scholarly journals TNF-αDysregulation in Asthma: Relationship to Ongoing Corticosteroid Therapy

2000 ◽  
Vol 7 (3) ◽  
pp. 229-237 ◽  
Author(s):  
Susan Waserman ◽  
Jerry Dolovich ◽  
Mary Conway ◽  
Jean S Marshall
Keyword(s):  
Corticosteroid Therapy ◽  
Mononuclear Cells ◽  
Inhaled Corticosteroids ◽  
Challenge Test ◽  
High Dose ◽  
Peripheral Blood Mononuclear ◽  
Necrosis Factor Alpha ◽  
Significant Difference ◽  
Age And Sex ◽  
Medium Dose

BACKGROUND:Tumour necrosis factor-alpha (TNF-α) is a major proinflammatory cytokine that is thought to be important in the pathogenesis of asthma. However, alterations in systemic regulation of this cytokine in asthma have not been examined in the context of corticosteroid therapy.OBJECTIVES:To examine the ability of peripheral blood mononuclear cells (PBMC) from three different groups of patients with asthma requiring varying amounts of inhaled corticosteroids (ICS) for clinical control, and to examine cells from age- and sex-matched nonasthmatic patients to produce TNF-α.DESIGN:All patients with asthma had a positive methacholine challenge test. 'High dose' ICS patients with asthma required ICS greater than or equal to 800 µg/day. 'Medium dose' patients with asthma were on less than or equal to 500 µg/day of ICS, whereas 'no ICS' patients with asthma had received no ICS for at least three months. Each patient with asthma was examined in parallel with an age- and sex-matched, nonasthmatic, nonatopic control subject. Cells were cultured (with or without the addition of potential stimulators phytohemagglutinin, lipopolysaccharide, formyl-methionine-leucine-phenylalanine or antihuman CD3), and TNF-αproduction was assessed by ELISA.MAIN RESULTS:PBMC from both high dose ICS (n=8) and no ICS (n=11) patients with asthma produced more than twice the amount of TNF-αthan cells from matched nonasthmatic control patients (P<0.01) when cultured alone or in the presence of each stimulus (P<0.05). In contrast, there was no significant difference in TNF-αproduction between medium dose ICS patients with asthma and control patients. A group of asymptomatic atopic patients (n=6) did not have an increased level of TNF-αproduction.CONCLUSIONS:Increases in TNF-αproduction within the PBMC compartment can be observed in both patients with asthma receiving high dose ICS and in a group of patients with mild asthma receiving no ICS therapy, but not in patients with asthma receiving a medium dose of ICS or atopic patients.

Predicting bortezomib-related severe neurologic adverse events by measuring proteasome activity in PBMCs.

2012 ◽  
Vol 30 (15_suppl) ◽  
pp. 8094-8094
Author(s):  
Yukiko Cho ◽  
Mitsuo Hori ◽  
Yasushi Okoshi ◽  
Takuya Komeno ◽  
Chikashi Yoshida ◽  
...  
Keyword(s):  
Proteasome Inhibitor ◽  
Mononuclear Cells ◽  
Proteasome Activity ◽  
Specific Substrate ◽  
Pretreatment Level ◽  
Peripheral Blood Mononuclear ◽  
Treatment Groups ◽  
Significant Difference ◽  
Blood Mononuclear Cells ◽  
Age And Sex

8094 Background: Although a proteasome inhibitor bortezomib (BOR) is one of the backbone drugs for the treatment of multiple myeloma, severe neurologic adverse event (sNE) is a major obstacle for continuing the treatment. As there is no clinically available biomarker for predicting the occurrence of sNAE, we measured the proteasome activity (PrsAtv) in peripheral blood mononuclear cells (PBMCs). Methods: Patients (Pts) were treated by either standard Q3wks or weekly (BOR 1.3mg/m2: day 1, 8, 15, 22; Q5wks) schedule depending on physician’s decision. PBMCs were collected from 21 Pts and 99 healthy volunteers. Pts’ samples were collected before treatment, 1 hour after the 1st injection of BOR, and at the end of the 1st course. PrsAtv in PBMCs was measured by using a specific substrate SUC-LLVY-AMC, which becomes fluorescent upon cleavage by proteasome. Results: Levels of PrsAtv among volunteers were highly variable with no certain correlation with age and sex. Pretreatment levels of PrsAtv among Pts were also variable, and didn't correlate with the occurrence of sNAE. After 1 hour of BOR injection, PrsAtv decreased to 33.2±20.5% (mean ± SD) of the pretreatment level, and it generally recovered (112.5±77.6%) at the end of the 1st course. However, in 12 out of 21 Pts, it didn’t recover to ≥ 100%. Among these Pts, 5 manifested with sNAE (≥G3) during the 2nd or 3rd course of the chemotherapy. In a sharp contrast, no patients whose PrsAtv recovered to ≥100% manifested with sNAE. Pts who manifested with sNAE (5 Pts) showed the lower levels of PrsAtv at the end of the 1st course than those without sNAE (16 Pts) (67.4±11.8 vs 126.6±83.8%, p=0.075). It should be also strengthened that all the Pts who manifested with sNAE were treated by the standard Q3wks schedule, although levels of PrsAtv at the end of the 1st course were not significantly different between Q3 and Q5wks groups (109.0±68.3 vs 121.1±96.4 %, p=0.38). Contrary, there was no significant difference of the bortezomib response between these two treatment groups. Conclusions: Pts whose PrsAtv does not recover to ≥100% at the end of the 1st course are at high risk of manifesting with sNAE, and these Pts should not be treated by the standard Q3wks schedule in the subsequent courses.

scholarly journals Reactivation of Latent Human Cytomegalovirus in CD14+ Monocytes Is Differentiation Dependent

2001 ◽  
Vol 75 (16) ◽  
pp. 7543-7554 ◽  
Author(s):  
Cecilia Söderberg-Nauclér ◽  
Daniel N. Streblow ◽  
Kenneth N. Fish ◽  
Justine Allan-Yorke ◽  
Patricia P. Smith ◽  
...  
Keyword(s):  
T Cells ◽  
Human Cytomegalovirus ◽  
Mononuclear Cells ◽  
Interleukin 1 ◽  
Hla Class I ◽  
Peripheral Blood Mononuclear ◽  
Necrosis Factor Alpha ◽  
Significant Difference ◽  
Hcmv Replication ◽  
Ifn Γ

ABSTRACT We have previously demonstrated reactivation of latent human cytomegalovirus (HCMV) in myeloid lineage cells obtained from healthy donors. Virus was obtained from allogenically stimulated monocyte-derived macrophages (Allo-MDM), but not from macrophages differentiated by mitogenic stimulation (ConA-MDM). In the present study, the cellular and cytokine components essential for HCMV replication and reactivation were examined in Allo-MDM. The importance of both CD4+ and CD8+ T cells in the generation of HCMV-permissive Allo-MDM was demonstrated by negative selection or blocking experiments using antibodies directed against both HLA class I and HLA class II molecules. Interestingly, contact of monocytes with CD4 or CD8 T cells was not essential for reactivation of HCMV, since virus was observed in macrophages derived from CD14+monocytes stimulated by supernatants produced by allogeneic stimulation of peripheral blood mononuclear cells. Examination of the cytokines produced in Allo-MDM and ConA-MDM cultures indicated a significant difference in the kinetics of production and quantity of these factors. Further examination of the cytokines essential for the generation of HCMV-permissive Allo-MDM identified gamma interferon (IFN-γ) but not interleukin-1 or -2, tumor necrosis factor alpha, or granulocyte-macrophage colony-stimulating factor as critical components in the generation of these macrophages. In addition, although IFN-γ was crucial for reactivation of latent HCMV, addition of IFN-γ to unstimulated macrophage cultures was insufficient to reactivate virus. Thus, this study characterizes two distinct monocyte-derived cell types which can be distinguished by their ability to reactivate and support HCMV replication and identifies the critical importance of IFN-γ in the reactivation of HCMV.

Azithromycin Concentrations During Long-Term Regimen in Patients With MALT Lymphoma

2021 ◽  
Author(s):  
Raphael Scheibenpflug ◽  
Markus Obermüller ◽  
Gottfried Reznicek ◽  
Ortrun Neuper ◽  
Wolfgang W. Lamm ◽  
...  
Keyword(s):  
Body Mass Index ◽  
Body Mass ◽  
Malt Lymphoma ◽  
Plasma Levels ◽  
High Performance ◽  
Mononuclear Cells ◽  
High Dose ◽  
Peripheral Blood Mononuclear ◽  
Significant Difference ◽  
Intracellular Concentrations

Abstract IntroductionIn view of the antineoplastic effects of the macrolide clarithromycin in mucosa associated lymphatic tissue (MALT)-lymphoma, we performed a pilot study assessing levels of azithromycin in plasma, peripheral blood mononuclear cells (PBMC) and polymorphonuclear leukocytes (PMN) of MALT-lymphoma patients to determine the pharmacokinetics and potential influences of respective concentrations on the therapeutic outcome.Material and methodsIn total 16 patients with MALT-lymphoma received 1.5g of oral azithromycin once-weekly over 6 months. Blood was sampled directly prior to the following dose every 4 weeks during treatment. Drug levels were analysed by high performance liquid chromatography in plasma and intracellularly in PBMC and PMN. They were correlated with patients’ age, weight and body-mass-index and compared between patients responsive or unresponsive to treatment.ResultsMean azithromycin plasma levels of all patients were 58.97±30.48 ng/ml, remaining stable throughout the treatment period. Correlation analysis of plasma azithromycin showed no significance. Intracellular PBMC concentrations were 6648±8479 ng/ml, without any significant difference between responders and non-responders. Mean PMN levels were 39274±25659 ng/ml and significantly higher in patients unresponsive to treatment (t=2.858, p=0.017).ConclusionOur drug regime led to continuously high plasma and exceedingly high intracellular concentrations of azithromycin in PBMC and PMN. Age, weight or body-mass-index had no significant influence on plasma levels and thence should not be considered in dosage finding. High AZM levels in PBMC did not lead to a better treatment response, whereas enrichment in PMN suggested a poorer outcome. The threshold for immunomodulatory effects on lymphoma cells might not have been reached. Additionally, the finding of stable plasma and intracellular concentrations over months with high-dose azithromycin administered in intervals might also be important for the further design of azithromycin-based trials against MALT-lymphoma.

scholarly journals Lipopolysaccharides prime whole human blood and isolated neutrophils for the increased synthesis of 5-lipoxygenase products by enhancing arachidonic acid availability: involvement of the CD14 antigen.

1993 ◽  
Vol 178 (4) ◽  
pp. 1347-1355 ◽  
Author(s):  
M E Surette ◽  
R Palmantier ◽  
J Gosselin ◽  
P Borgeat
Keyword(s):  
Arachidonic Acid ◽  
Mononuclear Cells ◽  
Leukotriene B4 ◽  
Eicosatetraenoic Acid ◽  
Tnf Alpha ◽  
Peripheral Blood Mononuclear ◽  
Necrosis Factor Alpha ◽  
Factor Alpha ◽  
Priming Activity

Stimulation of heparinized blood with 1 microM formyl-methionyl-leucyl-phenylalanine (FMLP) resulted in the formation of &lt; 30 pmol/ml plasma of 5-lipoxygenase (5-LO) products. The preincubation of blood with 1 microgram/ml of lipopolysaccharide (LPS) (Escherichia coli 0111-B4) for 30 min before stimulation with FMLP resulted in the accumulation of 250-300 pmol of 5-LO products per ml plasma. The major products detected were leukotriene B4 and (5S)-hydroxy-6,8,11,14-eicosatetraenoic acid which were produced in equivalent amounts. The priming activity was detectable with as little as 1-10 ng LPS per ml blood and was optimal using 1-10 micrograms LPS/ml blood. The priming for 5-LO product synthesis was optimal after 20-30 min of preincubation with LPS and declined at preincubation times &gt; 30 min. The priming effect of LPS was also observed using the complement fragment C5a or interleukin 8 as agonists. Polymorphonuclear leukocytes (PMN) and peripheral blood mononuclear cells accounted for 80 and 20% of the synthesis of 5-LO products, respectively. The ability of LPS to prime isolated PMN was dependent on the presence of plasma and was inhibited by the anti-CD14 antibody IOM2, indicating a CD14-dependent priming mechanism. The priming of whole blood with tumor necrosis factor alpha (TNF-alpha) and LPS was additive and the presence of mononuclear cells did not enhance the ability of LPS to prime PMN, indicating that the priming activity of LPS is independent of LPS-induced TNF-alpha synthesis. The mechanism by which LPS enhance 5-LO product synthesis in PMN was investigated. Treatment of PMN with LPS strongly enhanced the release of arachidonic acid after stimulation with FMLP. The release of arachidonic acid was optimal 2-3 min after stimulation with FMLP, attaining levels 5-15-fold greater than those observed in unprimed cells stimulated with FMLP. These results demonstrate that LPS dramatically increases the ability of blood to generate 5-LO products, and support the putative role of leukotrienes in pathological states involving LPS.

scholarly journals Cholesterol Transporters ABCA1 and ABCG1 Gene Expression in Peripheral Blood Mononuclear Cells in Patients with Metabolic Syndrome

Cholesterol ◽  
2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Zahra Tavoosi ◽  
Hemen Moradi-Sardareh ◽  
Massoud Saidijam ◽  
Reza Yadegarazari ◽  
Shiva Borzuei ◽  
...  
Keyword(s):  
Gene Expression ◽  
Metabolic Syndrome ◽  
Mononuclear Cells ◽  
Fasting Blood Glucose ◽  
Control Group ◽  
Regulation Mechanism ◽  
Healthy Individuals ◽  
Peripheral Blood Mononuclear ◽  
Significant Difference ◽  
Abca1 Expression

ABCA1 and ABCG1 genes encode the cholesterol transporter proteins that play a key role in cholesterol and phospholipids homeostasis. This study was aimed at evaluating and comparing ABCA1 and ABCG1 genes expression in metabolic syndrome patients and healthy individuals. This case-control study was performed on 36 patients with metabolic syndrome and the same number of healthy individuals in Hamadan (west of Iran) during 2013-2014. Total RNA was extracted from mononuclear cells and purified using RNeasy Mini Kit column. The expression of ABCA1 and ABCG1 genes was performed by qRT-PCR. Lipid profile and fasting blood glucose were measured using colorimetric procedures. ABCG1 expression in metabolic syndrome patients was significantly lower (about 75%) compared to that of control group, while for ABCA1 expression, there was no significant difference between the two studied groups. Comparison of other parameters such as HDL-C, FBS, BMI, waist circumference, and systolic and diastolic blood pressure between metabolic syndrome patients and healthy individuals showed significant differences (P<0.05). Decrease in ABCG1 expression in metabolic syndrome patients compared to healthy individuals suggests that hyperglycemia, related metabolites, and hyperlipidemia over the transporter capacity resulted in decreased expression of ABCG1. Absence of a significant change in ABCA1 gene expression between two groups can indicate a different regulation mechanism for ABCA1 expression.

scholarly journals A 70-Kilodalton Recombinant Heat Shock Protein ofCandida albicans Is Highly Immunogenic and Enhances Systemic Murine Candidiasis

1998 ◽  
Vol 66 (5) ◽  
pp. 2154-2162 ◽  
Author(s):  
Carla Bromuro ◽  
Roberto La Valle ◽  
Silvia Sandini ◽  
Francesca Urbani ◽  
Clara M. Ausiello ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Mononuclear Cells ◽  
Cell Mediated Immunity ◽  
Healthy Human ◽  
Peripheral Blood Mononuclear ◽  
Necrosis Factor Alpha ◽  
Ifn Γ

ABSTRACT The 70-kDa recombinant Candida albicans heat shock protein (CaHsp70) and its 21-kDa C-terminal and 28-kDa N-terminal fragments (CaHsp70-Cter and CaHsp70-Nter, respectively) were studied for their immunogenicity, including proinflammatory cytokine induction in vitro and in vivo, and protection in a murine model of hematogenous candidiasis. The whole protein and its two fragments were strong inducers of both antibody (Ab; immunoglobulin G1 [IgG1] and IgG2b were the prevalent isotypes) and cell-mediated immunity (CMI) responses in mice. CaHsp70 preparations were also recognized as CMI targets by peripheral blood mononuclear cells of healthy human subjects. Inoculation of CaHsp70 preparations into immunized mice induced rapid production of interleukin-6 (IL-6) and tumor necrosis factor alpha, peaking at 2 to 5 h and declining within 24 h. CaHsp70 and CaHsp70-Cter also induced gamma interferon (IFN-γ), IL-12, and IL-10 but not IL-4 production by CD4+ lymphocytes cocultured with splenic accessory cells from nonimmunized mice. In particular, the production of IFN-γ was equal if not superior to that induced in the same cells by whole, heat-inactivated fungal cells or the mitogenic lectin concanavalin A. In immunized mice, however, IL-4 but not IL-12 was produced in addition to IFN-γ upon in vitro stimulation of CD4+ cells with CaHsp70 and CaHsp70-Cter. These animals showed a decreased median survival time compared to nonimmunized mice, and their mortality was strictly associated with organ invasion by fungal hyphae. Their enhanced susceptibility was attributable to the immunization state, as it did not occur in congenitally athymic nude mice, which were unable to raise either Ab or CMI responses to CaHsp70 preparations. Together, our data demonstrate the elevated immunogenicity of CaHsp70, with which, however, no protection against but rather some enhancement of Candida infection seemed to occur in the mouse model used.

scholarly journals Intrathymic Plasmablasts Are Affected in Patients With Myasthenia Gravis With Active Disease

2021 ◽  
Vol 8 (6) ◽  
pp. e1087
Author(s):  
Yohei Yamamoto ◽  
Naoko Matsui ◽  
Akiyuki Uzawa ◽  
Yukiko Ozawa ◽  
Tetsuya Kanai ◽  
...  
Keyword(s):  
B Cells ◽  
Myasthenia Gravis ◽  
Disease Activity ◽  
Mononuclear Cells ◽  
Young Patients ◽  
B Lymphopoiesis ◽  
Peripheral Blood Mononuclear ◽  
Follicular Helper ◽  
Significant Difference ◽  
Cxcr5 Expression

Background and ObjectivesTo investigate intrathymic B lymphopoiesis in patients with myasthenia gravis (MG) and explore thymus pathology associated with clinical impact.MethodsThymic lymphocytes from 15 young patients without MG, 22 adult patients without MG, 14 patients with MG without thymoma, and 11 patients with MG with thymoma were subjected to flow cytometry analysis of T follicular helper (Tfh), naive B, memory B, plasmablasts, CD19+B220high thymic B cells, B-cell activating factor receptor, and C-X-C chemokine receptor 5 (CXCR5). Peripheral blood mononuclear cells of 16 healthy subjects and 21 untreated patients with MG were also analyzed. Immunologic values were compared, and correlations between relevant values and clinical parameters were evaluated.ResultsThe frequencies of circulating and intrathymic plasmablasts were significantly higher in patients with MG than controls. On the other hand, the frequency of CD19+B220high thymic B cells was not increased in MG thymus. We observed a significant increase in CXCR5 expression on plasmablasts in MG thymus and an increased frequency of intrathymic plasmablasts that was correlated with preoperative disease activity. The frequency of intrathymic Tfh cells was significantly lower in patients who received immunosuppressive (IS) therapy than those without IS therapy. However, there was no significant difference in the frequency of intrathymic plasmablasts irrespective of IS therapy.DiscussionOur findings confirmed a correlation between increased frequency of intrathymic plasmablasts and disease activity before thymectomy. We postulate that activated intrathymic plasmablasts endow pathogenic capacity in MG.

scholarly journals Measles Virus Spread and Pathogenesis in Genetically Modified Mice

1998 ◽  
Vol 72 (9) ◽  
pp. 7420-7427 ◽  
Author(s):  
Branka Mrkic ◽  
Jovan Pavlovic ◽  
Thomas Rülicke ◽  
Pietro Volpe ◽  
Christian J. Buchholz ◽  
...  
Keyword(s):  
Measles Virus ◽  
Mononuclear Cells ◽  
High Dose ◽  
Peripheral Blood Mononuclear ◽  
High Affinity ◽  
Beta Interferon ◽  
Targeted Mutation ◽  
High Affinity Receptor ◽  
Affinity Receptor ◽  
Alpha Beta

Attenuated Edmonston measles virus (MV-Edm) is not pathogenic in standard mice. We show here that MV-Edm inoculated via the natural respiratory route has a limited propagation in the lungs of mice with a targeted mutation inactivating the alpha/beta interferon receptor. A high dose of MV-Edm administered intracerebrally is lethal for about half of these mice. To study the consequences of the availability of a high-affinity receptor for MV propagation, we generated alpha/beta interferon-defective mice expressing human CD46 with human-like tissue specificity. Intranasal infection of these mice with MV-Edm resulted in enhanced spread to the lungs and more prominent inflammatory response. Virus replication was also detected in peripheral blood mononuclear cells, the spleen, and the liver. Moreover, intracerebral inoculation of adult animals with low MV-Edm doses caused encephalitis with almost inevitably lethal outcome. We conclude that in mice alpha/beta interferon controls MV infection and that a high-affinity receptor facilitates, but is not strictly required for, MV spread and pathogenesis.

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