scholarly journals RNA-Seq Analysis of Activated PBMC Treated With LMWF5A Predicts an Anti-Inflammatory Mode of Action and Similar Drug Targets to Dexamethasone

2021 ◽  
Author(s):  
Elizabeth D. Frederick ◽  
Melissa A. Hausburg ◽  
Gregory W. Thomas ◽  
David Bar-Or
Keyword(s):  
Drug Targets ◽  
Mononuclear Cells ◽  
Biological Effects ◽  
Weight Fraction ◽  
Interferon Γ ◽  
Immunomodulatory Activity ◽  
Peripheral Blood Mononuclear ◽  
Inflammatory Conditions ◽  
Anti Inflammatory ◽  
Upstream Regulators

Abstract Background: The low molecular weight fraction of human serum albumin (LMWF5A) has immunomodulatory activity via its effects on multiple inflammatory mediators and is currently being evaluated for the treatment of hyperactive or persistent inflammatory conditions. To gain further insight into the mechanism of action (MOA) of LMWF5A, an investigation of its effects on activated immune cells was performed. Methods and Results: Peripheral blood mononuclear cells (PBMC) were treated with vehicle control or LMWF5A and stimulated with lipopolysaccharide (LPS), LPS/interferon γ, or interleukin (IL)-4/IL-13, and RNAseq was performed to determine differentially expressed genes (DEGs) within each condition. Unbiased Ingenuity Pathway Analysis (IPA) of DEGs revealed anti-inflammatory and pro-resolving activities for LMWF5A. Moreover, comparison to all IPA upstream regulators predicted that the LMWF5A MOA is opposite to pro-inflammatory regulators and significantly matches the activity of several anti-inflammatory molecules. These analyses identified the glucocorticoid dexamethasone (DEX) as the most significantly similar regulator to LMWF5A. To further explore similarities to DEX, LMWF5A DEGs were compared to two publicly available datasets of activated, DEX-treated PBMC. These comparisons showed continuity between predicted upstream regulators, affording further support to the hypothesis that LMWF5A acts in a manner like DEX. Nevertheless, not all LMWF5A-targeted DEGs showed directional regulation identical to DEX. Conclusions: This study further defines the MOA of LMWF5A and provides hypotheses for future investigations. Because of its predicted similar biological effects and known safety profile, LMWF5A could potentially be used to treat conditions that are supported for DEX with fewer or less harmful side effects.

scholarly journals LMWF5A Demonstrates an Anti-Inflammatory Mode of Action and Similar Drug Targets to Dexamethasone in Activated PBMC

2021 ◽  
Author(s):  
Elizabeth D. Frederick ◽  
Melissa A. Hausburg ◽  
Gregory W. Thomas ◽  
David Bar-Or
Keyword(s):  
Inflammatory Mediators ◽  
Mode Of Action ◽  
Drug Targets ◽  
Mononuclear Cells ◽  
Current Knowledge ◽  
Biological Effects ◽  
Weight Fraction ◽  
Immunomodulatory Activity ◽  
Peripheral Blood Mononuclear ◽  
Anti Inflammatory

Abstract The low molecular weight fraction of human serum albumin (LMWF5A) has proven immunomodulatory activity via its effects on multiple inflammatory mediators and transcription factors. This biologic drug is currently being evaluated for the treatment of conditions characterized by hyperactive or persistent immune responses. To gain further insight into the mechanism of action of LMWF5A, an unbiased investigation of its effects on activated immune cells was performed. Peripheral blood mononuclear cells (PBMC) were treated with vehicle control or LMWF5A and stimulated with lipopolysaccharide (LPS), LPS/interferon g, or interleukin (IL)-4/IL-13 for 24h. Differential gene expression analysis by RNAseq was performed to determine transcriptional changes due to LMWF5A treatment in each stimulation condition. Ingenuity Pathway Analysis software was employed to reveal overall biological trends observed in the RNAseq datasets and to identify compounds with comparable or contrasting activities to LMWF5A. Analysis of each RNA dataset revealed broad anti-inflammatory and pro-resolving activities for LMWF5A, with predicted inhibition of key pro-inflammatory mediators and functions as well as activation of some anti-inflammatory mediators. Upstream regulator analysis confirmed these activities and identified the corticosteroid dexamethasone as the most significantly similar regulator. Further comparison of LMWF5A and dexamethasone revealed similar, but not identical, targets and directional regulation. This study supports our current knowledge on the mode of action of LMWF5A and provides new hypotheses for future investigations. Due to its analogous biological effects with dexamethasone and known safety profile, LMWF5A may be used to treat conditions that are supported for dexamethasone with fewer or less harmful side effects.

scholarly journals Anti-inflammatory genes in PBMCs post-spontaneous intracerebral hemorrhage

2021 ◽  
Vol 12 (1) ◽  
pp. 58-66
Author(s):  
Doan Nguyen ◽  
Vi Tran ◽  
Alireza Shirazian ◽  
Cruz Velasco-Gonzalez ◽  
Ifeanyi Iwuchukwu
Keyword(s):  
Intracerebral Hemorrhage ◽  
Negative Correlation ◽  
Mononuclear Cells ◽  
Inflammatory Cells ◽  
Favorable Outcome ◽  
Hospital Length Of Stay ◽  
Spontaneous Intracerebral Hemorrhage ◽  
Hematoma Volume ◽  
Peripheral Blood Mononuclear ◽  
Anti Inflammatory

Abstract Background Neuroinflammation is important in the pathophysiology of spontaneous intracerebral hemorrhage (ICH) and peripheral inflammatory cells play a role in the clinical evolution and outcome. Methodology Blood samples from ICH patients (n = 20) were collected at admission for 5 consecutive days for peripheral blood mononuclear cells (PBMCs). Frozen PBMCs were used for real-time PCR using Taqman probes (NFKB1, SOD1, PPARG, IL10, NFE2L2, and REL) and normalized to GAPDH. Data on hospital length of stay and modified Rankin score (MRS) were collected with 90-day MRS ≤ 3 as favorable outcome. Statistical analysis of clinical characteristics to temporal gene expression from early to delayed timepoints was compared for MRS groups (favorable vs unfavorable) and hematoma volume. Principle findings and results IL10, SOD1, and REL expression were significantly higher at delayed timepoints in PBMCs of ICH patients with favorable outcome. PPARG and REL increased between timepoints in patients with favorable outcome. NFKB1 expression was not sustained, but significantly decreased from higher levels at early onset in patients with unfavorable outcome. IL10 expression showed a negative correlation in patients with high hematoma volume (>30 mL). Conclusions and significance Anti-inflammatory, pro-survival regulators were highly expressed at delayed time points in ICH patients with a favorable outcome, and IL10 expression showed a negative correlation to high hematoma volume.

scholarly journals In Vitro Anti-Inflammatory and Immunomodulatory Activities of an Extract from the Roots of Bupleurum rotundifolium

Medicines ◽  
2019 ◽  
Vol 6 (4) ◽  
pp. 101 ◽  
Author(s):  
Cholet ◽  
Decombat ◽  
Vareille-Delarbre ◽  
Gainche ◽  
Berry ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Polymorphonuclear Neutrophils ◽  
Ros Production ◽  
Peripheral Blood Mononuclear ◽  
Aerial Parts ◽  
Bupleurum Scorzonerifolium Willd ◽  
Anti Inflammatory ◽  
Inflammatory Profile ◽  
Bupleurum Scorzonerifolium

Background: Some Bupleurum species, such as the Bupleurum chinense DC. or the Bupleurum scorzonerifolium Willd have been extensively studied (especially their roots) for the treatment of inflammation. In contrast, only compounds extracted from the aerial parts of Bupleurum rotundifolium have been studied and showed anti-inflammatory or antiproliferative activities. This study was conducted to investigate the antioxidant, anti-inflammatory, and immunomodulatory effects of Bupleurum rotundifolium roots. Methods: To tackle the various aspects of inflammation, we studied in vitro a methanolic extract from the roots of Bupleurum rotundifolium on peripheral blood mononuclear cells (PBMCs), polymorphonuclear neutrophils (PMNs), and the monocytic cells THP-1. Its antioxidant capacities and iron-chelating activity were assessed. The extract was tested on THP-1 differentiation, reactive oxygen species (ROS) production by leukocytes, neutrophils chemotaxis, cytokines, PGE2 production, and NF-κB activation in PBMCs. Results: The extract showed a decreased ROS production in stimulated cells. It increased PBMC chemokine secretion and up-regulated the differentiation of THP-1 monocytes into macrophage-like cells, indicating a potential interest of the extract in the resolution of acute inflammation. In addition, the analysis of cytokine production suggests that Bupleurum rotundifolium has immunomodulatory properties. Conclusions: Cytokines secretion, especially IL-1β and IL-12p70, provided us with a set of indicators suggesting that the extract might be able to drive the polarization of macrophages and lymphocytes toward a Th2 anti-inflammatory profile in excessive inflammation.

scholarly journals Phytochemical characterization and biological activities of Plectranthus barbatus Andrews

2022 ◽  
Vol 82 ◽  
Author(s):  
M. F. Cordeiro ◽  
T. R. S. Nunes ◽  
F. G. Bezerra ◽  
P. K. M. Damasco ◽  
W. A. V. Silva ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Trichophyton Rubrum ◽  
Biological Activities ◽  
Sandwich Elisa ◽  
Immunomodulatory Activity ◽  
Potential Candidate ◽  
Leaf Extracts ◽  
Healthy Human ◽  
Peripheral Blood Mononuclear ◽  
Normal Peripheral Blood

Abstract Plectranthus barbatus Andrews (Lamiaceae) is widely distributed in the world and has a range of popular therapeutic indications. This work aimed to evaluate the phytochemical characterization of two leaf extracts of P. barbatus, and their antimicrobial, antineoplastic and immunomodulatory potential. After collection, herborization and obtainment of the P. barbatus aqueous extract (PBA) and acetone:water 7:3 P. barbatus organic extract (PBO), the phytochemical characterization was performed by high-performance liquid chromatography (HPLC). The antimicrobial activity was performed to determine the minimum inhibitory concentration (MIC) against eight bacterial strains using the microdilution test and the fungus Trichophyton rubrum by disc diffusion assay and microdilution test. Cytotoxicity was assessed by MTT and trypan blue methods in normal peripheral blood mononuclear cells (PBMCs) at concentrations ranged between 0.1 to 100 µg.mL-1 and in neoplastic cell lines Toledo, K562, DU-145 and PANC-1 at 1, 10 and 100 µg.mL-1 . Immunomodulatory activity, was evaluated by sandwich ELISA of proinflammatory cytokines at BALB/c mice splenocytes cultures supernatant. Both extracts presented flavonoids, cinnamic derivatives, steroids and ellagic acid. PBO showed bacteriostatic activity against Acinetobacter baumannii (MIC = 250 µg.mL-1) clinical isolate and PBA fungistatic activity against Trichophyton rubrum (MIC = 800 µg.mL-1). The extracts did not exhibit toxicity to PBMCs and neoplastic cells (IC50 > 100 µg.mL-1). Additionally, PBO at 100 µg.mL-1 significantly inhibited IFN-γ and IL-17A cytokines (p = 0.03). Plectranthus barbatus is a potential candidate for therapeutic use due to its low toxicity in healthy human cells and exhibits biological activities of medical interest as bacteriostatic, fungistatic and immunomodulatory.

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