Comparative analysis of lipotoxicity induced by endocrine, pharmacological, and innate immune stimuli in rat basophilic leukemia cells

Maaetoft-Udsen, Kristina; Greineisen, William E.; Aldan, Johnny Tudela; Magaoay, Hazelle; Ligohr, Cheryll; Shimoda, Lori M. N.; Sung, Carl; Turner, Helen

doi:10.3109/1547691x.2014.990655

Cited by 5 publications

(4 citation statements)

References 33 publications

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“…Glucagon and TNF alpha were used at 10 µM and 20 ng/ml respectively. Lipogenesis was confirmed by LipidTOX (Invitrogen, Temecula, CA) staining as described [ 6 , 21 ].…”

Section: Methodsmentioning

confidence: 99%

Transcriptional and Functional Plasticity Induced by Chronic Insulin Exposure in a Mast Cell-Like Basophilic Leukemia Cell Model

et al. 2017

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ObjectiveSecretory granules (SG) and lipid bodies (LB) are the primary organelles that mediate functional responses in mast cells. SG contains histamine and matrix-active proteases, while LB are reservoirs of arachidonic acid and its metabolites, precursors for rapid synthesis of eicosanoids such as LTC4. Both of these compartments can be dynamically or ontologically regulated, with metabolic and immunological stimuli altering lipid body content and granule numbers responding to contextual signals from tissue. We previously described that chronic in vitro or in vivo hyperinsulinemia expands the LB compartment with a concomitant loss of SG capacity, suggesting that this ratio is dynamically regulated. The objective of the current study is to determine if chronic insulin exposure initiates a transcriptional program that biases model mast cells towards a lipogenic state with accompanying loss of secretory granule biogenesis.MethodsWe used a basophilic leukemic cell line with mucosal mast cell-like features as a model system. We tested the hypothesis that chronic insulin exposure initiates a transcriptional program that biases these model mast cells towards a lipogenic state with accompanying loss of secretory granule biogenesis. Transcriptional arrays were used to map gene expression patterns. Biochemical, immunocytochemical and mediator release assays were used to evaluate organelle numbers and functional responses.ResultsIn a mucosal mast cell model, the rat basophilic leukemia line RBL2H3, mast cell granularity and SG numbers are inversely correlated with LB numbers. Chronic insulin exposure appears to modulate gene networks involved in both lipid body biogenesis and secretory granule formation. Western blot analysis confirms upregulation of protein levels for LB proteins, and decreases in proteins that are markers for SG cargo.ConclusionsThe levels of insulin in the extracellular milieu may modify the phenotype of mast cell-like cells in vitro.

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“…Glucagon and TNF alpha were used at 10 µM and 20 ng/ml respectively. Lipogenesis was confirmed by LipidTOX (Invitrogen, Temecula, CA) staining as described [ 6 , 21 ].…”

Section: Methodsmentioning

confidence: 99%

Transcriptional and Functional Plasticity Induced by Chronic Insulin Exposure in a Mast Cell-Like Basophilic Leukemia Cell Model

et al. 2017

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“…After washing with PBS-T, membranes were incubated with secondary antibody conjugated to horseradish-peroxidase for 60 minutes before being washed with PBS-T, developed with enhanced chemiluminescence (Amersham, Piscataway, NJ) and exposed to hyperfilm-enhanced chemiluminescence (Amersham, Piscataway, NJ). Rat basophilic leukemia (RBL) cells (ATCC, Manassas, VA) were used as a positive control for TLR expression in Western blots [21,22]. …”

Section: Methodsmentioning

confidence: 99%

Human amnion mesenchymal cells are pro-inflammatory when activated by the Toll-like receptor 2/6 ligand, macrophage-activating lipoprotein-2

et al. 2016

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Introduction Infection accounts for over 40% of preterm premature rupture of the fetal membranes (PPROM), a major cause of preterm birth. Toll-like receptors (TLR) play key roles in pathogen surveillance but their expression and function in amnion mesenchymal cells (AMC) is unclear. The aims of this study were to determine the expression of all TLR isoforms and the effect of macrophage-activating lipoprotein-2 (MALP-2), derived from a common pathogen involved in PPROM, on human AMC. Methods AMC were isolated from normal, term, amnion from repeat caesarean section. Semi-quantitative RT-PCR, immunocytochemistry, immunohistochemistry and western blotting were used to detect TLR isoform expression. Immunocytochemistry of NF-κB p65, pro-inflammatory cytokine secretion (ELISA), MTT assay, LDH assay, immunoblotting of cytosolic cytochrome c and cleaved caspase-3, and expression of 84 microRNAs by Qiagen miRNA PCR array were used to determine the functional effect of MALP-2 on AMC. Results TLR1-10 was detected in AMC, and protein expression of TLR2, 4, and 6 were confirmed. MALP-2 induced nuclear translocation of p65, reaching significance after 45 minutes (ANOVA, P < 0.05). MALP-2 did not cause apoptosis but did lead to significant secretion of IL-4, IL-6, and IL-8 (P < 0.05, 0.01, 0.001, respectively) and significant changes in miRNA-320a and miRNA-18a (P < 0.05). Discussion These results suggest that AMC elicit a pro-inflammatory response following stimulation with the known TLR2/6 ligand MALP-2. This data supports the idea that AMC express the innate immune system receptors that could help with immune surveillance during infection and contribute to inflammatory responses that lead to PPROM.

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“…Adipocytes, the primary cells forming adipose tissue, contain cytosolic lipid droplets composed of a core of neutral lipids, including sterol esters and TGs, surrounded by a phospholipid monolayer [ 218 , 219 , 220 , 221 , 222 , 223 , 224 ]. Lipotoxicity is the abnormal accumulation of lipid droplets in non-adipose tissues, which leads to mitochondrial dysfunction, inhibition of ATP generation, and ultimately cell death by apoptosis [ 217 , 225 , 226 ]. Manifestation of lipotoxicity typically occurs in kidney, liver, heart and skeletal muscle [ 217 ].…”

Section: Lipid Biomarkers For Non-infectious Diseasesmentioning

confidence: 99%

Detection of Lipid and Amphiphilic Biomarkers for Disease Diagnostics

Kubicek-Sutherland

Mendez

et al. 2017

Biosensors

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Rapid diagnosis is crucial to effectively treating any disease. Biological markers, or biomarkers, have been widely used to diagnose a variety of infectious and non-infectious diseases. The detection of biomarkers in patient samples can also provide valuable information regarding progression and prognosis. Interestingly, many such biomarkers are composed of lipids, and are amphiphilic in biochemistry, which leads them to be often sequestered by host carriers. Such sequestration enhances the difficulty of developing sensitive and accurate sensors for these targets. Many of the physiologically relevant molecules involved in pathogenesis and disease are indeed amphiphilic. This chemical property is likely essential for their biological function, but also makes them challenging to detect and quantify in vitro. In order to understand pathogenesis and disease progression while developing effective diagnostics, it is important to account for the biochemistry of lipid and amphiphilic biomarkers when creating novel techniques for the quantitative measurement of these targets. Here, we review techniques and methods used to detect lipid and amphiphilic biomarkers associated with disease, as well as their feasibility for use as diagnostic targets, highlighting the significance of their biochemical properties in the design and execution of laboratory and diagnostic strategies. The biochemistry of biological molecules is clearly relevant to their physiological function, and calling out the need for consideration of this feature in their study, and use as vaccine, diagnostic and therapeutic targets is the overarching motivation for this review.

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Comparative analysis of lipotoxicity induced by endocrine, pharmacological, and innate immune stimuli in rat basophilic leukemia cells

Cited by 5 publications

References 33 publications

Transcriptional and Functional Plasticity Induced by Chronic Insulin Exposure in a Mast Cell-Like Basophilic Leukemia Cell Model

Transcriptional and Functional Plasticity Induced by Chronic Insulin Exposure in a Mast Cell-Like Basophilic Leukemia Cell Model

Human amnion mesenchymal cells are pro-inflammatory when activated by the Toll-like receptor 2/6 ligand, macrophage-activating lipoprotein-2

Detection of Lipid and Amphiphilic Biomarkers for Disease Diagnostics

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