scholarly journals IL-18 mediates sickle cell cardiomyopathy and ventricular arrhythmias

Blood ◽  
2020 ◽  
Author(s):  
Akash Gupta ◽  
Yu-Dong Fei ◽  
Tae Yun Kim ◽  
An Xie ◽  
Ken Batai ◽  
...  
Keyword(s):  
Gene Expression ◽  
Sickle Cell ◽  
Cardiac Fibrosis ◽  
Mononuclear Cells ◽  
Humanized Mouse Model ◽  
Peripheral Blood Mononuclear ◽  
Patients At Risk ◽  
The Right ◽  
Il18 Gene

Previous reports indicate IL18 is a novel candidate gene for diastolic dysfunction in sickle cell disease (SCD)-related cardiomyopathy. We hypothesize that IL-18 mediates the development of cardiomyopathy and ventricular tachycardia (VT) in SCD. Compared to control (CTR) mice, a "humanized" mouse model of SCD exhibited increased cardiac fibrosis, prolonged action potential duration (APD), higher VT inducibility in vivo, higher cardiac NFκB phosphorylation and circulating IL-18 levels, as well as reduced voltage-gated potassium channel expression, translating to reduced outward potassium current (Ito) in isolated cardiomyocytes. IL-18 administration to isolated mice hearts resulted in VTs, originating from the right ventricle, and further reduced Ito in SCD mice cardiomyocytes. Sustained IL-18 inhibition via IL-18 binding protein resulted in decreased cardiac fibrosis and NFκB phosphorylation, improved diastolic function, normalized electrical remodeling and attenuated IL-18-mediated VT in SCD mice. Patients with SCD and either myocardial fibrosis or increased QTc displayed greater IL18 gene expression in peripheral blood mononuclear cells (PBMC), with QTc strongly correlated with plasma IL-18 levels. PBMC-derived IL18 gene expression was increased in non-surviving over surviving subjects. IL-18 is a mediator of sickle cell cardiomyopathy and VT in mice and a novel therapeutic target in patients at risk for sudden death.

scholarly journals Establishment of a Novel Humanized Mouse Model To Investigate In Vivo Activation and Depletion of Patient-Derived HIV Latent Reservoirs

2019 ◽  
Vol 93 (6) ◽  
Author(s):  
Nina C. Flerin ◽  
Ariola Bardhi ◽  
Jian Hua Zheng ◽  
Maria Korom ◽  
Joy Folkvord ◽  
...  
Keyword(s):  
Mouse Model ◽  
Hiv Infection ◽  
Mononuclear Cells ◽  
Systemic Infection ◽  
Humanized Mouse Model ◽  
Peripheral Blood Mononuclear ◽  
Immunodeficient Mice ◽  
Latently Infected ◽  
Infected Cells

ABSTRACT Curing HIV infection has been thwarted by the persistent reservoir of latently infected CD4+ T cells, which reinitiate systemic infection after antiretroviral therapy (ART) interruption. To evaluate reservoir depletion strategies, we developed a novel preclinical in vivo model consisting of immunodeficient mice intrasplenically injected with peripheral blood mononuclear cells (PBMC) from long-term ART-suppressed HIV-infected donors. In the absence of ART, these mice developed rebound viremia which, 2 weeks after PBMC injection, was 1,000-fold higher (mean = 9,229,281 HIV copies/ml) in mice injected intrasplenically than in mice injected intraperitoneally (mean = 6,838 HIV copies/ml) or intravenously (mean = 591 HIV copies/ml). One week after intrasplenic PBMC injection, in situ hybridization of the spleen demonstrated extensive disseminated HIV infection, likely initiated from in vivo-reactivated primary latently infected cells. The time to viremia was delayed significantly by treatment with a broadly neutralizing antibody, 10-1074, compared to treatment with 10-1074-FcRnull, suggesting that 10-1074 mobilized Fc-mediated effector mechanisms to deplete the replication-competent reservoir. This was supported by phylogenetic analysis of Env sequences from viral-outgrowth cultures and untreated, 10-1074-treated, or 10-1074-FcRnull-treated mice. The predominant sequence cluster detected in viral-outgrowth cultures and untreated mouse plasma was significantly reduced in the plasma of 10-1074-treated mice, whereas two new clusters emerged that were not detected in viral-outgrowth cultures or plasma from untreated mice. These new clusters lacked mutations associated with 10-1074 resistance. Taken together, these data indicated that 10-1074 treatment depletes the reservoir of latently infected cells harboring replication competent HIV. Furthermore, this mouse model represents a new in vivo approach for the preclinical evaluation of new HIV cure strategies. IMPORTANCE Sustained remission of HIV infection is prevented by a persistent reservoir of latently infected cells capable of reinitiating systemic infection and viremia. To evaluate strategies to reactivate and deplete this reservoir, we developed and characterized a new humanized mouse model consisting of highly immunodeficient mice intrasplenically injected with peripheral blood mononuclear cells from long-term ART-suppressed HIV-infected donors. Reactivation and dissemination of HIV infection was visualized in the mouse spleens in parallel with the onset of viremia. The applicability of this model for evaluating reservoir depletion treatments was demonstrated by establishing, through delayed time to viremia and phylogenetic analysis of plasma virus, that treatment of these humanized mice with a broadly neutralizing antibody, 10-1074, depleted the patient-derived population of latently infected cells. This mouse model represents a new in vivo approach for the preclinical evaluation of new HIV cure strategies.

Alterations In HLA-DR Expression In Peripheral Blood Mononuclear Cells Are Associated with An Elevated Tricuspid Regurgitant Jet Velocity and Pulmonary Hypertension of Sickle Cell Disease

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 2640-2640
Author(s):  
Elizabeth Klings ◽  
Daniel A Dworkis ◽  
Surinder Safaya ◽  
Guihua Li ◽  
Lynda Reid ◽  
...  
Keyword(s):  
Gene Expression ◽  
Pulmonary Hypertension ◽  
Sickle Cell ◽  
Peripheral Blood Mononuclear Cells ◽  
Mortality Risk ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Leg Ulcers ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells

Abstract Abstract 2640 Sickle cell anemia (SCA) is characterized by a chronic inflammatory vasculopathy. An elevated tricuspid regurgitant jet velocity (TRV) by echocardiography in SCA patients is not only reflective of possible pulmonary hypertension but is also associated with increased mortality risk, relative systemic hypertension, and renal disease, all of which suggest this is a marker of a more diffuse vasculopathy. We hypothesized that differential gene expression in peripheral blood mononuclear cells (PBMCs) occurs in SCA patients with an elevated TRV and might reflect the role of inflammation in sickle vasculopathy. Peripheral blood samples were obtained from 15 steady-state SCA patients (7 of whom had a TRV > 2.5 m/sec). Three cases with increased TRV had right heart catheterization that confirmed the presence of pulmonary hypertension. PBMCs were isolated via a Ficoll separation. RNA was isolated and amplified using the Ambion Illumina Total RNA Prep Kit, and then hybridized to Illumina Human HT-12 V3 Expression Bead Chips and scanned. Expression data was background corrected, log-transformed, and strictly filtered to consider only probes showing expression in each sample and significant expression in at least half of either group of samples. Differential expression was tested for significance using a t-test performed on both native and rescaled datasets, and only probes reaching a significance of p < 0.05 in both datasets were considered further. Of the 50 probes selected as significant in this way, the two of the most interesting were HLA-DRB1 and MYOM2, observed almost exclusively in SCA patients with a normal echocardiogram (p=0.007 and 0.008 respectively). Differential PBMC gene expression across the HLA-DRB1 locus has been associated in other populations with impaired immunity and increased susceptibility to sepsis. Our previous genetic studies have identified a potential link between the HLA loci and leg ulcers, another hemolytic complication associated with an elevated TRV in 219 SCA cases when compared with 1180 controls. In those studies, HLA-B was found to be in linkage disequilibrium with 3 genes found to be significantly associated with leg ulcers. This suggests the possibility that these heritable loci may play a role in disease modulation in SCD. PBMCs in SCA patients might have a pro-inflammatory phenotype and this may play a role in the vasculopathy and increased mortality risk observed in these patients. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Gene expression of FOXP3, indoleamine 2,3-dioxygenase (IDO), IL10 and CSF1 in the uterus of cows that received an intrauterine infusion of maternal and paternal antigens

2020 ◽  
Vol 36 (6) ◽  
Author(s):  
Talita Vieta Junqueira ◽  
Nadia Simarro Fagundes ◽  
Marcelo José Barbosa Silva ◽  
João Paulo Elsen Saut ◽  
Marcelo Emílio Beletti
Keyword(s):  
Gene Expression ◽  
Reproductive Performance ◽  
Mononuclear Cells ◽  
Peripheral Blood Mononuclear ◽  
Maternal Acceptance ◽  
Immunoregulatory Cells ◽  
Favorable Conditions ◽  
Maternal Tolerance ◽  
Uterine Body

Sensitization with conceptus antigens has been shown to be useful for improving reproductive performance facilitating maternal acceptance of an allogeneic embryo through the induction of cytokines and immunoregulatory cells in the uterine microenvironment. As FOXP3, IDO, IL10 and CSF1 in the uterus are important on the recognition and development of embryos during early pregnancy, this study aimed to determine whether simultaneous or isolated administration of paternal (semen) and maternal (PBMCs) antigens in the uterus of cow, on the day of estrus, influence the gene expression of these cytokines. Forty crossbred cows were divided into four treatments: T0: Control; T1: Semen; T2: PBMCs (peripheral blood mononuclear cells) from another cow and T3: PBMCs+Semen. Antigens were administered into the uterine body on the estrus day (D0). Uterine biopsies designed for molecular analysis of gene expression were collected in vivo seven (D7) and fourteen (D14) days after immunostimulation. Transcripts from FOXP3, IDO, IL-10 and CSF-1 were detected in all RNA samples extracted from uterine biopsies. The semiquantitative analysis showed that none of the treatments caused significant increase in the expression of these genes. Furthermore, on D14 all treatments led to a decline in the number of CSF-1 transcripts; moreover, treatment with PBMCs+Semen also led to a drop in the abundance of IL-10 transcripts. Such results suggest that isolated or simultaneous administration of both antigens would not increase maternal tolerance to embryo alloantigens, nor would it create favorable conditions to its growth and pre-implantation development, at least regarding the effects mediated by these genes on D7 and D14 of the estrous cycle.

A mini-review: microRNA in arthritis

2011 ◽  
Vol 43 (10) ◽  
pp. 566-570 ◽  
Author(s):  
Tomoyuki Nakasa ◽  
Yoshihiko Nagata ◽  
Keiichiro Yamasaki ◽  
Mitsuo Ochi
Keyword(s):  
Gene Expression ◽  
Noncoding Rna ◽  
Mononuclear Cells ◽  
Expression Patterns ◽  
Joint Destruction ◽  
Synovial Cell ◽  
Specific Expression ◽  
Peripheral Blood Mononuclear ◽  
Altered Expression

MicroRNA (miRNA) is a class of noncoding RNA that exhibits tissue- or developmental stage-specific expression patterns and negatively regulates gene expression. MiRNAs play an important role in human diseases, including osteoarthritis (OA) and rheumatoid arthritis (RA). OA is characterized by the progressive destruction of articular cartilage, and several miRNAs exhibit altered expression, playing a role in regulating gene expression in OA pathogenesis, especially in catabolic factors such as matrix metalloproteinases (MMP) and aggrecanases. RA is an autoimmune disease that is characterized by irreversible joint destruction due to chronic synovial inflammation. MiRNAs play an important role in inflammatory response, synovial cell proliferation, and production of MMPs in RA synovial tissues. The expression level of several miRNAs in peripheral blood mononuclear cells correlates with RA disease activity. Recently, therapeutic trials aimed at targeting miRNA in vivo have been conducted. Targeting miRNA will enable a new advanced strategy toward arthritis treatment.

Preliminary Report of In vitro and In vivo Effectiveness of Dornase Alfa on SARS-CoV-2 Infection (Preprint)

2020 ◽  
Author(s):  
Hacer Kuzu Okur ◽  
Koray Yalcin ◽  
Cihan Tastan ◽  
Sevda Demir ◽  
Bulut Yurtsever ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Respiratory Tract ◽  
Mononuclear Cells ◽  
Multiple Organ ◽  
Peripheral Blood Mononuclear ◽  
Dornase Alfa ◽  
Tract Infections ◽  
Adult Peripheral Blood

UNSTRUCTURED Dornase alfa, the recombinant form of the human DNase I enzyme, breaks down neutrophil extracellular traps (NET) that include a vast amount of DNA fragments, histones, microbicidal proteins and oxidant enzymes released from necrotic neutrophils in the highly viscous mucus of cystic fibrosis patients. Dornase alfa has been used for decades in patients with cystic fibrosis to reduce the viscoelasticity of respiratory tract secretions, to decrease the severity of respiratory tract infections, and to improve lung function. Previous studies have linked abnormal NET formations to lung diseases, especially to acute respiratory distress syndrome (ARDS). Coronavirus disease 2019 (COVID-19) pandemic affected more than two million people over the world, resulting in unprecedented health, social and economic crises. The COVID-19, viral pneumonia that progresses to ARDS and even multiple organ failure, is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). High blood neutrophil levels are an early indicator of SARS-CoV-2 infection and predict severe respiratory diseases. A similar mucus structure is detected in COVID-19 patients due to the accumulation of excessive NET in the lungs. Here, we show our preliminary results with dornase alfa that may have an in-vitro anti-viral effect against SARS-CoV-2 infection in a bovine kidney cell line, MDBK without drug toxicity on healthy adult peripheral blood mononuclear cells. In this preliminary study, we also showed that dornase alfa can promote clearance of NET formation in both an in-vitro and three COVID-19 cases who showed clinical improvement in radiological analysis (2-of-3 cases), oxygen saturation (SpO2), respiratory rate, disappearing of dyspnea and coughing.

scholarly journals Expression Profiling of Transcriptome and Its Associated Disease Risk in Yang Deficiency Constitution of Healthy Subjects

2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
Ruoxi Yu ◽  
Yin Yang ◽  
Yuanyuan Han ◽  
Pengwei Hou ◽  
Yingshuai Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Healthy Subjects ◽  
Mononuclear Cells ◽  
Clinical Medicine ◽  
Disease Risk ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Molecular Evidence ◽  
Chinese Han ◽  
Peripheral Blood Mononuclear

Objectives. Differences among healthy subjects and associated disease risks are of substantial interest in clinical medicine. According to the theory of “constitution-disease correlation” in traditional Chinese medicine, we try to find out if there is any connection between intolerance of cold in Yang deficiency constitution and molecular evidence and if there is any gene expression basis in specific disorders. Methods. Peripheral blood mononuclear cells were collected from Chinese Han individuals with Yang deficiency constitution (n=20) and balanced constitution (n=8) (aged 18–28) and global gene expression profiles were determined between them using the Affymetrix HG-U133 Plus 2.0 array. Results. The results showed that when the fold change was ≥1.2 and q ≤ 0.05, 909 genes were upregulated in the Yang deficiency constitution, while 1189 genes were downregulated. According to our research differential genes found in Yang deficiency constitution were usually related to lower immunity, metabolic disorders, and cancer tendency. Conclusion. Gene expression disturbance exists in Yang deficiency constitution, which corresponds to the concept of constitution and gene classification. It also suggests people with Yang deficiency constitution are susceptible to autoimmune diseases, enteritis, arthritis, metabolism disorders, and cancer, which provides molecular evidence for the theory of “constitution-disease correlation.”

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