed ‘MYHIIA syndrome’.
Hepatocyte Growth Factor Ameliorates Chronic Graft-Versus-Host Disease.
Authors: Imado Takehito , Iwasaki Tsuyoshi , Kuroiwa Takanori , Okamoto Takahiro , Hara Hiroshi , Sano Hajime 
Published: April 2020

Abstract
Since chronic graft-versus-host disease (GVHD) is a major cause of morbidity and mortality in allogeneic hematopoietic stem cell transplantation (HSCT), a new strategy to prevent chronic GVHD is necessary. We previously showed that hepatocyte growth factor (HGF) protected against lethal effects of acute GVHD and promoted T cell reconstitution in murine models (J Clin Invest107;1365:2001, Blood 2004 in press). We have now investigated whether HGF can protect against chronic GVHD. (C57BL/6 x DBA/2) F1 (BDF1) recipient mice were lethally irradiated (9 Gy) and co-transplanted with 5 x 106 bone marrow cells and 1.5 x 107 spleen cells from C57BL/6 donor mice. At the same time, mice were injected intramuscularly with 8 microgram of the human HGF gene, or with a control mock vector, every week using liposomes containing the hemagglutinating virus of Japan. Untreated GVHD recipients showed significant atrophy of the spleen and the thymus on day 56 after transplantation, while HGF treatment effectively prevented this atrophy. Splenic T cells from untreated GVHD recipients proliferated and produced significant amounts of IL-4 after being exposed to irradiated BDF1 spleen cells in vitro, while those from HGF-treated GVHD recipients did not. These results suggest that host-reactive type 2 T helper (Th2) cells were eliminated in the HGF-treated GVHD recipients by protection against thymic damage caused by acute GVHD, thereby preventing the progression to chronic GVHD. We next used DBA/2 into BDF1 models to determine the effect of HGF on another type of chronic GVHD. Chronic GVHD was induced by injection of DBA/2 spleen cells (9 x 107) into non-irradiated BDF1 mice. HGF treatment effectively prevented proteinuria and histological changes associated with glomerulonephritis. Liver sections of untreated GVHD recipients revealed prominent primary biliary cirrhosis (PBC)-like changes, as indicated by lymphocyte infiltration into the periportal area of the liver. HGF treatment effectively prevented these histological changes. HGF treatment greatly reduced the number of splenic B cells and the serum IgG and anti-DNA antibody levels. Furthermore, HGF treated GVHD recipients showed down-regulation of IL-4 mRNA expression in the spleen, liver and kidneys. IL-4 induction from DBA/2 CD4+ T cells stimulated by irradiated BDF1 CD11c+ dendritic cells (DCs) was significantly inhibited by addition of recombinant HGF (10 ng/ml) in vitro. Taken together, these results indicate that HGF can prevent chronic GVHD by eliminating host-reactive Th2 cells in the thymus, or by inhibition of Th2 cell induction by interaction with DCs.
Perforin and Fas Ligand Are Required for the Regulation of Alloreactive CD8+ T Cells.
Authors: Maeda Yoshinobu , Levy Robert B. , Reddy Pavan , Liu Chen , Teshima Takanori , Ferrara James L.M. 
Published: April 2020

Abstract
We evaluated the role of Fas ligand and perforin, the major T cell-mediated cytotoxic pathways that regulate T cell homeostasis, in a CD8+ T cell mediated model of graft-versus-host disease (GVHD) where donor and recipients differ at a single MHC class I antigen (B6 → bm1). Lethally irradiated (11Gy) bm1 mice were transplanted with T cell depleted BM and CD8+ T cells from either wild type (wt) or cytotoxic double deficient (cdd, deficient in both pathways) B6 donors. We hypothesized that cdd CD8+ T cells would be unable to mediate significant GVHD. Contrary to our hypothesis, recipients of cdd donor CD8+ T cells demonstrated significantly greater histopathologic damage from GVHD and increased serum levels of IFN-gamma and TNF-alpha compared to controls (Table 1). In order to understand this increase, we evaluated the in vivo expansion of donor T cells in these recipients as well as in BMT recipients of allogeneic CD8+ T cells from FasL deficient (gld) and perforin deficient (pfp−/−) donors. CD8+ wt T cells expanded until at day 10 after BMT, followed by a rapid decline. By contrast, cdd CD8+ T cells expanded continuously up to day 30 after BMT, peaking at almost one hundred times the number of wt T cells. gld T cells showed kinetics similar to wt T cells, whereas the pfp−/− T cells showed a significantly greater peak on day 10 but a similar contraction by day 30. Percentages of annexin+ cdd donor CD8+ T cells were significantly reduced compared to the other groups. Persistence of host antigen presenting cells did not account for the unrestrained expansion of cdd donor T cells because host dendritic cells were not detected in either the spleen, BM or gut of recipients of cdd CD8+ T cells on day 6 after BMT. In addition, alloantigen expression on epithelial target cells did not enhance GVHD because B6 donor cdd T cells induced equivalently lethal GVHD in [bm1 → B6] and [bm1 → bm1] chimeras (MST of 30 days and 27 days, respectively). We conclude that both perforin and Fas ligand pathways are required for alloreactive CD8+ T cell populations to contract after their initial expansion during a GVH reaction and that the absence of both these pathways results in donor CD8+ T cell unrestricted expansion and more severe GVHD.
Table 1
GVHD score (gut) IFN-g (pg/ml) TNF-a (pg/ml) CD8+T cell (x10e6) Annexin+CD8+(%) cdd vs. wt, *P&lt;0.05 Wt 4.0±0.4 110±12 6.5±2.7 0.9±0.2 81±3.3 Cdd 5.7±0.3* 263±71* 64.6±3.2* 80.1±4.0* 62±3.6* Pfp−/− ND ND ND 2.6±0.7 73±5.1 Gld ND ND ND 1.7±0.4 80±0.6
Cyclophosphamide Plus Allogeneic CD4+ T Cell Infusion Induces Anti-Lymphoma Immunity Despite Lack of Graft-Versus-Host Disease (GVHD) or Sustained Engraftment.
Authors: Jalla Sanju , Wang Jie , Luznik Leo , Fuchs Ephraim J. 
Published: April 2020

Abstract
Recent evidence suggests that tumor-bearing animals contain CD8+ T cells that can respond productively to a tumor vaccine, but that these T cells do not respond because of insufficient help from tumor-specific CD4+ T cells, which have either been inactivated or turned into anti-tumor suppressor T cells. We therefore devised a strategy to augment anti-tumor immunity by administering cyclophosphamide (Cy), to eliminate suppressor CD4+ T cells, followed by combining autologous tumor cell vaccination and infusion of partially MHC-mismatched, or haploidentical, CD4+ T cells as a source of T cell help for endogenous CD8+ T cells. Interestingly, the combination of Cy followed by haploidentical T cell infusion, with or without vaccine, induced potent systemic anti-tumor immunity resulting in cure of 40-50% of BALB/c mice harboring the A20 B cell leukemia/lymphoma. Depletion of CD8+ T cells from the infusate abrogated GVHD but did not compromise anti-tumor immunity. Allogeneic donor spleen cells that contained CD8+ T cells engrafted durably and caused lethal GVHD. In contrast, the combination of Cy plus CD8+ T cell-depleted spleen cell infusion induced only transient engraftment, peaking on day 7 and declining to undetectable levels by day 14. In the absence of Cy conditioning, allogeneic donor spleen cell infusions did not induce detectable chimerism beyond day 3. In summary, Cy plus allogeneic CD4+ T cell infusion induces potent anti-tumor immunity in a mouse model of B cell leukemia/lymphoma. Potential mechanisms of the therapeutic effect include direct tumor cytotoxicity by CD4+ T cells or allogeneic CD4+ T cell help for endogenous, tumor-specific CD8+ T cells.
Growth Arrest-Specific Gene 6 (Gas6) Deficiency in Mice Alleviates Graft-versus-Host Disease (GVHD) in the Liver.
Authors: Burnier Laurent , Herbert Jean-Marc , Savi Pierre , Collen Désiré , Conway Edward M. , Carmeliet Peter , Izui Shozo , Angelillo-Scherrer Anne 
Published: April 2020

Abstract
GVHD is the single most important complication of allogeneic bone marrow transplantation (BMT), mediated by donor-derived alloreactive T cells. Gas6, a vitamin K-dependent protein, interacts with phospholipid membranes via its γ-carboxyglutamic acid-containing tail and binds to the receptor tyrosine kinases Tyro3, Axl and Mer via its carboxy-terminal globular domain. Gas6 has been implicated in reversible growth arrest, survival, proliferation, cell adhesion, hemostasis and erythropoiesis. Preliminary experiments have shown that Gas6 deficient (Gas6-/-) mice have a leukocyte trafficking defect into inflamed tissues. We hypothesized that donor Gas6-/- T cells might induce less GVHD than wild-type (WT) T cells in MHC-mismatched BMT. We therefore compared the ability of Gas6-/- FVB (H-2q) bone marrow cells to produce GVHD in lethally irradiated MHC haplo-mismatched Gas6-/- and WT B6 mice (H-2b). Gas6-/- and WT B6 received 9 Gy total body irradiation and 107 cells from bone marrow of FVB mice (H-2q) of the same genotype. (Gas6-/- B6 mice received Gas6-/- FVB cells and WT B6 mice were injected with WT FVB cells.) On Day 28, recipients were sacrificed and chimerism was confirmed by expression of H-2q haplotype in recipient bone marrow. Neither group had clinical evidence of GVHD. However, histologic studies revealed lesions in the livers of both genotypes, which were notably less severe in Gas6-/- mice. The size and number of portal space leukocyte infiltrates were reduced in Gas6-/- mice (40 ± 6.6 % of portal spaces infiltrated), compared to WT mice (76.8 ± 6.5 % of portal spaces infiltrated, P &lt; 0.01). Apoptosis of liver cells was also reduced in Gas6-/- mice. We hypothesized that diminished leukocyte trafficking in the Gas6-/- mice may be due to defects in cytokine-induced expression of adhesion molecules. We therefore isolated primary capillary endothelial cells from Gas6-/- and WT mice, and assessed their response to TNF-α. In contrast to WT endothelial cells, Gas6-/- endothelial cells responded without increases in VCAM-1, ECAM-1, E-selectin, IL-1 or IL-6. This resistance to TNF-α was reversed when the Gas6-/- endothelial cells were co-treated with recombinant Gas6, suggesting that the effect of Gas6 on vascular endothelial function is critical for normal leukocyte trafficking in GVHD. Overall, these results provide strong rationale for considering Gas6 and/or its receptors as a potential therapeutic target to minimize GVHD.
Transjugular Liver Biopsy in Hemophilia Is Safe with a Minimum of Two Doses of Factor.
Authors: Ragni Margaret V. , Jaworski Kristen , Boggio Lisa , Brettler Doreen B. , Hoots W. Keith , Kuriakose Philip , Rutherford Cynthia Jean , Zajko Albert B. 
Published: April 2020

Abstract
Over 80% of adults with hemophilia are infected with hepatitis C (HCV) infection through exposure to clotting factor concentrates 20 or more years ago. Although liver biopsy is considered the gold standard to assess the severity of HCV liver disease, the majority of those with hemophilia do not undergo biopsy, even by the less invasive transjugular route. Little is known about the actual or perceived bleeding risks which may affect patient decisions about biopsy. Further, little is known about the minimal hemostatic dose of factor replacement which may affect physician decisions about biopsy. Of 161 hemophilic men enrolled in the multicenter HIV and HCV in Hemophilia (HHH) study which evaluates extent of and risks for HCV disease progression, 112 (69.6%) have decided against biopsy. Among the reasons, given by 75 of the latter, were fear of the procedure, in 23 (30.7%); lack of insurance, in 11 (14.7%); past biopsy, in 11 (14.7%); lack of time, in 10 (13.3%); lack of perceived indication, in 6 (8.0%); presence of an inhibitor, in 5 (6.7%); fear of bleeding, in 5 (6.7%); and other illness or uncertainty, in 4 (5.3%). The 49 (30.4%) agreeing to liver biopsy were more likely than those declining biopsy to be HIV+ (53.1% vs. 34.8%, p &lt; 0.05) and less likely to have an inhibitor (0% vs. 13.4%, p = 0.002), but were as likely to have hemophilia A, 81.6% vs. 83.0%; be over 35 years of age, 49.0% vs. 53.1%; Caucasian, 81.6% vs. 85.7%; and HCV genotype 1, 80.8% vs. 79.5%, all p &gt; 0.05. At the time of biopsy, the median platelet count was 185,000/ul (38,000–366,000/ul), the median PT was 12.0 seconds (9.4–15.4 sec), and the median INR was 1.0 (0.9–1.2). All patients received two doses of factor, and 25 (71.4%) received a third dose; three (8.6%) received a fourth dose for unrelated procedures. The median pre- and post-biopsy F.VIII doses were 49.5 U/kg, 26.6 U/kg at 1–4 hours and 45.0 U/kg at 24–48 hours; and the median F.IX doses were 74.0 U/kg, 44.3 U/kg, and 45.0 U/kg, respectively. No biopsy-related bleeding occurred. Minor adverse events included fever in two, liver function abnormality in one, and duodenal ulcer bleeding in one. The median Knodell, Ishak, and Metavir scores did not differ significantly by HIV status (+ vs.), type hemophilia (A vs. B), age (&gt;35 vs ≤ 35 yr), HCV genotype (1 vs. non-1), or race (Caucasian vs. non-Caucasian). Although preliminary, these findings suggest that fear of the procedure is the most common deterrent to liver biopsy in patients with hemophilia. Yet, when performed by the transjugular route, liver biopsy appears to be safe, with excellent hemostasis achieved with a minimum of two doses of factor, including a 100% dose pre- and a 50% dose post-biopsy. These findings will require confirmation in larger numbers of subjects.
Incidence and Outcome of Discontinuation of Prophylactic Treatment among Young Adults with Severe Hemophilia.
Authors: van Dijk Karin , Fischer Kathelijn , van der Bom Johanna G. , Scheibel Elma , Ingerslev Jørgen , van den Berg H. Marijke 
Published: April 2020

Abstract
Introduction
Most of the current discussion about prophylaxis for severe hemophilia patients is on the dose and when to start. However, as hemophilia is a chronic disease, it is important to evaluate the duration of prophylaxis. The aim of this study was to study and compare adherence to prophylaxis and outcome of severe hemophilia patients.
Methods
All patients with severe hemophilia A and B (factor VIII/IX&lt;0.01 IU/ml), born between 1970 and 1980 and treated in Copenhagen, Århus (Denmark) or the Van Creveldkliniek, Utrecht (The Netherlands) were studied. Data on treatment were collected from the patients’ files from 1972 until 2003. In addition, a questionnaire on adherence to prophylaxis was used. For assessment of outcome the clinical score according to Gilbert and the radiological Pettersson score were used.
Patients were categorized according to adherence to prophylactic treatment: patients who never discontinued prophylaxis (never), patients who temporarily discontinued prophylaxis (temporarily) and patients had switched to on demand regimen (permanently).
Results
83 patients were studied. Median follow up was 19 years (range 6–29). Median age at start of prophylaxis was 5.9 years (interquartile range (IQR) 4.0–8.7). 34% of patients stopped taking prophylaxis temporarily and 35% stopped taking prophylaxis permanently at a median age of 21.5 years (IQR 18.4–24.4). Follow up since the last stop was 3.6 years (IQR 1.4–7.9), the annual number of joint bleeds on on demand treatment was 3.0 (IQR 1.4–8.7).
The median clinical score was 3.0 points (IQR 1.0–6.0) in patients who never or temporarily stopped and 4.0 (IQR 0.0–6.3) in patients who permanently stopped taking prophylaxis. Pettersson scores were available for the Dutch patients and the median Pettersson score was 13 points (IQR 5–23) for patients who never or temporarily stopped and 13 (IQR 1–24) for patients who stopped permanently. The proportion of patients who discontinued prophylaxis and outcome parameters were similar for the Dutch and Danish patients.
Conclusion
Two thirds of young adults with severe hemophilia on prophylaxis discontinue prophylaxis at least once. One third permanently stop taking prophylaxis, while maintaining a low joint bleed frequency. Four years after switching to on demand treatment, outcome in these patients is similar to outcome in patients who continue taking prophylaxis.