Chemotactic and Immunoregulatory Properties of Bone Cells are Modulated by Endotoxin-Stimulated Lymphocytes

Cvija, Hrvoje; Kovačić, Nataša; Katavić, Vedran; Ivčević, Sanja; Aguila, Héctor L.; Marušić, Ana; Grčević, Danka

doi:10.1007/s10753-012-9477-y

Cited by 4 publications

(8 citation statements)

References 47 publications

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“…Spleen and peripheral blood mononuclear cells [PBMC; obtained by Histopaque (Sigma‐Aldrich) separation] were cultured in 48‐well plates at a density of 0·5 × 10 6 /well in 0·5 ml/well of α‐MEM/10% FCS supplemented with 20 ng/ml M‐CSF and 40 ng/mL RANKL. At days 5–7 of culture, tartrate‐resistant acid phosphatase (TRAP)‐positive multi‐nucleated osteoclasts (≥ three nuclei/cell) were identified using a commercially available kit (Sigma‐Aldrich) and counted by light microscopy . In selected experiments, osteoclast cultures were treated with 10 ng/ml CCL5 (PeproTech, Rocky Hill, NJ, USA).…”

Section: Methodsmentioning

confidence: 99%

“…Sorting purity was determined by a reanalysis of fractioned populations and was greater than 99% for all experiments. At days 5–7 of culture, TRAP + multi‐nucleated osteoclasts (≥ three nuclei/cell) were counted by light microscopy .…”

Section: Methodsmentioning

confidence: 99%

“…Expression of osteoclast differentiation genes [cFms/colony stimulating factor 1 receptor ( Csf1r ), Rank , cathepsin K ( Ctsk ), calcitonin receptor ( Calcr )], chemokines ( Ccl2 , Ccl5 ) and cytokines ( Il6 , Tnf ) was determined using commercially available TaqMan Gene Expression Assays (Applied Biosystems). The expression of a specific gene was calculated according to the relative standard curve of gene expression in the calibrator sample (cDNA from mononuclear cells or osteoclast culture), and then normalized to the expression level of the β‐actin gene as an endogenous control .…”

Section: Methodsmentioning

confidence: 99%

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Increased chemotaxis and activity of circulatory myeloid progenitor cells may contribute to enhanced osteoclastogenesis and bone loss in the C57BL/6 mouse model of collagen-induced arthritis

Matijašević

Flegar

Kovačić

et al. 2016

Clinical and Experimental Immunology

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Our study aimed to determine the functional activity of different osteoclast progenitor (OCP) subpopulations and signals important for their migration to bone lesions, causing local and systemic bone resorption during the course of collagen-induced arthritis in C57BL/6 mice. Arthritis was induced with chicken type II collagen (CII), and assessed by clinical scoring and detection of anti-CII antibodies. We observed decreased trabecular bone volume of axial and appendicular skeleton by histomorphometry and micro-computed tomography as well as decreased bone formation and increased bone resorption rate in arthritic mice in vivo. In the affected joints, bone loss was accompanied with severe osteitis and bone marrow hypercellularity, coinciding with the areas of active osteoclasts and bone erosions. Flow cytometry analysis showed increased frequency of putative OCP cells (CD3 B220 NK1.1 CD11b CD117 CD115 for bone marrow and CD3 B220 NK1.1 CD11b CD115 Gr-1 for peripheral haematopoietic tissues), which exhibited enhanced differentiation potential in vitro. Moreover, the total CD11b population was expanded in arthritic mice as well as CD11b F4/80 macrophage, CD11b NK1.1 natural killer cell and CD11b CD11c myeloid dendritic cell populations in both bone marrow and peripheral blood. In addition, arthritic mice had increased expression of tumour necrosis factor-α, interleukin-6, CC chemokine ligand-2 (Ccl2) and Ccl5, with increased migration and differentiation of circulatory OCPs in response to CCL2 and, particularly, CCL5 signals. Our study characterized the frequency and functional properties of OCPs under inflammatory conditions associated with arthritis, which may help to clarify crucial molecular signals provided by immune cells to mediate systemically enhanced osteoresorption.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Increased chemotaxis and activity of circulatory myeloid progenitor cells may contribute to enhanced osteoclastogenesis and bone loss in the C57BL/6 mouse model of collagen-induced arthritis

Matijašević

Flegar

Kovačić

et al. 2016

Clinical and Experimental Immunology

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show abstract

“…Total RNA was extracted from isolated BM, spleen and homogenized bones (tibia) using TRI reagent (Applied Biosystems, Foster City, CA, USA) and reversely transcribed (1µg) to cDNA and amplified (20 ng/well) by quantitative (q)PCR using an ABI Prism 7500 Sequence detection system (Applied Biosystems) in a 25µL reaction volume [46]. Commercially available TaqMan Gene Expression Assays for osteoclast- analysis suggest that the least difference in gene expression that could be reproducibly detected is around 100% [46,47] and therefore we assume that a difference of 100% or more in gene expression, repeating through all experiments, is biologically significant.…”

Section: Gene Expression Analysismentioning

confidence: 99%

“…Commercially available TaqMan Gene Expression Assays for osteoclast- analysis suggest that the least difference in gene expression that could be reproducibly detected is around 100% [46,47] and therefore we assume that a difference of 100% or more in gene expression, repeating through all experiments, is biologically significant.…”

Section: Gene Expression Analysismentioning

confidence: 99%

Acute hematopoietic stress in mice is followed by enhanced osteoclast maturation in the bone marrow microenvironment

Kuzmac

Grčević

Šućur

et al. 2014

Experimental Hematology

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Recent advances in endotoxin tolerance

Liú

Cao

Zhou

et al. 2018

J of Cellular Biochemistry

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Endotoxin tolerance is defined as a reduced capacity of a cell to respond endotoxin (lipopolysaccharide, LPS) challenge after an initial encounter with endotoxin in advance. The body becomes tolerant to subsequent challenge with a lethal dose of endotoxin and cytokines release and cell/tissue damage induced by inflammatory reaction are significantly reduced in the state of endotoxin tolerance. The main characteristics of endotoxin tolerance are downregulation of inflammatory mediators such as tumor necrosis factor α (TNF‐α), interleukin‐1β (IL‐1β), and C‐X‐C motif chemokine 10 (CXCL10) and upregulation of anti‐inflammatory cytokines such as IL‐10 and transforming growth factor β (TGF‐β). Therefore, endotoxin tolerance is often regarded as the regulatory mechanism of the host against excessive inflammation. Endotoxin tolerance is a complex pathophysiological process and involved in multiple cellular signal pathways, receptor alterations, and biological molecules. However, the exact mechanism remains elusive up to date. To better understand the underlying cellular and molecular mechanisms of endotoxin tolerance, it is crucial to investigate the comprehensive cellular signal pathways, signaling proteins, cell surface molecules, proinflammatory and anti‐inflammatory cytokines, and other mediators. Endotoxin tolerance plays an important role in reducing the mortality of sepsis, endotoxin shock, and other endotoxin‐related diseases. Recent reports indicated that endotoxin tolerance is also related to other diseases such as cystic fibrosis, acute coronary syndrome, liver ischemia‐reperfusion injury, and cancer. The aim of this review is to discuss the recent advances in endotoxin tolerance mainly based on the cellular and molecular mechanisms by outline the current state of the knowledge of the involvement of the toll‐like receptor 4 (TLR4) signaling pathways, negative regulate factor, microRNAs, apoptosis, chromatin modification, and gene reprogramming of immune cells in endotoxin tolerance. We hope to provide a new idea and scientific basis for the rational treatment of endotoxin‐related diseases such as endotoxemia, sepsis, and endotoxin shock clinically.

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Chemotactic and Immunoregulatory Properties of Bone Cells are Modulated by Endotoxin-Stimulated Lymphocytes

Cited by 4 publications

References 47 publications

Increased chemotaxis and activity of circulatory myeloid progenitor cells may contribute to enhanced osteoclastogenesis and bone loss in the C57BL/6 mouse model of collagen-induced arthritis

Increased chemotaxis and activity of circulatory myeloid progenitor cells may contribute to enhanced osteoclastogenesis and bone loss in the C57BL/6 mouse model of collagen-induced arthritis

Acute hematopoietic stress in mice is followed by enhanced osteoclast maturation in the bone marrow microenvironment

Recent advances in endotoxin tolerance

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