scholarly journals A primary human macrophage-enteroid co-culture model to investigate mucosal gut physiology and host-pathogen interactions

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Gaelle Noel ◽  
Nicholas W. Baetz ◽  
Janet F. Staab ◽  
Mark Donowitz ◽  
Olga Kovbasnjuk ◽  
...  
Keyword(s):  
Intestinal Epithelium ◽  
Immune Cells ◽  
Barrier Function ◽  
Morphological Changes ◽  
Human Monocyte ◽  
Human Macrophage ◽  
Primary Human Macrophage ◽  
Cell Height ◽  
Culture Model ◽  
Gut Physiology

Abstract Integration of the intestinal epithelium and the mucosal immune system is critical for gut homeostasis. The intestinal epithelium is a functional barrier that secludes luminal content, senses changes in the gut microenvironment, and releases immune regulators that signal underlying immune cells. However, interactions between epithelial and innate immune cells to maintain barrier integrity and prevent infection are complex and poorly understood. We developed and characterized a primary human macrophage-enteroid co-culture model for in-depth studies of epithelial and macrophage interactions. Human intestinal stem cell-derived enteroid monolayers co-cultured with human monocyte-derived macrophages were used to evaluate barrier function, cytokine secretion, and protein expression under basal conditions and following bacterial infection. Macrophages enhanced barrier function and maturity of enteroid monolayers as indicated by increased transepithelial electrical resistance and cell height. Communication between the epithelium and macrophages was demonstrated through morphological changes and cytokine production. Intraepithelial macrophage projections, efficient phagocytosis, and stabilized enteroid barrier function revealed a coordinated response to enterotoxigenic and enteropathogenic E. coli infections. In summary, we have established the first primary human macrophage-enteroid co-culture system, defined conditions that allow for a practical and reproducible culture model, and demonstrated its suitability to study gut physiology and host responses to enteric pathogens.

A Primary Human Macrophage-Enteroid Co-Culture Model to Investigate Mucosal Gut Physiology and Host-Pathogen Interactions

2017 ◽  
Vol 152 (5) ◽  
pp. S56-S57
Author(s):  
Nicholas W. Baetz ◽  
Gaelle Noel ◽  
Janet Staab ◽  
Mark Donowitz ◽  
Olga Kovbasnjuk ◽  
...  
Keyword(s):  
Human Macrophage ◽  
Primary Human Macrophage ◽  
Host Pathogen Interactions ◽  
Culture Model ◽  
Host Pathogen ◽  
Gut Physiology

scholarly journals Erratum: A primary human macrophage-enteroid co-culture model to investigate mucosal gut physiology and host-pathogen interactions

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Gaelle Noel ◽  
Nicholas W. Baetz ◽  
Janet F. Staab ◽  
Mark Donowitz ◽  
Olga Kovbasnjuk ◽  
...  
Keyword(s):  
Human Macrophage ◽  
Primary Human Macrophage ◽  
Host Pathogen Interactions ◽  
Culture Model ◽  
Host Pathogen ◽  
Gut Physiology

scholarly journals Translation of Collagen Ultrastructure to Biomaterial Fabrication for Material Independent but Highly Efficient Topographic Immunomodulation

2021 ◽  
Author(s):  
matthias ryma ◽  
tina tylek ◽  
julia liebscher ◽  
robin fernandez ◽  
christoph böhm ◽  
...  
Keyword(s):  
Immune Cells ◽  
Macrophage Polarization ◽  
Healing Process ◽  
Human Monocyte ◽  
Collagen Fibers ◽  
Collagen I ◽  
Human Macrophage ◽  
Type I ◽  
Native Collagen ◽  
Surface Patterns

<div>Supplement-free induction of cellular differentiation and polarization solely through the topography of materials is an auspicious strategy but has so far significantly lacked behind the efficiency and intensity of media-supplementation based protocols. For immune cells, low intensity effects were achieved on rhodent cells using standard technologically driven surface patterns and scaffold geometries, but no effects could be achieved for human immune cells. <br></div><p> </p> <p>Consistent with the idea that 3D structural motives in the extracellular matrix possess immunomodulatory capacity as part of the natural healing process, we found that human monocyte-derived macrophages show a strong M2a like pro-healing polarization when cultured on type I rat-tail collagen fibers (hereafter termed "collagen I") but not on collagen I films. <br></p><p>Therefore, we hypothesized that highly aligned nanofibrils also of synthetic polymers, if packed into larger bundles in 3D topographical similarity to native collagen I, would induce a localized macrophage polarization. <br></p> <p> </p> <p>For the automated fabrication of such bundles in a 3D printing manner, we pioneered the strategy of "Melt Electrofibrillation" by the integration of flow directed polymer phase separation into Melt Electrowriting and subsequent selective dissolution of the matrix polymer. This process yields nano-fiber bundles with a remarkable structural similarity to native collagen I fibers, particularly for medical grade polycaprolactone (PCL). </p> <p> </p> <p>These biomimetic fibrillar structures indeed induced a pronounced elongation of human monocyte-derived macrophages and unprecedentedly triggered their M2-like polarization as efficiently as IL-4 cytokine treatment.</p> <p> </p> <p>Our data evidence the biological importance of human macrophage-elongation on collagen fibers and pioneers a strategy to fabricate scaffolds that exploit this effect to drive macrophage polarization through precise and biomimetic material design. </p>

scholarly journals The intestinal immunoendocrine axis: novel cross-talk between enteroendocrine cells and the immune system during infection and inflammatory disease

2015 ◽  
Vol 43 (4) ◽  
pp. 727-733 ◽  
Author(s):  
John J Worthington
Keyword(s):  
Immune System ◽  
Cross Talk ◽  
Intestinal Epithelium ◽  
Immune Cells ◽  
Inflammatory Disease ◽  
Barrier Function ◽  
Crucial Role ◽  
Enteroendocrine Cells ◽  
Immune Homeostasis

The intestinal epithelium plays a crucial role in maintaining barrier function and immune homeostasis, a failure of which results in disease. This review focuses on the epithelial enteroendocrine cells and the crosstalk that exists with immune cells during inflammation.

scholarly journals Translation of Collagen Ultrastructure to Biomaterial Fabrication for Material Independent but Highly Efficient Topographic Immunomodulation

2021 ◽  
Author(s):  
matthias ryma ◽  
tina tylek ◽  
julia liebscher ◽  
robin fernandez ◽  
christoph böhm ◽  
...  
Keyword(s):  
Immune Cells ◽  
Macrophage Polarization ◽  
Healing Process ◽  
Human Monocyte ◽  
Collagen Fibers ◽  
Collagen I ◽  
Human Macrophage ◽  
Type I ◽  
Native Collagen ◽  
Surface Patterns

<div>Supplement-free induction of cellular differentiation and polarization solely through the topography of materials is an auspicious strategy but has so far significantly lacked behind the efficiency and intensity of media-supplementation based protocols. For immune cells, low intensity effects were achieved on rhodent cells using standard technologically driven surface patterns and scaffold geometries, but no effects could be achieved for human immune cells. <br></div><p> </p> <p>Consistent with the idea that 3D structural motives in the extracellular matrix possess immunomodulatory capacity as part of the natural healing process, we found that human monocyte-derived macrophages show a strong M2a like pro-healing polarization when cultured on type I rat-tail collagen fibers (hereafter termed "collagen I") but not on collagen I films. <br></p><p>Therefore, we hypothesized that highly aligned nanofibrils also of synthetic polymers, if packed into larger bundles in 3D topographical similarity to native collagen I, would induce a localized macrophage polarization. <br></p> <p> </p> <p>For the automated fabrication of such bundles in a 3D printing manner, we pioneered the strategy of "Melt Electrofibrillation" by the integration of flow directed polymer phase separation into Melt Electrowriting and subsequent selective dissolution of the matrix polymer. This process yields nano-fiber bundles with a remarkable structural similarity to native collagen I fibers, particularly for medical grade polycaprolactone (PCL). </p> <p> </p> <p>These biomimetic fibrillar structures indeed induced a pronounced elongation of human monocyte-derived macrophages and unprecedentedly triggered their M2-like polarization as efficiently as IL-4 cytokine treatment.</p> <p> </p> <p>Our data evidence the biological importance of human macrophage-elongation on collagen fibers and pioneers a strategy to fabricate scaffolds that exploit this effect to drive macrophage polarization through precise and biomimetic material design. </p>

Abstract 516: Knockdown of the Hypertension Associated Gene NOSTRIN Alters Glomerular Barrier Function in Zebrafish (Danio rerio)

Hypertension ◽  
2012 ◽  
Vol 60 (suppl_1) ◽  
Author(s):  
Torsten Kirsch ◽  
Jessica Kaufeld ◽  
Ron Korstanje ◽  
Dirk Hentschel ◽  
Hermann Haller ◽  
...  
Keyword(s):  
Glomerular Filtration ◽  
Barrier Function ◽  
Morphological Changes ◽  
Glomerular Filtration Barrier ◽  
Cell Dysfunction ◽  
Larval Zebrafish ◽  
Filtration Barrier ◽  
Important Regulator ◽  
Glomerular Barrier ◽  
Prime Source

The bioavailability of nitric oxide (NO) has been associated with the development and progression of vascular and renal disease. NOSTRIN (for eNOS Traffic Inducer) has primarily been recognized as one important regulator of eNOS, the prime source of NO in the cardiovascular system, with a possible role in the pathogenesis of pre-eclampsia and the development of increased intrahepatic resistance in liver disease. Here, we identified NOSTRIN in the center of a QTL-overlap region in rat and human trait loci that are associated with hypertension. Glomerular NOSTRIN expression is detectable in podocytes in human and rat glomeruli and podocytic NOSTRIN expression is diminished in hypertensive kidney disease. We show that knockdown of NOSTRIN alters the glomerular filtration barrier function in larval zebrafish, inducing proteinuria and leading to ultrastructural morphological changes on the endothelial as well as epithelial side and the GBM of the glomerular capillary loop. We also demonstrate that NOSTRIN interacts with proteins associated with the podocyte slit membrane. We conclude that NOSTRIN expression is an important factor for the integrity of the glomerular filtration barrier. Disease related alteration of NOSTRIN expression may not only affect the vascular endothelium and therefore contribute to endothelial cell dysfunction but may also contribute to the development of podocyte disease and proteinuria.

scholarly journals A Novel Model of Cancer-Induced Peripheral Neuropathy and the Role of TRPA1 in Pain Transduction

2017 ◽  
Vol 2017 ◽  
pp. 1-12 ◽  
Author(s):  
Ahmad Maqboul ◽  
Bakheet Elsadek
Keyword(s):  
Immune Cells ◽  
Peripheral Nerves ◽  
Morphological Changes ◽  
Nerve Fibres ◽  
Cold Allodynia ◽  
Sciatic Nerves ◽  
Pain Transduction ◽  
Thermal Stimuli ◽  
Novel Model

Background. Models of cancer-induced neuropathy are designed by injecting cancer cells near the peripheral nerves. The interference of tissue-resident immune cells does not allow a direct contact with nerve fibres which affects the tumor microenvironment and the invasion process. Methods. Anaplastic tumor-1 (AT-1) cells were inoculated within the sciatic nerves (SNs) of male Copenhagen rats. Lumbar dorsal root ganglia (DRGs) and the SNs were collected on days 3, 7, 14, and 21. SN tissues were examined for morphological changes and DRG tissues for immunofluorescence, electrophoretic tendency, and mRNA quantification. Hypersensitivities to cold, mechanical, and thermal stimuli were determined. HC-030031, a selective TRPA1 antagonist, was used to treat cold allodynia. Results. Nociception thresholds were identified on day 6. Immunofluorescent micrographs showed overexpression of TRPA1 on days 7 and 14 and of CGRP on day 14 until day 21. Both TRPA1 and CGRP were coexpressed on the same cells. Immunoblots exhibited an increase in TRPA1 expression on day 14. TRPA1 mRNA underwent an increase on day 7 (normalized to 18S). Injection of HC-030031 transiently reversed the cold allodynia. Conclusion. A novel and a promising model of cancer-induced neuropathy was established, and the role of TRPA1 and CGRP in pain transduction was examined.

Abstract 375: The Intergenic Long Non-Coding RNA RP11-472n13.3 Modulates Interferon-Gamma Signaling and M1 Macrophage Activation

2017 ◽  
Vol 37 (suppl_1) ◽  
Author(s):  
Ying Wang ◽  
Chenyi Xue ◽  
Muredach Reilly ◽  
Hanrui Zhang
Keyword(s):  
Molecular Mechanisms ◽  
Target Genes ◽  
Induced Pluripotent Stem Cell ◽  
Myeloid Cell ◽  
Human Monocyte ◽  
Human Macrophage ◽  
Nucleotide Polymorphisms ◽  
Non Coding Rna ◽  
M1 Macrophage ◽  
Long Non Coding Rna

We aim to interrogate the functions of a subset of human macrophage intergenic long non-coding RNA (lincRNAs) which harbor cardiometabolic trait-associated single nucleotide polymorphisms (SNPs). We have found that one lincRNA RP11-472N13.3 overlaps rs7081678, a SNP significantly associated with central obesity (WHRadjBMI; P =5.57x10 -6 ). RP11-472N13.3 expression is enriched in macrophages relative to other obesity relevant tissues. Thus, RP11-472N13.3 SNPs for obesity may act via its myeloid cell modulation in adipose. In human monocyte-derived macrophage (HMDM), human induced pluripotent stem cell-derived macrophages (IPSDM) and THP1-derived macrophages (THP-1Φ), at RNAseq and Q-PCR, RP11-472N13.3 is abundant in M0 and M2(IL-4) macrophages but markedly suppressed in the M1 state (LPS/IFNγ). RP11-472N13.3 localizes almost exclusively to the cytoplasmic fraction of M0-HMDM. Consistent with GENCODE, our HMDM RNAseq data suggest a single 2-exon isoform. ChIP-seq reveals PU.1 and C/EBP-β binding at RP11-472N13.3 transcription start site. In our HMDM RNAseq (n=30 subjects) data, RP11-472N13.3 expression was inversely correlated with IFNγ-JAK-STAT signaling genes (e.g., IRF4, IL-12A, IL-23, STAT1, SOCS1, SOCS3 ), but not LPS/TLR4 activated genes (e.g., TNFA, CXCL9, CXCL10, IL1B ). Furthermore, KD of RP11-472N13.3 using siRNA or LNA-ASO in THP-1Φ, amplified expression of IFNγ target genes but not LPS/TLR4 targets during M1 activation (LPS/IFNγ). These data suggest its potential role in modulating IFNγ signaling. Mechanistic studies are needed to examine the molecular mechanisms.

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