The cathelicidin CATH-2 efficiently neutralizes LPS- and E. coli-induced activation of porcine bone marrow derived macrophages

The roots and rhizomes ofGlycyrrhizaspecies (licorice) have been widely used as natural sweeteners and herbal medicines. The aim of this study is to investigate the effect of glycyrrhizic acid (GA) from licorice on macrophage polarization. Both phenotypic and functional activities of murine bone marrow-derived macrophages (BMDMs) treated by GA were assessed. Our results showed that GA obviously increased the cell surface expression of CD80, CD86, and MHCII molecules. Meanwhile, GA upregulated the expression of CCR7 and the production of TNF-α, IL-12, IL-6, and NO (the markers of classically activated (M1) macrophages), whereas it downregulated the expression of MR, Ym1, and Arg1 (the markers of alternatively activated (M2) macrophage). The functional tests showed that GA dramatically enhanced the uptake of FITC-dextran andE. coliK88 by BMDMs and decreased the intracellular survival ofE. coliK88 andS. typhimurium. Moreover, we demonstrated that JNK and NF-κB activation are required for GA-induced NO and M1-related cytokines production, while ERK1/2 pathway exhibits a regulatory effect via induction of IL-10. Together, these findings indicated that GA promoted polarization of M1 macrophages and enhanced its phagocytosis and bactericidal capacity. The results expanded our knowledge about the role of GA in macrophage polarization.

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TLR Activation Alters Bone Marrow-Derived Macrophage Differentiation

Journal of Innate Immunity ◽

10.1159/000494070 ◽

2018 ◽

Vol 11 (1) ◽

pp. 99-108 ◽

Cited By ~ 1

Author(s):

Gertrude O. Oppong-Nonterah ◽

Omar Lakhdari ◽

Asami Yamamura ◽

Hal M. Hoffman ◽

Lawrence S. Prince

Keyword(s):

Bone Marrow ◽

Early Stage ◽

Macrophage Differentiation ◽

E Coli ◽

Inflammatory Signals ◽

Macrophage Marker ◽

Macrophage Phagocytosis ◽

And Function ◽

Induced Changes ◽

Lps Treatment

Early exposure to inflammatory signals may have a lasting impact on immune function. Present throughout embryogenesis, macrophages are key cells providing innate immune protection to the developing fetus and newborn. Here, we have used an established model of macrophage development to test how early inflammatory signals can impact cellular differentiation and function. Bone marrow-derived macrophages were treated with Escherichia coli lipopolysaccharide (LPS) 2 days after initial isolation and culture. LPS treatment during this early stage of differentiation decreased the expression of CSF1R and increased that of the mature macrophage marker F4/80. These early changes in macrophage differentiation were also measured in cells from mice lacking IKKβ, but the change in CSF1R expression after LPS treatment was blocked with MAPK inhibition. LPS-induced changes in macrophage marker expression persisted following LPS removal, suggesting that early inflammatory activation could induce a lasting developmental impact. Early LPS exposure inhibited macrophage phagocytosis of labeled E. coli while LPS had no effect on fully differentiated macrophages. Our data demonstrate that early inflammatory exposure to a microbial stimulus induce lasting phenotypic changes in macrophages.

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Differential immunomodulation of porcine bone marrow derived dendritic cells by E. coli Nissle 1917 and β-glucans

PLoS ONE ◽

10.1371/journal.pone.0233773 ◽

2020 ◽

Vol 15 (6) ◽

pp. e0233773

Author(s):

Mirelle Geervliet ◽

Laura C. P. Lute ◽

Christine A. Jansen ◽

Victor P. M. G. Rutten ◽

Huub F. J. Savelkoul ◽

...

Keyword(s):

Bone Marrow ◽

Dendritic Cells ◽

E Coli

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‘Cleaning’ autologous bone marrow transplants with E. coli toxin

Drug Discovery Today ◽

10.1016/s1359-6446(99)01467-1 ◽

2000 ◽

Vol 5 (3) ◽

pp. 91-92

Author(s):

Janet Fricker

Keyword(s):

Bone Marrow ◽

Autologous Bone Marrow ◽

Bone Marrow Transplants ◽

E Coli ◽

Autologous Bone

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Immunocytochemical characteristics and ultrastructures of immature B cells in bone marrow of rats administered IV with E. Coli and S. Typhimurium lipopolysaccharides (LPS)

International Journal of Immunopharmacology ◽

10.1016/0192-0561(88)90441-9 ◽

1988 ◽

Vol 10 ◽

pp. 113

Author(s):

H. Kawaji ◽

H. Maeda ◽

T. Matsuura ◽

H. Nakajima ◽

W. Kurio ◽

...

Keyword(s):

Bone Marrow ◽

B Cells ◽

E Coli ◽

Immature B Cells

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Toll-Like Receptor 4/Stem Cell Antigen 1 Signaling Promotes Hematopoietic Precursor Cell Commitment to Granulocyte Development during the Granulopoietic Response to Escherichia coli Bacteremia

Infection and Immunity ◽

10.1128/iai.01280-12 ◽

2013 ◽

Vol 81 (6) ◽

pp. 2197-2205 ◽

Cited By ~ 17

Author(s):

Xin Shi ◽

Robert W. Siggins ◽

William L. Stanford ◽

John N. Melvan ◽

Marc D. Basson ◽

...

Keyword(s):

Escherichia Coli ◽

Bone Marrow ◽

Stem Cell ◽

Precursor Cell ◽

Toll Like Receptor ◽

Toll Like Receptor 4 ◽

Content Type ◽

E Coli ◽

Hematopoietic Precursor

ABSTRACTIn response to severe bacterial infection, bone marrow hematopoietic activity shifts toward promoting granulopoiesis. The underlying cell signaling mechanisms remain obscure. To study the role of Toll-like receptor 4 (TLR4)/stem cell antigen-1 (Sca-1) signaling in this process, bacteremia was induced in mice by intravenous injection ofEscherichia coli. A subgroup of animals also received intravenous 5-bromo-2-deoxyuridine (BrdU). In a separate set of experiments, bone marrow lineage-negative (lin−) stem cell growth factor receptor-positive (c-kit+) Sca-1−cells containing primarily common myeloid progenitors were culturedin vitrowithout or withE. colilipopolysaccharide (LPS). In genotypic background control mice, bacteremia significantly upregulated Sca-1 expression by lin−c-kit+cells, as reflected by a marked increase in BrdU-negative lin−c-kit+Sca-1+cells in the bone marrow. In mice with the TLR4 gene deletion, this bacteremia-evoked Sca-1 response was blocked.In vitro, LPS induced a dose-dependent increase in Sca-1 expression by cultured marrow lin−c-kit+Sca-1−cells. LPS-induced upregulation of Sca-1 expression was regulated at the transcriptional level. Inhibition of c-Jun N-terminal kinase/stress-activated protein kinase (JNK) activity with the specific inhibitor SP600125 suppressed LPS-induced upregulation of Sca-1 expression by marrow lin−c-kit+Sca-1−cells. Engagement of Sca-1 with anti-Sca-1 antibodies enhanced the expression of Sfpi1 spleen focus-forming virus (SFFV) proviral integration 1 (PU.1) in marrow lin−c-kit+Sca-1−cells cultured with LPS. Sca-1 null mice failed to maintain the marrow pool of granulopoietic cells following bacteremia. These results demonstrate that TLR4/Sca-1 signaling plays an important role in the regulation of hematopoietic precursor cell programming and their enhancement of granulocyte lineage commitment in response toE. colibacteremia.

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Optimizing Granulocyte Colony-Stimulating Factor Transcript for Enhanced Expression in Escherichia coli

Frontiers in Bioengineering and Biotechnology ◽

10.3389/fbioe.2021.630367 ◽

2021 ◽

Vol 9 ◽

Author(s):

Sonal Datta

Keyword(s):

Escherichia Coli ◽

Bone Marrow ◽

Protein Expression ◽

Therapeutic Proteins ◽

Secondary Structures ◽

Granulocyte Colony Stimulating Factor ◽

Colony Stimulating Factor ◽

E Coli ◽

Enhanced Expression ◽

Stimulating Factor

The human granulocyte colony-stimulating factor (G-CSF) is a hematopoietic growth factor used to prevent and treat neutropenia. G-CSF stimulates the bone marrow to produce infection-fighting granulocytes. Food and Drug Administration of the United States approved G-CSF in 1991 and its PEGylated version in 2002 as a prophylactic and therapeutic measure against neutropenia. Recombinant human G-CSF is produced in surrogate host Escherichia coli and is PEGylated at N-terminal. Besides neutropenia, G-CSF is also used in bone marrow transplantation for the mobilization and maturation of peripheral blood stem cells. Considering the requirement of producing G-CSF therapeutic in large quantities, construct designing for high expression is critical for the biopharmaceutical and industrial application. Earlier studies have employed approaches such as codon optimization, use of strong promoters, employment of protein tags, secretion signals, optimization of protein folding, etc., for increasing expression and yield of therapeutic proteins. In this study, it was observed that mRNA transcribed from the native human cDNA of G-CSF and the codon-optimized variant leads to low protein expression in E. coli. To understand the underlying reasons, the mRNA secondary structure of the 5′ end of the G-CSF transcript was analyzed. This analysis revealed the presence of stable secondary structures at the 5′ end of the G-CSF transcript, arising from the native human gene and even from the codon-optimized sequence. These secondary structures were disrupted through translationally silent mutations within the first 24 nucleotides of the transcript without affecting the protein sequence. Interestingly, through this approach, the G-CSF protein expression was increased 60 folds as compared to native G-CSF construct. We believe that these findings create a roadmap for optimization of G-CSF transcript for enhanced expression in E. coli and could be employed to increase the expression of other therapeutic proteins.

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Group V Phospholipase A2 in Bone Marrow-derived Myeloid Cells and Bronchial Epithelial Cells Promotes Bacterial Clearance after Escherichia coli Pneumonia

Journal of Biological Chemistry ◽

10.1074/jbc.m111.262733 ◽

2011 ◽

Vol 286 (41) ◽

pp. 35650-35662 ◽

Cited By ~ 13

Author(s):

Norbert Degousee ◽

David J. Kelvin ◽

Gerd Geisslinger ◽

David M. Hwang ◽

Eva Stefanski ◽

...

Keyword(s):

Escherichia Coli ◽

Bone Marrow ◽

Epithelial Cells ◽

Bronchoalveolar Lavage ◽

Myeloid Cells ◽

Respiratory Acidosis ◽

Bronchial Epithelial Cells ◽

Group V ◽

Bronchial Epithelial ◽

E Coli

Group V-secreted phospholipase A2 (GV sPLA2) hydrolyzes bacterial phospholipids and initiates eicosanoid biosynthesis. Here, we elucidate the role of GV sPLA2 in the pathophysiology of Escherichia coli pneumonia. Inflammatory cells and bronchial epithelial cells both express GV sPLA2 after pulmonary E. coli infection. GV−/− mice accumulate fewer polymorphonuclear leukocytes in alveoli, have higher levels of E. coli in bronchoalveolar lavage fluid and lung, and develop respiratory acidosis, more severe hypothermia, and higher IL-6, IL-10, and TNF-α levels than GV+/+ mice after pulmonary E. coli infection. Eicosanoid levels in bronchoalveolar lavage are similar in GV+/+ and GV−/− mice after lung E. coli infection. In contrast, GV+/+ mice have higher levels of prostaglandin D2 (PGD2), PGF2α, and 15-keto-PGE2 in lung and express higher levels of ICAM-1 and PECAM-1 on pulmonary endothelial cells than GV−/− mice after lung infection with E. coli. Selective deletion of GV sPLA2 in non-myeloid cells impairs leukocyte accumulation after pulmonary E. coli infection, and lack of GV sPLA2 in either bone marrow-derived myeloid cells or non-myeloid cells attenuates E. coli clearance from the alveolar space and the lung parenchyma. These observations show that GV sPLA2 in bone marrow-derived myeloid cells as well as non-myeloid cells, which are likely bronchial epithelial cells, participate in the regulation of the innate immune response to pulmonary infection with E. coli.

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