Age- and disease-related innate immunity of human leukocytes ex vivo

2012 ◽  
Vol 47 (1) ◽  
pp. 8-13 ◽  
Author(s):  
Bogna Jatczak ◽  
Jerzy Leszek ◽  
Iwona Siemieniec ◽  
Marta Sochocka ◽  
Agnieszka Wiśniewska ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Ex Vivo ◽  
Human Leukocytes

Alpha-Amylase/Trypsin Inhibitors, novel activators of innate immunity in ex-vivo tissue explants

2013 ◽  
Vol 51 (08) ◽  
Author(s):  
V Zevallos ◽  
P Olinga ◽  
Y Junker ◽  
PB Tung ◽  
N Volz ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Ex Vivo ◽  
Trypsin Inhibitors ◽  
Alpha Amylase ◽  
Tissue Explants ◽  
Ex Vivo Tissue

scholarly journals Hookworm-Derived Metabolites Suppress Pathology in a Mouse Model of Colitis and Inhibit Secretion of Key Inflammatory Cytokines in Primary Human Leukocytes

2019 ◽  
Vol 87 (4) ◽  
Author(s):  
Phurpa Wangchuk ◽  
Catherine Shepherd ◽  
Constantin Constantinoiu ◽  
Rachael Y. M. Ryan ◽  
Konstantinos A. Kouremenos ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Inflammatory Cytokines ◽  
Mononuclear Cells ◽  
Ex Vivo ◽  
Clinical Signs ◽  
Gas Chromatography Mass Spectrometry ◽  
Trinitrobenzenesulfonic Acid ◽  
Human Leukocytes ◽  
Polar Metabolites ◽  
Anti Inflammatory

ABSTRACT Iatrogenic hookworm therapy shows promise for treating disorders that result from a dysregulated immune system, including inflammatory bowel disease (IBD). Using a murine model of trinitrobenzenesulfonic acid-induced colitis and human peripheral blood mononuclear cells, we demonstrated that low-molecular-weight metabolites derived from both somatic extracts (LMWM-SE) and excretory-secretory products (LMWM-ESP) of the hookworm, Ancylostoma caninum, display anti-inflammatory properties. Administration to mice of LMWM-ESP as well as sequentially extracted fractions of LMWM-SE using both methanol (SE-MeOH) and hexane-dichloromethane-acetonitrile (SE-HDA) resulted in significant protection against T cell-mediated immunopathology, clinical signs of colitis, and impaired histological colon architecture. To assess bioactivity in human cells, we stimulated primary human leukocytes with lipopolysaccharide in the presence of hookworm extracts and showed that SE-HDA suppressed ex vivo production of inflammatory cytokines. Gas chromatography-mass spectrometry (MS) and liquid chromatography-MS analyses revealed the presence of 46 polar metabolites, 22 fatty acids, and five short-chain fatty acids (SCFAs) in the LMWM-SE fraction and 29 polar metabolites, 13 fatty acids, and six SCFAs in the LMWM-ESP fraction. Several of these small metabolites, notably the SCFAs, have been previously reported to have anti-inflammatory properties in various disease settings, including IBD. This is the first report showing that hookworms secrete small molecules with both ex vivo and in vivo anti-inflammatory bioactivity, and this warrants further exploration as a novel approach to the development of anti-inflammatory drugs inspired by coevolution of gut-dwelling hookworms with their vertebrate hosts.

scholarly journals The SARS-CoV-2 spike protein primes inflammasome-mediated interleukin-1- beta secretion in COVID-19 patient-derived macrophages

2020 ◽  
Author(s):  
Sebastian J. Theobald ◽  
Alexander Simonis ◽  
Christoph Kreer ◽  
Matthias Zehner ◽  
Julia Fischer ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Drug Treatment ◽  
Ex Vivo ◽  
Interleukin 1 ◽  
Vaccine Design ◽  
Spike Protein ◽  
Inflammasome Activation ◽  
Interleukin 1 Beta ◽  
Viral Control

Abstract Innate immunity triggers responsible for viral control or hyperinflammation in COVID- 19 are largely unknown. Here we show that the SARS-CoV-2 spike protein primes inflammasome activation and interleukin 1-beta (IL-1β) secretion in macrophages derived from COVID-19 patients but not in macrophages from healthy SARS-CoV-2 naïve controls. Chemical NLRP3 inhibition blocks spike protein-induced IL-1β secretion ex vivo. These findings can accelerate research on COVID-19 vaccine design and drug treatment.

scholarly journals P-MAPA, a Fungi-Derived Immunomodulatory Compound, Induces a Proinflammatory Response in a Human Whole Blood Model

2020 ◽  
Vol 2020 ◽  
pp. 1-13
Author(s):  
Mariana Torrente Gonçalves ◽  
Carla Cristina Squaiella-Baptistão ◽  
Giselle Pidde ◽  
Priscila Hess Lopes ◽  
Iseu da Silva Nunes ◽  
...  
Keyword(s):  
Animal Models ◽  
Complement Activation ◽  
Whole Blood ◽  
Cell Activation ◽  
Immune Cell ◽  
Ex Vivo ◽  
Proinflammatory Response ◽  
Human Whole Blood ◽  
Human Leukocytes ◽  
Immunomodulatory Properties

P-MAPA is a complex compound, derived from Aspergillus oryzae cultures, that has shown immunomodulatory properties in infection and cancer animal models. Despite promising results in these models, the mechanisms of cellular activation by P-MAPA, suggested to be Toll-like receptor- (TLR-) dependent, and its effect on human immune cells, remain unclear. Using an ex vivo model of human whole blood, the effects of P-MAPA on complement system activation, production of cytokines, and the expression of complement receptors (CD11b, C5aR, and C3aR), TLR2, TLR4, and the coreceptor CD14 were analyzed in neutrophils and monocytes. P-MAPA induced complement activation in human blood, detected by increased levels of C3a, C5a, and SC5b-9 in plasma. As a consequence, CD11b expression increased and C5aR decreased upon activation, while C3aR expression remained unchanged in leukocytes. TLR2 and TLR4 expressions were not modulated by P-MAPA treatment on neutrophils, but TLR4 expression was reduced in monocytes, while CD14 expression increased in both cell types. P-MAPA also induced the production of TNF-α, IL-8, and IL-12 and oxidative burst, measured by peroxynitrite levels, in human leukocytes. Complement inhibition with compstatin showed that P-MAPA-induced complement activation drives modulation of C5aR, but not of CD11b, suggesting that P-MAPA acts through both complement-dependent and complement-independent mechanisms. Compstatin also significantly reduced the peroxynitrite generation. Altogether, our results show that P-MAPA induced proinflammatory response in human leukocytes, which is partially mediated by complement activation. Our data contribute to elucidate the complement-dependent and complement-independent mechanisms of P-MAPA, which ultimately result in immune cell activation and in its immunomodulatory properties in infection and cancer animal models.

Elevated IL-1β contributes to antibody suppression produced by stress

2002 ◽  
Vol 93 (1) ◽  
pp. 207-215 ◽  
Author(s):  
Albert Moraska ◽  
Jay Campisi ◽  
Kien T. Nguyen ◽  
Steven F. Maier ◽  
Linda R. Watkins ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Acute Stress ◽  
Ex Vivo ◽  
Interleukin 1 ◽  
Keyhole Limpet Hemocyanin ◽  
Acquired Immunity ◽  
Sprague Dawley

Acute stressor exposure can facilitate innate immunity and suppress acquired immunity. The present study further characterized the potentiating effect of stress on innate immunity, interleukin-1β (IL-1β), and demonstrated that stress-induced potentiation of innate immunity may contribute to the stress-induced suppression of acquired immunity. The long-term effect of stress on IL-1β was measured by using an ex vivo approach. Sprague-Dawley rats were challenged with lipopolysaccharide (LPS) in vivo, and the IL-1β response was measured in vitro. Splenocytes, mesenteric lymphocytes, and peritoneal cavity cells had a dose- and time-dependent ex vivo IL-1β response to LPS. Rats that were exposed to inescapable shock (IS, 100 1.6 mA, 5-s tail shocks, 60-s intertrial interval) and challenged with a submaximal dose of LPS 4 days later had elevated IL-1β measured ex vivo. To test whether the acute stress-induced elevation in IL-1β contributes to the long-term suppression in acquired immunity, IL-1β receptors were blocked for 24 h after stress. Serum anti-keyhole limpet hemocyanin (KLH) immunoglobulin (Ig) was measured. In addition, the acute elevation (2 h post-IS) of splenic IL-1β in the absence of antigen was verified. Interleukin-1 receptor antagonist prevented IS-induced suppression in anti-KLH Ig. These data support the hypothesis that stress-induced increases in innate immunity (i.e., IL-1β) may contribute to stress-induced suppression in acquired immunity (i.e., anti-KLH Ig).

Explants of Intact Endometrium to Model Bovine Innate Immunity and Inflammation Ex Vivo

2012 ◽  
Vol 67 (6) ◽  
pp. 526-539 ◽  
Author(s):  
Álan Maia Borges ◽  
Gareth David Healey ◽  
Iain Martin Sheldon
Keyword(s):  
Innate Immunity ◽  
Ex Vivo

scholarly journals MicroRNA 21 (miR-21) and miR-181b Couple with NFI-A To Generate Myeloid-Derived Suppressor Cells and Promote Immunosuppression in Late Sepsis

2014 ◽  
Vol 82 (9) ◽  
pp. 3816-3825 ◽  
Author(s):  
Clara McClure ◽  
Laura Brudecki ◽  
Donald A. Ferguson ◽  
Zhi Q. Yao ◽  
Jonathan P. Moorman ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Innate Immunity ◽  
Transforming Growth Factor ◽  
Ex Vivo ◽  
Cecal Ligation ◽  
Myeloid Cells ◽  
Suppressor Cells ◽  
Myeloid Derived Suppressor Cells ◽  
Cell Repertoire ◽  
Myeloid Progenitors

ABSTRACTThe sepsis initial hyperinflammatory reaction, if not treated early, shifts to a protracted state of immunosuppression that alters both innate and adaptive immunity and is associated with elevated mortality. Myeloid-derived suppressor cells (MDSCs) are myeloid progenitors and precursors that fail to differentiate into mature innate-immunity cells and are known for their potent immunosuppressive activities. We previously reported that murine MDSCs expand dramatically in the bone marrow during late sepsis, induced by cecal ligation and puncture, and demonstrated that they contribute to late-sepsis immunosuppression. However, the molecular mechanism responsible for generating these immature Gr1+CD11b+myeloid cells during sepsis remains unknown. We show here that sepsis generates a microRNA (miRNA) signature that expands MDSCs. We found that miRNA 21 (miR-21) and miR-181b expression is upregulated in early sepsis and sustained in late sepsis. Importantly, we found that simultaneousin vivoblockade of both miRNAs via antagomiR (a chemically modified miRNA inhibitor) injection after sepsis initiation decreased the bone marrow Gr1+CD11b+myeloid progenitors, improved bacterial clearance, and reduced late-sepsis mortality by 74%. Gr1+CD11b+cells isolated from mice injected with antagomiRs were able to differentiateex vivointo macrophages and dendritic cells and produced smaller amounts of the immunosuppressive interleukin 10 (IL-10) and transforming growth factor β (TGF-β) after stimulation with bacterial lipopolysaccharide, suggesting that immature myeloid cells regained their maturation potential and have lost their immunosuppressive activity. In addition, we found that the protein level of transcription factor NFI-A, which plays a role in myeloid cell differentiation, was increased during sepsis and that antagomiR injection reduced its expression. Moreover, knockdown of NFI-A in the Gr1+CD11b+cells isolated from late-septic mice increased their maturation potential and reduced their production of the immunosuppressive mediators, similar to antagomiR injection. These data support the hypothesis that sepsis reprograms myeloid cells and thus alters the innate immunity cell repertoire to promote immunosuppression, and they demonstrate that this process can be reversed by targeting miR-21 and miR-181b to improve late-sepsis survival.

scholarly journals Prenatal vitamin D3supplementation suppresses LL-37 peptide expression inex vivoactivated neonatal macrophages but not their killing capacity

2014 ◽  
Vol 112 (6) ◽  
pp. 908-915 ◽  
Author(s):  
Rubhana Raqib ◽  
Anna Ly ◽  
Evana Akhtar ◽  
Akhirunnesa Mily ◽  
Nandita Perumal ◽  
...  
Keyword(s):  
Vitamin D ◽  
Innate Immunity ◽  
Cord Blood ◽  
Pregnant Women ◽  
Ex Vivo ◽  
Controlled Trial ◽  
Extracellular Fluid ◽  
Weekly Dose ◽  
Expression Ratio ◽  
Bactericidal Capacity

Vitamin D has regulatory effects on innate immunity. In the present study, we aimed to assess the effect of prenatal vitamin D3(vitD3) supplementation on neonatal innate immunity in a randomised, placebo-controlled trial by evaluating cathelicidin (LL-37) expression and the killing capacity of macrophages. Healthy pregnant women (n129) attending a clinic in Dhaka were randomised to receive either a weekly oral dose of 0·875 mg vitD3or placebo starting from 26 weeks of gestation up to delivery. Serum, plasma and monocyte-derived macrophages (MDM) were obtained from the cord blood. 25-Hydroxyvitamin D (25(OH)D) concentration was measured in serum. MDM were stimulated with or without Toll-like-receptor 4 ligand (TLR4L). Innate immune function was assessed by measuring LL-37 peptide levels in the culture supernatant of MDM by ELISA, LL-37 transcript levels by quantitative PCR, andex vivobactericidal capacity of MDM. vitD3supplementation did not increase LL-37 peptide levels in plasma or in the extracellular fluid of macrophages with or without TLR4L induction. However, stimulated intracellular LL-37 expression (ratio of stimulated:unstimulated MDM) was significantly reduced in the vitamin D groupv. placebo (P= 0·02). Multivariate-adjusted analyses showed that intracellular LL-37 peptide concentration from stimulated MDM was inversely associated with 25(OH)D concentration in serum (P= 0·03). TLR4L stimulation increased the bactericidal capacity of MDM compared with the unstimulated ones (P= 0·01); however, there was no difference in killing capacity between the two groups. A weekly dose of 0·875 mg vitD3to healthy pregnant women suppressed the intracellular LL-37 peptide stores of activated macrophages, but did not significantly affect theex vivobactericidal capacity of cord blood MDM.

The effect of Intravenous Immunoglobulin (IVIG) on \textit{ex vivo} activation of human leukocytes

2017 ◽  
Vol 24 (3-4) ◽  
pp. 39-44 ◽  
Author(s):  
Liliya Yu Basyreva ◽  
Ilya B. Brodsky ◽  
Alexander A. Gusev ◽  
Olga N. Zhapparova ◽  
Elena V. Mikhalchik ◽  
...  
Keyword(s):  
Intravenous Immunoglobulin ◽  
Ex Vivo ◽  
Human Leukocytes

scholarly journals Platelet Toll-like Receptor 4-related Innate Immunity Potentially Participates in Transfusion Reactions Independent of ABO Compatibility: An ex Vivo Study

2021 ◽  
Author(s):  
Chien-Sung Tsai ◽  
Mei-Hua Hu ◽  
Yung-Chi Hsu ◽  
Go-Shine Huang
Keyword(s):  
Innate Immunity ◽  
Ex Vivo ◽  
Binding Protein ◽  
Blood Type ◽  
Saline Control ◽  
Dependent Manner ◽  
Tlr4 Expression ◽  
Tnf Α ◽  
Transfusion Reactions ◽  
Lps Binding

Abstract Purpose: The role of platelet TLR4 in transfusion reactions remains unclear. This study analyzed platelet TLR4, certain DAMPs, and the effect of ABO compatibility on TLR4 expression after a simulated transfusion ex vivo.Methods: Donor red blood cells were harvested from a blood bank. Recipient blood from patients undergoing cardiac surgery was processed to generate a washed platelet suspension. Donor blood was added to the washed platelets at 1%, 5%, or 10% (v/v). Blood mixing experiments were performed using four groups: 0.9% saline control group (n = 31); M, matched blood type mixing (n = 20); S, uncross-matched ABO type-specific mixing (n = 20); and I, ABO incompatible blood mixing (n = 20). Platelet TLR4 expression was determined after blood mixing. Levels of TLR4-binding DAMPs (HMGB1, S100A8, S100A9, and SAA) and that of LPS-binding protein and endpoint proteins (TNF-α, IL-1β, and IL-6) in the TLR4 signaling pathway were evaluated.Results: The 1%, 5%, and 10% blood mixtures significantly increased TLR4 expression in three groups (M, S, and I; all P < 0.001) in a concentration-dependent manner. TLR4 expression did not significantly differ among the three groups (P = 0.148). HMGB1, S100A8, and S100A9 showed elevated levels in response to blood mixing; SAA, LPS-binding protein, TNF-α, IL-1β, and IL-6 did not. Conclusion: Blood mixing may elicit innate immune responses by upregulating platelet TLR4 and DAMPs unassociated with ABO compatibility, suggesting that innate immunity through TLR4-mediated signaling may induce transfusion reactions. The trial was retrospectively registered at Chinese Clinical Trial Registry (ChiCTR2100045606) with date of registration on 19 April 2021.

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