LPS-induced bronchial hyperreactivity: interference by mIL-10 differs according to site of delivery

When administered to mice systemically or via the airways, LPS induces bronchoconstriction (BC) and/or bronchopulmonary hyperreactivity (BHR), associated with inflammation. Accordingly, a relationship between inflammation and allergic and nonallergic BHR can be hypothesized. We therefore studied the interference of the anti-inflammatory cytokine murine IL-10 (mIL-10) with LPS-induced lung inflammation, BC, and BHR. mIL-10 was administered directly into the airways by intranasal instillation or generated in vivo after muscle electrotransfer of mIL-10-encoding plasmid. Electrotransfer led to high mIL-10 circulating levels for a longer time than after the injection of recombinant mIL-10 (rmIL-10). rmIL-10 administered intranasally reduced lung inflammation and BHR after LPS administration into airways. It also reduced the ex vivo production of TNF-α by LPS-stimulated lung tissue explants. Two days after electrotransfer, mIL-10 blood levels were elevated, but lung inflammation, BC, and BHR persisted unaffected. Blood mIL-10 reaches the airways poorly, which probably accounts for the ineffectiveness of mIL-10-encoding plasmid electrotransfer. When LPS was aerosolized 15 days after electrotransfer, lung inflammation persisted but BHR was significantly reduced, an effect that may be related to the longer exposure of the relevant cells to mIL-10. The dissociation between inflammation and BHR indicates that both are not directly correlated. In conclusion, this study shows that mIL-10 is efficient against BHR when present in the airway compartment. Despite this, the muscle electrotransfer with mIL-10-encoding plasmid showed a protective effect against BHR after a delay of 2 wk that should be further investigated.

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Hyperthermia increases interleukin-6 in mouse skeletal muscle

AJP Cell Physiology ◽

10.1152/ajpcell.00028.2012 ◽

2012 ◽

Vol 303 (4) ◽

pp. C455-C466 ◽

Cited By ~ 49

Author(s):

Steven S. Welc ◽

Neil A. Phillips ◽

Jose Oca-Cossio ◽

Shannon M. Wallet ◽

Daniel L. Chen ◽

...

Keyword(s):

Gene Expression ◽

Skeletal Muscle ◽

Core Temperature ◽

Ex Vivo ◽

C2c12 Myotubes ◽

Tnf Α ◽

Passive Hyperthermia ◽

Circulating Levels ◽

Cell Supernatant

Skeletal muscles produce and contribute to circulating levels of IL-6 during exercise. However, when core temperature is reduced, the response is attenuated. Therefore, we hypothesized that hyperthermia may be an important and independent stimulus for muscle IL-6. In cultured C2C12 myotubes, hyperthermia (42°C) increased IL-6 gene expression 14-fold after 1 h and 35-fold after 5 h of 37°C recovery; whereas exposure to 41°C resulted in a 2.6-fold elevation at 1 h. IL-6 protein was secreted and significantly elevated in the cell supernatant. Similar but reduced responses to heat were seen in C2C12 myoblasts. Isolated soleus muscles from mice, exposed ex vivo to 41°C for 1 h, yielded similar IL-6 gene responses (>3-fold) but without a significant effect on protein release. When whole animals were exposed to passive hyperthermia, such that core temperature increased to 42.4°C, IL-6 mRNA in soleus increased 5.4-fold compared with time matched controls. Interestingly, TNF-α gene expression was routinely suppressed at all levels of hyperthermia (40.5–42°C) in the isolated models, but TNF-α was elevated (4.2-fold) in the soleus taken from intact mice exposed, in vivo, to hyperthermia. Muscle HSP72 mRNA increased as a function of the level of hyperthermia, and IL-6 mRNA responses increased proportionally with HSP72. In cultured C2C12 myotubes, when heat shock factor was pharmacologically blocked with KNK437, both HSP72 and IL-6 mRNA elevations, induced by heat, were suppressed. These findings implicate skeletal muscle as a “heat stress sensor” at physiologically relevant hyperthermia, responding with a programmed cytokine expression pattern characterized by elevated IL-6.

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Decellularised Human Umbilical Artery as a Vascular Graft Elicits Minimal Pro-Inflammatory Host Response Ex Vivo and In Vivo

International Journal of Molecular Sciences ◽

10.3390/ijms22157981 ◽

2021 ◽

Vol 22 (15) ◽

pp. 7981

Author(s):

Alexander Høgsted Ahlmann ◽

Shu Fang ◽

Sussi Bagge Mortensen ◽

Line Weis Andersen ◽

Pernille Gejl Pedersen ◽

...

Keyword(s):

Vascular Graft ◽

Umbilical Artery ◽

Ex Vivo ◽

White Blood Cells ◽

Small Diameter ◽

M1 Macrophages ◽

Host Immune Responses ◽

Cytokine Levels ◽

Anti Inflammatory Cytokine

Small diameter (<6 mm) vessel grafts still pose a challenge for scientists worldwide. Decellularised umbilical artery (dUA) remains promising as small diameter tissue engineered vascular graft (TEVG), yet their immunogenicity remains unknown. Herein, we evaluated the host immune responses, with a focus on the innate part, towards human dUA implantation in mice, and confirmed our findings in an ex vivo allogeneic human setup. Overall, we did not observe any differences in the number of circulating white blood cells nor the number of monocytes among three groups of mice (1) dUA patch; (2) Sham; and (3) Mock throughout the study (day −7 to 28). Likewise, we found no difference in systemic inflammatory and anti-inflammatory cytokine levels between groups. However, a massive local remodelling response with M2 macrophages were observed in the dUA at day 28, whereas M1 macrophages were less frequent. Moreover, human monocytes from allogeneic individuals were differentiated into macrophages and exposed to lyophilised dUA to maximize an eventual M1 response. Yet, dUA did not elicit any immediate M1 response as determined by the absence of CCR7 and CXCL10. Together this suggests that human dUA elicits a minimal pro-inflammatory response further supporting its use as a TEVG in an allogeneic setup.

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Combining Immunocytokine and Ex Vivo Activated NK Cells as a Platform for Enhancing Graft-Versus-Tumor Effects Against GD2+ Murine Neuroblastoma

Frontiers in Immunology ◽

10.3389/fimmu.2021.668307 ◽

2021 ◽

Vol 12 ◽

Author(s):

Paul D. Bates ◽

Alexander L. Rakhmilevich ◽

Monica M. Cho ◽

Myriam N. Bouchlaka ◽

Seema L. Rao ◽

...

Keyword(s):

Tumor Growth ◽

Nk Cells ◽

Nk Cell ◽

Ex Vivo ◽

Cell Transplant ◽

Hematopoietic Stem ◽

Allogeneic Hsct ◽

Tnf Α

Management for high-risk neuroblastoma (NBL) has included autologous hematopoietic stem cell transplant (HSCT) and anti-GD2 immunotherapy, but survival remains around 50%. The aim of this study was to determine if allogeneic HSCT could serve as a platform for inducing a graft-versus-tumor (GVT) effect against NBL with combination immunocytokine and NK cells in a murine model. Lethally irradiated C57BL/6 (B6) x A/J recipients were transplanted with B6 bone marrow on Day +0. On day +10, allogeneic HSCT recipients were challenged with NXS2, a GD2+ NBL. On days +14-16, mice were treated with the anti-GD2 immunocytokine hu14.18-IL2. In select groups, hu14.18-IL2 was combined with infusions of B6 NK cells activated with IL-15/IL-15Rα and CD137L ex vivo. Allogeneic HSCT alone was insufficient to control NXS2 tumor growth, but the addition of hu14.18-IL2 controlled tumor growth and improved survival. Adoptive transfer of ex vivo CD137L/IL-15/IL-15Rα activated NK cells with or without hu14.18-IL2 exacerbated lethality. CD137L/IL-15/IL-15Rα activated NK cells showed enhanced cytotoxicity and produced high levels of TNF-α in vitro, but induced cytokine release syndrome (CRS) in vivo. Infusing Perforin-/- CD137L/IL-15/IL-15Rα activated NK cells had no impact on GVT, whereas TNF-α-/- CD137L/IL-15/IL-15Rα activated NK cells improved GVT by decreasing peripheral effector cell subsets while preserving tumor-infiltrating lymphocytes. Depletion of Ly49H+ NK cells also improved GVT. Using allogeneic HSCT for NBL is a viable platform for immunocytokines and ex vivo activated NK cell infusions, but must be balanced with induction of CRS. Regulation of TNFα or activating NK subsets may be needed to improve GVT effects.

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Effect of the proinflammatory cytokine TNF-α on intestinal riboflavin uptake: inhibition mediated via transcriptional mechanism(s)

AJP Cell Physiology ◽

10.1152/ajpcell.00295.2018 ◽

2018 ◽

Vol 315 (5) ◽

pp. C653-C663 ◽

Cited By ~ 5

Author(s):

Kasin Yadunandam Anandam ◽

Omar A. Alwan ◽

Veedamali S. Subramanian ◽

Padmanabhan Srinivasan ◽

Rubina Kapadia ◽

...

Keyword(s):

Ex Vivo ◽

Significant Inhibition ◽

Mrna Levels ◽

In Vivo Models ◽

Uptake Inhibition ◽

Tnf Α ◽

Vitro Model ◽

Factor Α

Riboflavin (RF), is essential for normal cellular metabolism/function. Intestinal RF absorption occurs via a specific carrier-mediated process that involves the apical transporter RFVT-3 ( SLC52A3) and the basolateral RFVT-1 (SLC52A1). Previously, we characterized different cellular/molecular aspects of the intestinal RF uptake process, but nothing is known about the effect of proinflammatory cytokines on the uptake event. We addressed this issue using in vitro, ex vivo, and in vivo models. First, we determined the level of mRNA expression of the human (h)RFVT-3 and hRFVT-1 in intestinal tissue of patients with inflammatory bowel disease (IBD) and observed a markedly lower level compared with controls. In the in vitro model, exposing Caco-2 cells to tumor necrosis factor-α (TNF-α) led to a significant inhibition in RF uptake, an effect that was abrogated upon knocking down TNF receptor 1 (TNFR1). The inhibition in RF uptake was associated with a significant reduction in the expression of hRFVT-3 and -1 protein and mRNA levels, as well as in the activity of the SLC52A3 and SLC52A1 promoters. The latter effects appear to involve Sp1 and NF-κB sites in these promoters. Similarly, exposure of mouse small intestinal enteroids and wild-type mice to TNF-α led to a significant inhibition in physiological and molecular parameters of intestinal RF uptake. Collectively, these findings demonstrate that exposure of intestinal epithelial cells to TNF-α leads to inhibition in RF uptake and that this effect is mediated, at least in part, via transcriptional mechanism(s). These findings may explain the significantly low RF levels observed in patients with IBD.

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P0527AROLE OF PROMOTING INFLAMMATION OF KLF6 IN ACUTE KIDNEY INURY

Nephrology Dialysis Transplantation ◽

10.1093/ndt/gfaa142.p0527a ◽

2020 ◽

Vol 35 (Supplement_3) ◽

Author(s):

Dan Li ◽

Chenyu Li ◽

Yan Xu

Keyword(s):

Inflammatory Cytokines ◽

Inflammatory Cytokine ◽

Ischemia Reperfusion ◽

Kidney Injury ◽

Tnf Α ◽

Public Dataset ◽

Anti Inflammatory ◽

Anti Inflammatory Cytokine ◽

Pro Inflammatory Cytokines

Abstract Background and Aims Acute kidney injury (AKI), commonly appeared in cardiac arrest, surgery and kidney transplantation which involved in ischemia-reperfusion (IR) injury of kidney. However, the mechanisms underlying inflammatory response in IR AKI is still unclear. Method Public dataset showed kruppel-like factor 6 (KLF6) was significantly highly expressed (P<0.05) in AKI, implies KLF6 might be associated with AKI. To evaluate the mechanism of KLF6 on IR AKI, 30 rats were randomly divided into sham and IR group, and were sacrificed at 0 h, 3 h, 6 h, 12 h or 24 h after IR. Results The results showed KLF6 expression was peaking at 6 h after IR, and the expression of pro-inflammatory cytokines MCP-1 and TNF-α were increased both in serum and kidney tissues after IR, while anti-inflammatory cytokine IL-10 was decreased after IR. Furthermore, in vitro results showed KLF6 knock-down reduced the pro-inflammatory cytokines expression and increased the anti-inflammatory cytokines expression. Conclusion These results suggest that (1) KLF6 might be a novel biomarker for early diagnosis of AKI and (2) targeting KLF6 expression may offer novel strategies to protect kidneys from IR AKI Figure KLF6, AKI, Control Inflammation

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PROTECTIVE EFFECT OF A NEW SYNTHETIC COMPOUND: PCA-4230, ON SEVERAL In vivo THROMBOSIS MODELS.

10.1055/s-0038-1643430 ◽

1987 ◽

Author(s):

M P Ortega ◽

C Sunkel ◽

J G Priego

Keyword(s):

Protective Effect ◽

Bleeding Time ◽

Ex Vivo ◽

Platelet Activating Factor ◽

Phase I Trials ◽

Synthetic Compound ◽

Prolonged Bleeding Time ◽

Series Of Experiments ◽

Pre Treatment

PCA-4230 is a new anti-thrombotic compound which inhibits pla. telet aggregation In vltn.0 and ex. vivo in several species, including man, prolongs the bleeding time and has potent protective ac tivity in several thrombosis models. Phase I trials with different dosage schedules have recently been initiated.In the present study, the effects of PCA-4230 on bleeding time and on several In vivo thrombosis models were studied in mice. Mice were treated with one single oral dose of PCA-4230 (1-10 mg/ kg). One hour after treatment, mice were injected intravenously with four thrombotic challengers {arachidonic acid (AA), thromboxane agonist (U46619), Platelet Activating Factor (PAF) or collagen/epinephrine combination (C/E)} at a dose which induced 80-90% of mortality. The thrombotic agents were prepared in saline. The appropiate doses were dissolved in a volume of 100 μl/mouse. Bleeding time was measured in non-anesthetized mice by the tail transection technique.Effects of compound were recorded from1 to 4 hours after dosage. Acute pre-treatment with PCA-4230 showed a significant dose-depen dent protective effect.Results of each series of experiments are given in the next table.The compound inhibited thrombotic sudden death induced by U46619, PAF or C/E combination, reduced the duration of respiratory distress induced by AA and prolonged bleeding time. Thus, PCA-4230 is protective against a variety of thrombotic stimuli.These results suggest that PCA-4230 may be a promising anti-throm botic drug.

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Inhibition of endotoxin-induced lung inflammation by interleukin-10 gene transfer in mice

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.2000.279.5.l872 ◽

2000 ◽

Vol 279 (5) ◽

pp. L872-L877 ◽

Cited By ~ 11

Author(s):

Sujatha Dokka ◽

Carl J. Malanga ◽

Xianglin Shi ◽

Fei Chen ◽

Vincent Castranova ◽

...

Keyword(s):

Gene Transfer ◽

Lung Inflammation ◽

Transgene Expression ◽

Tumor Necrosis ◽

Interleukin 10 ◽

Inhibitory Effect ◽

Biologically Active ◽

Tnf Α ◽

Anti Inflammatory Cytokine ◽

Necrosis Factor

Interleukin (IL)-10 is an anti-inflammatory cytokine that has great potential for use in the treatment of inflammatory and immune illnesses. In this study, gene transfer was used to induce IL-10 transgene expression in murine lungs for treatment of endotoxin-induced lung inflammation. Gene transfer was performed with a cytomegalovirus (CMV)-IL-10 plasmid with the aid of the liposomal agents LipofectAMINE and N-[1-(2,3-dioleoyl)propyl]- N, N, N-trimethylammonium methylsulfate (DOTAP). Administration of the endotoxin caused a marked increase in lung inflammation as indicated by increased tumor necrosis factor (TNF)-α release and neutrophil count. Pretreatment of the mice with IL-10 plasmid with and without LipofectAMINE had no inhibitory effect on lung inflammation and IL-10 transgene expression. LipofectAMINE by itself induced lung inflammation, an effect that was not observed with DOTAP. IL-10 plasmid when codelivered with DOTAP expressed biologically active IL-10 protein and caused a reduction in endotoxin-induced inflammation. Transgene expression was observed as early as 3 h after administration, peaked at 12 h, and declined thereafter. We conclude that IL-10 gene transfer is a feasible approach for the treatment of lung inflammation.

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Rupintrivir reduces RV-induced TH-2 cytokine IL-4 in precision-cut lung slices (PCLS) of HDM-sensitized mice ex vivo

Respiratory Research ◽

10.1186/s12931-019-1175-y ◽

2019 ◽

Vol 20 (1) ◽

Author(s):

Olga Danov ◽

Lisa Lasswitz ◽

Helena Obernolte ◽

Christina Hesse ◽

Armin Braun ◽

...

Keyword(s):

Ex Vivo ◽

Antiviral Drug ◽

Cytokine Response ◽

Experimental Conditions ◽

Tnf Α ◽

Dust Mite ◽

Ifn Γ ◽

Inflammatory Immune Response ◽

Precision Cut Lung Slices

Abstract Background Antiviral drugs such as rupintrivir may have an immune-modulatory effect in experimentally induced allergic asthma with subsequent RV infection. We infected lung slices of house-dust mite (HDM)-sensitized asthmatic mice ex vivo with human rhinovirus (RV) and investigated the effect of the antiviral drug rupintrivir on RV-induced cytokine response in lung tissue of HDM-sensitized mice ex vivo. Methods Mice were sensitized with HDM. Precision-cut lung slices (PCLS) were prepared from HDM-sensitized or non-sensitized mice. Lung slices were infected ex vivo with RV or RV together with rupintrivir. Modulation of immune responses was evaluated by cytokine secretion 48 h post infection. Results In vivo HDM sensitization resulted in a TH-2/TH-17-dominated cytokine response that persisted in PCLS ex vivo. RV infection of PCLS from non-sensitized mice resulted in the induction of an antiviral and pro-inflammatory immune response, as indicated by the secretion of IFN-α, IFN-β, IFN-γ, TNF-α, MCP-1, IP-10, IL-10, and IL-17A. In contrast, PCLS from HDM-sensitized mice showed an attenuated antiviral response, but exaggerated IL-4, IL-6, and IL-10 secretion upon infection. Rupintrivir inhibited exaggerated pro-inflammatory cytokine IL-6 and TH-2 cytokine IL-4 in HDM-sensitized mice. Conclusions In summary, this study demonstrates that treatment with rupintrivir influences virus-induced IL-4 and IL-6 cytokine release under experimental conditions ex vivo.

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RANTES release by human adipose tissue in vivo and evidence for depot-specific differences

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.90511.2008 ◽

2009 ◽

Vol 296 (6) ◽

pp. E1262-E1268 ◽

Cited By ~ 42

Author(s):

Rana Madani ◽

Kalypso Karastergiou ◽

Nicola C. Ogston ◽

Nazar Miheisi ◽

Rahul Bhome ◽

...

Keyword(s):

Adipose Tissue ◽

Subcutaneous Adipose Tissue ◽

Epicardial Adipose Tissue ◽

Ex Vivo ◽

Human Adipose Tissue ◽

Fat Pad ◽

Obese Subjects ◽

Subcutaneous Adipose ◽

Circulating Levels

Obesity is associated with elevated inflammatory signals from various adipose tissue depots. This study aimed to evaluate release of regulated on activation, normal T cell expressed and secreted (RANTES) by human adipose tissue in vivo and ex vivo, in reference to monocyte chemoattractant protein-1 (MCP-1) and interleukin-6 (IL-6) release. Arteriovenous differences of RANTES, MCP-1, and IL-6 were studied in vivo across the abdominal subcutaneous adipose tissue in healthy Caucasian subjects with a wide range of adiposity. Systemic levels and ex vivo RANTES release were studied in abdominal subcutaneous, gastric fat pad, and omental adipose tissue from morbidly obese bariatric surgery patients and in thoracic subcutaneous and epicardial adipose tissue from cardiac surgery patients without coronary artery disease. Arteriovenous studies confirmed in vivo RANTES and IL-6 release in adipose tissue of lean and obese subjects and release of MCP-1 in obesity. However, in vivo release of MCP-1 and RANTES, but not IL-6, was lower than circulating levels. Ex vivo release of RANTES was greater from the gastric fat pad compared with omental ( P = 0.01) and subcutaneous ( P = 0.001) tissue. Epicardial adipose tissue released less RANTES than thoracic subcutaneous adipose tissue in lean ( P = 0.04) but not obese subjects. Indexes of obesity correlated with epicardial RANTES but not with systemic RANTES or its release from other depots. In conclusion, RANTES is released by human subcutaneous adipose tissue in vivo and in varying amounts by other depots ex vivo. While it appears unlikely that the adipose organ contributes significantly to circulating levels, local implications of this chemokine deserve further investigation.

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