bluuebunny/crossref_metadata_2025_split huggingface dataset:
Elevated Fibrinogen Predicts Atherosclerosis.
Authors: Green David , Foiles Nancy , Chan Cheeling , Schreiner Pamela J. , Jacobs David , Liu Kiang 
Published: October 2019

Abstract
Elevated levels of hemostatic factors are observed in patients with atherosclerosis, but whether they promote plaque formation or are a consequence of the disease is uncertain. To examine this issue, we used data from a large biracial cohort of young adults (Coronary Artery Risk Development in Young Adults [CARDIA]) followed up for 13 years, to examine the relationships of hemostatic factors - fibrinogen, factors VII and VIII, and von Willebrand factor (vWF) - with coronary artery calcium (CAC) and carotid intimal-medial thickness (IMT). Complete data were available on 1382 participants, whose mean age was 32 years at enrollment. The age, race, and gender-adjusted prevalence of CAC for increasing quartiles of fibrinogen levels was: 14.0%, 15.0%, 19.6%, and 28.4% (p <0.001 for trend). After further adjustment for BMI, smoking, systolic BP, and total cholesterol, the prevalence of CAC for increasing quartiles of fibrinogen was 15.5%, 16.0%, 19.0%, and 26.4% (p <0.001 for trend). Similar trends were observed for IMT (age, race, and gender-adjusted, p<0.001; multivariable adjusted, p=0.022). When race and gender subgroups were further analyzed, the prevalence of CAC was associated with fibrinogen levels in women and white men after age adjustment, and in women on multivariable analysis. IMT scores adjusted for age were associated with elevated fibrinogen levels in all except black men, and in black women after multivariable adjustment (p=0.003). While the prevalence of CAC was not associated with increasing quartiles of FVII, FVIII, or vWF, IMT scores were associated with elevated FVII on multivariable analysis in white women (p=0.006) and with vWF antigen in white men on age-adjusted (p=0.004) and multivariable analysis (p=0.013). There were no significant associations of hemostatic factors with either the prevalence of CAC or IMT in black men. Participants were categorized as to whether they had 0, 1, or more than 1 hemostatic factors in the highest quartile. After adjustment for age, race, and gender, hemostatic group classification was associated linearly with the prevalence of CAC (p<0.001 for trend) and IMT score (p=0.01 for trend). In conclusion, the main finding from this study is that elevated levels of fibrinogen in persons aged 25 to 37 are associated with the later appearance of subclinical markers of cardiovascular disease. These associations were observed in whites and black women, but not black men. We suggest that atherosclerosis became established during the 13 year observation period, and that increased fibrinogen may have been a contributing factor or a marker for disease development.
Impact of Inherited Thrombophilia on Symptomatic Venous Thrombo-Embolism (VTE) in Children: A Systematic Review & Meta-Analysis of 37 Studies Including 2470 Pediatric Patients.
Authors: Young Guy , Friedrichs Frauke , Chan Anthony , Kenet Gili , Simioni Paolo , van Ommen Helen , Krumpel Anne , Bonduel Marianne , Goldenberg Neil , Male Christophe , Albisetti Manuela , Manco-Johnson Marilyn , Brandao Leo , Nowak-Gottl Ulrike 
Published: October 2019

Abstract
Background: Inherited thrombophilia (IT) has been described as a risk factor for venous thromboembolism (VTE) in children. So far the majority of studies performed in the field were either retrospective or prospective on small numbers of patients. Thus, the results are contradictory or inconclusive mainly due to lack of statistical power. The aim of this study was to better estimate the impact of IT on early VTE onset and recurrence in children as a prerequisite to develop primary and secondary treatment options.
Methods: A systematic search of publications listed in the electronic databases (Pubmed, Medline, EMBASE, Web of Science, The Cochrane Library) up to August 2007 using key words in combination both as MeSH terms and text words, was conducted. Citations were screened by two independent group members and those meeting the inclusion criteria were retained. Articles were included if published after 1990, when pediatric VTE was started to be systematically investigated.
Findings: Twenty case-control and 17 cohort studies from 13 countries met the inclusion criteria. In these studies > 70% of patients had at least one clinical risk factor. The summary odds ratios (OR) and 95% confidence intervals (CI) of included studies under a fixed-effects and random-effects model showed statistically significant associations between the IT traits investigated and VTE onset (table). For the rare event of VTE recurrence, 1227 patients (eight studies) were evaluated: at the present state due to high heterogeneity, a trend towards association with recurrent VTE was found for ≥2 IT traits in the fixed-effects model (0R/CI: 2.8/1.6–4.8).
Interpretation: The present meta-analysis gives evidence that the detection of inherited thrombophilia is clincially meaningful in children with VTE and underlines the importance of a pediatric thrombophilia screening program.
Summary of Data Risk Factors OR/CI:fixed model OR/CI:random model patients/controls 2470/4119 N/A FV G1691A 3.5/2.9–4.2 3.2/2.3–4.4 FII G20210A 2.2/1.5–3.3 2.2/1.5–3.4 Protein C defiiciency 9.8/5.9–16 9.9/6.1–16.1 Protein S deficiency 7.1/3.9–13.2 6.8/3.7–12.7 Antithrombin deficiency 7.9/3.8–16.6 7.3/3.4–15.3 Lipoprotein(a) 4.4/3,2–5.9 4/2.4–6.6 ≥ 2 risk factors 12.6/7.3–21.8 11.6/6.2–20.2
An Ex Vivo Perfusion Model To Study the Treatment of Thrombotic Microangiopathy during Pig-to-Human Xenogenic Kidney Transplantation.
Authors: Ramackers Wolf , Friedrich Lars , Schüttler Wolfgang , Bergmann Sabine , Ganser Arnold , Winkler Michael , Tiede Andreas 
Published: October 2019

Abstract
Early rejection of xenogenic organs is associated with thrombotic microangiopathy and changes of coagulation resembling disseminated intravascular coagulation (DIC). Here, we used an ex vivo perfusion circuit as a model of pig-to-human kidney transplantation to study the nature and treatment of this pathology. Porcine kidneys were obtained following in situ cold perfusion with HTK organ preservation solution and immediately connected to a perfusion circuit containing porcine (“autologous”) or human (“xenogenic”) AB blood supplemented with complement component C1 inhibitor (1 U/ml) and heparin (1 U/ml). Perfusion of porcine kidneys with autologous blood was feasible for >240 min in all experiments. In contrast, perfusion of porcine kidneys with xenogenic human blood was limited by a dramatic increase of flow resistance after 30 to 240 min. Increased concentrations of C3a as a marker for complement activation were associated with early perfusion failure. In addition, a dramatic increase of thrombin-antithrombin complex (TAT) and D Dimer (DD) was observed together with consumption of platelets, fibrinogen and antithrombin (AT). Histological examination demonstrated extensive thrombotic microangiopathy. Supplementation recombinant human activated protein C (rhAPC, 300 ug/l*h, n=3) or recombinant human antithrombin (rhAT, 3 U/ml, n=3) abolished the increase of flow resistance and allowed for a xenogenic kidney survival of >240 min in all experiments. Increase of DD was abolished and the consumption of fibrinogen was abrogated by both treatments as compared to control, whereas the increase of TAT was abolished only by rhAPC. Histological examination revealed no evidence of thrombotic microangiopathy for both treatments as compared to control. In conclusion, the perfusion model introduced here is a suitable tool for studying coagulopathy during early rejection in xenotransplantation. Thrombotic microangiopathy and DIC-like activation of coagulation is associated with an increased flow resistance and failure of perfusion in this model. Pharmacological intervention such as the supplementation of rhAPC or rhAT can be studied using this model and has been shown to prevent coagulopathy and thrombotic microangiopathy.
Correlation between FOXP3 Gene Polymorphisms in Donors, and the Severity of Acute Graft-Versus-Host Disease in Patients after Related Allogeneic Stem Cell Transplantation.
Authors: Perfecto-Avalos Yocanxochitl , Escoboza Jose R. Borbolla , Villela-Martiez Luis M. , Scott Sean P. , Vela-Ojeda Jorge , Gonzalez-Ramella Oscar , Baltazar-Arellano Severiano , Lopez-Hernandez Manuel A. 
Published: October 2019

Abstract
INTRODUCTION: Acute Graft-versus-host disease (aGVHD) is a major complication of allogeneic stem cell transplantation (alloSCT). Risk factors include patient age, sex matching, CMV status and degree of match. Regulatory T cells are critical for immune tolerance processes such as aGVHD, and express the transcription factor FOXP3, a member of the forkhead/winged-helix family, identified as a key regulatory gene required for the development and activity of these cells. It has been suggested that genetic expression of FOXP3 is inversely correlated with the severity of the GVHD. We studied donor’s DNA looking for 5 polymorphisms on the promoter region of the FOXP3 gene, and we tried to correlate them with presence and degree of aGVHD.
PATIENTS AND METHODS: We studied donors of stem cells for allogeneic stem cell transplants. We looked for the presence of the following polymorphisms by PCR: POL01 −5906 T/A rs2869211; POL03 −3279 A/C rs3761548 ; POL04 −2383 C/T rs3761549 ; POL05 −1383 C/T rs2232364 ; POL06 −924 A/G rs2232365.
RESULTS: Our sample consisted of 31 donors, all siblings. In them we found only 2 of the 5 FOXP3 polymorphisms, either as homozygous or heterozygous. These polymorphisms were found in 15/31 donors, with 12 being homozygous (38.7%), and 3 heterozygous (9.7%). These genes polymorphisms were POL03 y POL06. The most observed polymorphism was POL-06 with 9 cases, while POL-03 was found in 6 donors. Only sex difference and CMV status had an elevated hazard ratio for developing GVHD (HR=1.18, CI95%: 0.18 to 7.64; p=0.85) and (HR=3.0, CI95%: 0.07 to 126; p=0.46) respectively. We found no statistically significant difference in the incidence of GVHD between patients who had received cells from donor with or without a FOXP3 polymorphism (p=0.87). When we analyzed the risk of presenting GVHD, results suggest that having one of the 2 positive polymorphisms of the FOXP3 gene could have a protective effect for the patient. For POL03 HR=0.87, CI95%:0.18 to 4.14; p=0.86, and for POL06, HR=1, CI95%: 0.37 to 2.64, p=0.67.
CONCLUSIONS: Even though our sample is still small to make conclusive remarks, we believe that our results point toward some level of “protection” from acute GVHD in patients receiving cells from donors expressing POL03 and POL06. It is also worthwhile mentioning the relatively high frequency of these polymorphisms, as well as the absence of the other 3.
Neutralization of Macrophage Migration Inhibitory Factor (MIF) Reduces GVHD but Preserves GVL.
Authors: Miklos Sandra , Mueller Gunnar , Lindner Petra , Chang Ya-Yi , Holler Ernst , Bucala Richard , Hildebrandt Gerhard C. 
Published: October 2019

Abstract
Acute graft versus host disease (aGVHD) is the major complication after allogeneic (allo) stem cell transplantation (SCT). The pathophysiology of aGVHD involves injury to host tissues by both inflammatory cytokines and donor-derived cellular effectors. Macrophage migration inhibitory factor (MIF) is produced by various cell types and has a broad range of proinflammatory properties including the induction of T cell activation and inflammatory cytokine secretion. In this study, we tested the hypothesis, that MIF contributes to systemic inflammation and mortality after allo-SCT by using a well established murine model (B6→B6D2F1). Lethally (1300cGy) irradiated B6D2F1 (H-2bxd) mice received SCT either from syngeneic (syn) (B6D2F1) or allo (B6; H2-b) donors. One half of animals after allo SCT was treated with polyclonal antibodies against mouse MIF from day 0 until day 14, whereas syn animals and the other half of allo recipients were treated with control IgG. The severity of GVHD was assessed after SCT by survival and a clinical scoring system that incorporates changes in weight loss, fur texture, skin integrity, mobility and posture. As expected, syn-SCT recipients all survived and were indistinguishable from naïve, untransplanted controls, whereas animals receiving allo-SCT plus control IgG developed significant aGVHD and none of the animals survived by day +28. By contrast, in allo recipients treated with anti-MIF antibodies clinical GVHD (day 21: 4.4±0.4 vs. 5.7±0.5) and weight loss (24.6% vs. 45.4%) were less severe, and this reduction in clinical GVHD translated into significantly improved survival (p<0.05). In addition, serum IFNγ (4397±995 vs. 6070±525 pg/ml) and TNFα (113.9±12.7 vs. 159.2±22.5 pg/ml) levels of anti-MIF treated recipients were decreased by day 7. Alloantigen-specific T cell activation in vitro was significantly suppressed in the presence of anti-MIF, as determined by a reduction in T cell proliferation (38517±2411 vs. 65076±3959 cpm), IFNγ (832±117 vs. 4604±179 pg/ml) and TNFα (189.0±7.5 vs. 522.9±6.1 pg/ml) secretion. We next challenged syn and allo recipients with P815 tumor cells (H-2d) at the time of transplantation. No tumor cell elimination was seen after syn SCT. In contrast, FACS analysis revealed, that clearance of P815 cells was comparably high in allo recipients treated with either control or anti-MIF IgG. In summary, our data strongly suggest, that neutralizing MIF may be a novel approach to reduce GVHD but maintain GVL.
Down Regulation of CXC Chemokine Receptor 3 Impedes the Ability of Anti-CXCR3 Antibody To Block Graft-Versus-Host Disease.
Authors: He Shan , Cao Qi , Jin Min , Emerson Stephen G. , Zhang Yi , Zhang Yanyun 
Published: October 2019

Abstract
Graft versus host disease (GVHD) is caused by infused donor T cells that primarily target specific organs, such as skin, liver and intestine. While it has been demonstrated that chemokines are critical to directing in vivo trafficking of host-reactive donor T cells into GVHD target organs, blocking GVHD using antagonist antibody (Ab) against chemokine receptors remains an elusive goal. We found that in vivo administration of anti-CXCR3 Ab for 1 week (named short-term treatment) only partially prevented acute GVHD in B6/SJL mice receiving donor C3H.SW naïve CD8+ T cells, with 45% of them dying from the disease. Flow cytometry analysis showed that CXCR3+ CD8+ T cells occurred in the spleens and livers of these GVHD B6/SJL mice by day 3 and peaked by day 7 after transplantation, and persisted throughout the disease course. When donor CXCR3+ CD8+ T cells were recovered from these GVHD B6/SJL recipients at day 14 after transplantation (named day-14 CXCR3+ CD8+ T cells) and adoptively transferred into secondary B6/SJL mice, they caused severe GVHD in these secondary recipients with 90% mortality. Interestingly, short-term treatment using anti-CXCR3 Ab protected 78% of B6/SJL mice receiving these donor day-14 CXCR3+ CD8+ T cells. Thus, besides CXCR3+ T cells, some other pathogenic T cells have developed during GVH reactions. We found that a substantial population of CXCR3− CD8+ T cells occurred by day 14 (termed day-14 CXCR3− CD8+ T cells) after transplantation in the spleens (71.1%) and livers (81.6%) of B6/SJL recipients with ongoing GVHD. As compared to donor day-14 CXCR3+ CD8+ T cells, donor day-14 CXCR3− CD8+ T cells expressed lower levels of CCR2, CCR3, CCR5, CD127, granzyme B and FasL, and produced less IFN-g and TNF-a. Furthermore, adoptive transfer of donor day-14 CXCR3− CD8+ T cells caused delayed onset of GVHD as compared to those recipients of day-14 CXCR3+ CD8+ T cells, with 55% of them dying from the diseases. Prolonged treatment of B6 mice receiving day-14 CXCR3− CD8+ T cells with anti-CXCR3 Ab up to 3 weeks (named long-term treatment) significantly inhibited the development of GVHD. More than 70% of these recipients survived without clinical signs of GVHD. In vivo experiments showed that 23% of day-14 CXCR3− CD8+ T cells were converted into CXCR3+ by 12 hours after adoptive transfer into B6/SJL recipients, and up to 60% by 24 hours. Interestingly, about 25% of CXCR3+ CD8+ T cells were also converted into CXCR3− by 24 hours after their transfer. We found in ex vivo cultures that both dendritic cells and interleukin-2 accounted for this conversion between CXCR3+ and CXCR3− subsets, which was inhibited by addition of Ab specific to MHC-I or CTLA-4 Ig. Taken together, these results suggest that alloantigen-induced repression of chemokine receptor(s) may present a major barrier to the success of GVHD prevention and treatment when using antagonists against chemokines or chemokine receptors.
Discovery and Validation of STAT-3 and ERK1/2 Phosphorylation as Critical for the Function of Alloactivated T Cells in Acute Graft-Versus-Host-Disease Via a Novel Technique for Drug Discovery.
Authors: Lu Sydney X. , Alpdogan Onder , Campos Roberto , Wang Xiao , Gao Guo-Jian , Chow Melanie , Lin Janine , Suh David , King Christopher , Hubbard Vanessa M. , Bautista Johanne L. , Kochman Adam A. , Zakrzewski Johannes L. , Smith O. Marsinay , Grubin Jeremy , Chow Andrew , Patel Neel , van den Brink Marcel R.M. 
Published: October 2019

Abstract
Graft-versus-host-disease (GVHD) is a serious complication of allogeneic bone marrow transplantation (allo-BMT). The physiology of GVHD is dominated by alloactivated donor T cells, yet current treatments are often nonspecific and offer limited efficacy with relatively high toxicities. We screened for novel drug targets in alloactivated T cells in a murine allo-BMT model using a flow cytometric technique for the in vivo analysis of intracellular signaling. We defined the signaling profile of normal T cells and alloreactive T cells during GVHD, focusing on pathways involved in T cell receptor (TCR), costimulatory, and cytokine signaling. This analysis revealed that although proteins in multiple pathways (MAP kinases, PI3K, Jak/STAT signaling) were all heavily phosphorylated in alloactivated T cells, phosphorylation of STAT-3 and ERK1/2 were particularly prominently increased in donor alloactivated CD4 T cells. We further analyzed the importance of STAT-3 and ERK1/2 signaling in alloactivated T cells via the use of small-molecule inhibitors of STAT-3 (curcurbitacin E/I) and ERK1/2 phosphorylation (SL327). Treatment with these inhibitors attenuated T cell proliferation in response to anti-CD3+CD28 stimulation and in mixed leukocyte reactions in vitro in a dose-dependent fashion (figure 1).
Figure 1 Figure 1.
Pre-incubation of donor splenocytes with cucurbitacin E significanly reduced T cell activation (CD25, CD69) at 24 hours in adoptive transfer experiments in vivo (p<0.05). To rule out any direct toxicity, we analyzed recovered cells for apoptosis by Annexin-V staininga dn detected no significant toxicity (figure 2).
Figure 2 Figure 2.
We conclude that flow cytometric analysis of signaling pathways in single cells represents a novel methodology to assess the in vivo signaling profiles of specific cell populations in order to select drug targets for further study. STAT-3 and ERK1/2 phosphorylation may also represent potential targets to selectively inhibit donor T cell alloactivation and proliferation in GVHD.
Flow Cytometry Analysis of Peripheral Blood CD4+/CD25+/FOXP3+ T Regulator Cells from 11 Patients Treated with Ipilimumab Following Relapse of Malignancy after Allogeneic Hematopoietic Stem Cell Transplantation.
Authors: Zhou Jiehua , Zhong Ruikun , Corringham Sue , Sapp Teresa , Soiffer Robert , Mitrovich Rachel C. , Lowy Izzy , Bashey Asad , Ball Edward D. 
Published: October 2019

Abstract
We conducted a multi-center dose-escalation trial to assess whether CTLA-4 blockade is safe and efficacious as immunotherapy in patients with relapse of malignancy following allogeneic hematopoietic stem cell transplantation (allo-HCT). We hypothesized that since CTLA-4 is a negative regulator of effector T-cell activation, inhibition of CTLA-4 ligation with a blocking human monoclonal antibody (ipilimumab) will augment graft-versus-malignancy. Here we report on the effects of a single dose of ipilimumab on in vivo T cell subsets from 11 patients treated at the highest dose level (3 mg/kg) and compare these findings with those in normal volunteer donor lymphocytes. We analyzed the expression of intracellular CTLA-4 and FOXP3 on CD4+/CD25+ Treg cells, intracellular cytokines and surface markers for T-cell activation on peripheral T cells from 11 patients and 9 normal donors. Peripheral blood mononuclear cells (PBMC) from patients before (day 0) and after the treatment at day 1, 3, 7, 14 and 30 were stained with a panel of antibodies and analyzed by flow cytometry. Lymphocytes from normal donors at time zero and 3 days after culture in IL-2 (200u/ml) were used as controls. The pre-treatment expression of intracellular CTLA-4 was significantly higher in CD4+ T cells from patients than normal controls and was increased further after antibody treatment from 7.1±3.8% at day 0 to 18.2±7.1% at day 30 (p=0.02). Although the expression of FOXP3 in CD4+/CD25+ T cells was higher in patients than in normal donors (6.3±4.8% compared with 2.0±1.6%, p=0.02), there was no significant change in the levels following ipilimumab infusion. The expression of CD4+/CD25high in the patients was 7.7±2.8%, higher than the normal donors (2.3±1.1%). However, 6/11 cases had increased expression while others had decrease or no change, overall there was no statistically significant change. CD4+/CD25low activated T cells were elevated in 10/11 patients before ipilimumab (42.1±8.5%). Their levels were not affected by CTLA-4 blockade. CD8+/CD69+ activated T-cells were significantly increased in 8/11 patients within the 30 days after ipilimumab treatment but typically returned to baseline values on longer follow-up. CD4+/CD69+ and CD4+/HLA-DR+ T-cells were unchanged following ipilimumab treatment. These data show that a single dose of ipilimumab enhances levels of some subsets of activated T cells without a significant effect on cells with a T-regulatory phenotype.
Vβ Spectratype Analysis of Tissue-Infiltrating T Cells Responding to Minor Histocompatibility Antigens Involved in Graft-Versus-Host Disease.
Authors: Zilberberg Jenny , Loise Gichuru N. , Friedman Thea M. 
Published: October 2019

Abstract
Lethal graft-versus-host disease (GVHD) can be induced between MHC-matched murine strains expressing multiple minor histocompatibility antigen (miHA) differences. In the C57BL6 (B6)->BALB.B strain combination, both CD4+ and CD8+ donor T cells can mediate severe lethal GVHD, whereas in the B6->CXB-2 model, in which the CXB-2 strain expresses a subset of the BALB.B miHA, only the CD8+ T cells directly potentiate lethality. We have previously used TCR Vβ CDR3-size spectratype analysis to examine the alloreactive B6 CD4+ and CD8+ T cells, isolated from the lymphohematopoietic compartment after transplantation into both BALB.B and CXB-2 recipients. However, since tissue-specific expression of miHA can potentially elicit differential T cell responses, we have extended our T cell repertoire analysis to examine the responses involved in target tissue damage. Infiltrating host-presensitized B6 CD4+ and CD8+ T cells were isolated post-transplant from the intestines, livers and spleens of lethally irradiated (9 Gy; split-dose) BALB.B and CXB-2 recipients. The results indicated some overlapping Vβ CDR3-size skewing in both the CD4+ and CD8+ T cell repertoires between the BALB.B and CXB-2 recipients within the tissues of each recipient strain. Most notably, spectratype analysis demonstrated tissue specific responses unique to each of the BALB.B and CXB-2 infiltrates. In situ observations of the tissue infiltrating alloreactive T cells were performed by fluorescent microscopy of transplanted B6 T cells constitutively expressing eGFP into BALB.B and CXB-2 recipients, in conjunction with immunohistochemical staining of skewed Vβ families. TUNEL staining was also performed to confirm apoptosis of tissue epithelium. These analyses confirmed the increased infiltration of skewed CD4+ and CD8+ Vβ families within the target tissues.