Expression Of Macrophage Colony-stimulating Factor Research Articles

IL-1α released from damaged epithelial cells is sufficient and essential to trigger inflammatory responses in human lung fibroblasts.

Activation of the innate immune system plays a key role in exacerbations of chronic lung disease, yet the potential role of lung fibroblasts in innate immunity and the identity of epithelial danger signals (alarmins) that may contribute to this process are unclear. The objective of the study was to identify lung epithelial-derived alarmins released during endoplasmic reticulum stress (ER stress) and oxidative stress and evaluate their potential to induce innate immune responses in lung fibroblasts. We found that treatment of primary human lung fibroblasts (PHLFs) with conditioned media from damaged lung epithelial cells significantly upregulated interleukin IL-6, IL-8, monocyte chemotactic protein-1, and granulocyte macrophage colony-stimulating factor expression (P<0.05). This effect was reduced with anti-IL-1α or IL-1Ra but not anti-IL-1β antibody. Costimulation with a Toll-like receptor 3 ligand, polyinosinic–polycytidylic acid (poly I:C), significantly accentuated the IL-1α-induced inflammatory phenotype in PHLFs, and this effect was blocked with inhibitor of nuclear factor kappa-B kinase subunit beta and TGFβ-activated kinase-1 inhibitors. Finally, Il1r1−/− and Il1a−/− mice exhibit reduced bronchoalveolar lavage (BAL) neutrophilia and collagen deposition in response to bleomycin treatment. We conclude that IL-1α plays a pivotal role in triggering proinflammatory responses in fibroblasts and this process is accentuated in the presence of double-stranded RNA. This mechanism may be important in the repeated cycles of injury and exacerbation in chronic lung disease.

Skeletal Actions of Fasting-Induced Adipose Factor (FIAF)

Several adipokines are known to influence skeletal metabolism. Fasting-induced adipose factor (FIAF) is an adipokine that gives rise to 2 further peptides in vivo, the N-terminal coiled-coil domain (FIAF(CCD)) and C-terminal fibrinogen-like domain (FIAF(FLD)). The skeletal action of these peptides is still uncertain. Our results show that FIAF(CCD) is a potent inhibitor of osteoclastogenesis and function, as seen in mouse bone marrow and RAW264.7 cell cultures, and in a resorption assay using isolated primary mature osteoclasts. The inhibitory effects at 500 ng/mL were approximately 90%, 50% and 90%, respectively, in these assays. FIAF(CCD) also stimulated osteoblast mitogenesis by approximately 30% at this concentration. In comparison, FIAF(FLD) was only active in decreasing osteoblast mitogenesis, and intact FIAF had no effect in any of these assays. In murine bone marrow cultures, FIAF(CCD) reduced the expression of macrophage colony-stimulating factor (M-CSF), nuclear factor of activated T-cells c1 (NFATc1) and dendritic cell-specific transmembrane protein (DC-STAMP), and to lesser extent suppressed the expression of connective tissue growth factor (CTGF). FIAF(CCD) also decreased expression of M-CSF and CTGF in stromal/osteoblastic ST2 cells. Its effect on receptor activator of nuclear factor κB (RANKL) and osteoprotegerin expression in bone marrow was not consistent with its inhibitory action on osteoclastogenesis, but it decreased RANKL expression in ST2 cells. In RAW264.7 cell cultures, FIAF(CCD) significantly reduced the expression of NFATc1 and DC-STAMP. In conclusion, FIAF(CCD) inhibits osteoclast differentiation and function in vitro and decreases expression of genes encoding key osteoclastogenic factors such as M-CSF, CTGF, NFATc1, and DC-STAMP. FIAF(CCD)'s action on osteoclasts may be independent of the RANKL/osteoprotegerin pathway. These results suggest a novel mechanism by which adipose tissue may regulate bone resorption and skeletal health.

Abstract B48: Breast tumor cell TACE-shed MCSF promotes pro-angiogenic macrophages through NF-κB signaling

Abstract Mortalities associated with breast cancer, the most common malignancy in women, are mainly due to metastases. Tumor associated macrophages (TAMs) significantly contribute to the development of metastasis through promotion of the rate limiting step of angiogenesis through secretion of vascular endothelial growth factor (VEGF). Macrophage differentiation and migration are activated by macrophage colony stimulating factor (MCSF) and chemokine (C-C motif) ligand 2 (CCL2), respectively. Whereas tumor cells secrete CCL2, MCSF release is mediated by ectodomain shedding through tumor necrosis factor alpha converting enzyme activation (TACE). How tumor cells support macrophage promotion of angiogenesis remains unclear. In the present study, we determined whether MCSF shed by TACE from tumor cells promotes angiogenesis through the activation of NF-κB signaling in macrophages and subsequent VEGF release. These studies were modeled in vitro using a panel of murine mammary cells mimicking breast cancer progression from normal murine mammary gland (NMuMG) cells to metastatic 4T1 cells along with J774 macrophages. TACE and MCSF expressions were higher in metastatic cells compared to epithelial cells (p&amp;lt;0.05). After incubation with 4T1 tumor conditioned media (CM), macrophages secreted significantly more VEGF and expressed of the activated NF-κB, and the resulting macrophage secretions promoted significantly high levels of endothelial cell tubes (p&amp;lt;0.05). Furthermore, a similar activation of macrophages was generated by the combination of CCL2, also highly expressed by tumor cells, and MCSF. These results highlight the key role of tumor cell TACE-shed MCSF and secreted CCL2 in synergistically stimulating pro-angiogenic macrophages. Citation Format: Stephen L. Rego, Rachel Helms, Didier Dreau. Breast tumor cell TACE-shed MCSF promotes pro-angiogenic macrophages through NF-κB signaling. [abstract]. In: Proceedings of the Third AACR International Conference on Frontiers in Basic Cancer Research; Sep 18-22, 2013; National Harbor, MD. Philadelphia (PA): AACR; Cancer Res 2013;73(19 Suppl):Abstract nr B48.

Comparison of the anti-tumor effects of denosumab and zoledronic acid on the neoplastic stromal cells of giant cell tumor of bone

Denosumab and Zoledronic acid (ZOL) are two antiresorptive drugs currently in use for treating osteoporosis. They have different mechanisms of action but both have been shown to delay the onset of skeletal-related events in patients with giant cell tumor of bone (GCT). However, the anti-tumor mechanisms of denosumab on the neoplastic GCT stromal cells remain unknown. In this study, we focused on the direct effects of denosumab on the neoplastic GCT stromal cells and compared with ZOL. The microscopic view demonstrated a reduced cell growth in ZOL-treated but not in denosumab-treated GCT stromal cells. ZOL was found to exhibit a dose-dependent inhibition in cell growth in all GCT stromal cell lines tested and cause apoptosis in two out of three cell lines. In contrast, denosumab only exerted a minimal inhibitory effect in one cell line and did not induce any apoptosis. ZOL significantly inhibited the mRNA expression of receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG) in two GCT stromal cell lines whereas their protein levels remained unchanged. On the contrary, denosumab did not regulate RANKL and OPG expression at both mRNA and protein levels. Moreover, the protein expression of Macrophage Colony-Stimulating Factor (M-CSF), Alkaline Phosphatase (ALP), and Collagen α1 Type I were not regulated by denosumab and ZOL either. Our findings provide new insights in the anti-tumor effect of denosumab on GCT stromal cells and raise a concern that tumor recurrence may occur after the withdrawal of the drug.

Labisia pumila regulates bone-related genes expressions in postmenopausal osteoporosis model.

BackgroundLabisia Pumila var. alata (LPva) has shown potential as an alternative to estrogen replacement therapy (ERT) in prevention of estrogen-deficient osteoporosis. In earlier studies using postmenopausal model, LPva was able to reverse the ovariectomy-induced changes in biochemical markers, bone calcium, bone histomorphometric parameters and biomechanical strength. The mechanism behind these protective effects is unclear but LPva may have regulated factors that regulate bone remodeling. The aim of this study is to determine the bone-protective mechanism of LPva by measuring the expressions of several factors involved in bone formative and resorptive activities namely Osteoprotegerin (OPG), Receptor Activator of Nuclear Factor kappa-B Ligand (RANKL), Macrophage-Colony Stimulating Factor (MCSF) and Bone Morphogenetic Protein-2 (BMP-2).MethodsThirty-two female Wistar rats were randomly divided into four groups: Sham-operated (Sham), ovariectomized control (OVXC), ovariectomized with Labisia pumila var. alata (LPva) and ovariectomized with ERT (Premarin®) (ERT). The LPva and ERT were administered via daily oral gavages at doses of 17.5 mg/kg and 64.5 μg/kg, respectively. Following two months of treatment, the rats were euthanized and the gene expressions of BMP-2, OPG, RANKL and MCSF in the femoral bones were measured using a branch - DNA technique.ResultsThe RANKL gene expression was increased while the OPG and BMP-2 gene expressions were reduced in the OVXC group compared to the SHAM group. There were no significant changes in the MCSF gene expressions among the groups. Treatment with either LPva or ERT was able to prevent these ovariectomy-induced changes in the gene expressions in ovariectomized rats with similar efficacy.ConclusionLPva may protect bone against estrogen deficiency-induced changes by regulating the RANKL, OPG and BMP-2 gene expressions.

Cryopreservation induces macrophage colony stimulating factor from human periodontal ligament cells in vitro

Cryopreservation is used to protect vital periodontal ligaments during the transplantation of teeth. We investigated which gene products implicated in root resorption are upregulated in human periodontal ligament cells by cryopreservation, and whether cryopreservation affects the expression of macrophage-colony stimulating factor (M-CSF) in human periodontal ligament cells. We used customized microarrays to compare gene expression in human periodontal ligament cells cultured from teeth immediately after extraction and from cryopreserved teeth. Based on the result of these assays, we examined M-CSF expression in periodontal ligament cells from the immediately extracted tooth and cryopreserved teeth by real-time PCR, enzyme-linked immunosorbent assay (ELISA), Western blot analysis, and immunofluorescence. We also investigated whether human bone marrow cells differentiate into tartrate-resistant acid phosphatase (TRAP) positive osteoclasts when stimulated with RANKL (Receptor Activator for Nuclear Factor κ B Ligand) together with any secreted M-CSF present in the supernatants of the periodontal ligament cells cultured from the various groups of teeth. M-CSF was twofold higher in the periodontal ligament cells from the rapid freezing teeth than in those from the immediately extracted group (p<0.05). Cryopreservation increased M-CSF expression in the periodontal ligament cells when analyzed by real time PCR, ELISA, Western blotting, and immunofluorescence (p<0.05). TRAP positive osteoclasts were formed in response to RANKL and the secreted M-CSF present in the supernatants of all the experimental groups except negative control. These results demonstrate that cryopreservation promotes the production of M-CSF, which plays an important role in root resorption by periodontal ligament cells.

Pro-inflammatory and anti-angiogenic effects of bisphosphonates on human cultured retinal pigment epithelial cells

AimBisphosphonates have been shown to induce ocular inflammatory diseases such as uveitis and scleritis, while being protective against angiogenic diseases like neovascular age-related macular degeneration (AMD). Therefore, we studied the...

Abstract 1424: Inhibition of M-CSF or MCP-1 expression in tumor cells reduces tumor growth and lung metastasis in a murine mammary cancer model.

Abstract Tumor-associated macrophages, which are derived from the infiltration of circulating bone marrow-derived monocytes, consist primarily of a polarized M2 macrophage population and are associated with poor prognosis in various cancers. We previously observed that solid tumor growth and lung metastasis were enhanced when 4T1 mammary carcinoma cells were injected with M2 macrophages into the mammary fat pad of BALB/c mice. Cancer cell proliferation and macrophage infiltration were markedly increased when M2 macrophages were co-injected with 4T1 cells. In the present study, we explored the mechanisms underlying the crosstalk between 4T1 cells and M2 macrophages in tumor microenvironment. The results of in vitro experiments revealed that the migration of monocytes and the secretion of many cytokines including macrophage-colony stimulating factor (M-CSF) and monocyte chemotactic protein-1 (MCP-1) were increased when 4T1 cells were co-cultured with M2 macrophages, as compared to when the 4T1 cells were cultured alone. The mRNA expression of M-CSF and MCP-1 increased in 4T1 cells when 4T1 cells were co-cultured with M2 macrophages. To investigate the potential roles of M-CSF and MCP-1 in tumor growth and metastasis, 4T1 cells were infected with M-CSF, MCP-1, or control shRNA lentivirus particles and selected with puromycin. 4T1 cells infected with M-CSF shRNA did not grow or metastasize to the lung when injected into the mammary fat pads of BALB/c mice. Inhibition of MCP-1 expression in tumor cells also suppressed 4T1 tumor growth and metastasis, however, the degree of inhibition was much smaller than that of M-CSF inhibition. M-CSF knockdown decreased M2 macrophage (mouse mannose receptor+) population, whereas MCP-1 knockdown decreased the number of both M1 and M2 macrophages in tumor tissues. Taken together, these results indicate that M-CSF and MCP-1 are produced by the crosstalk between 4T1 cells and M2 macrophages, and these cytokines play important roles in mammary tumor progression. Citation Format: Han Jin Cho, Jung Han Yoon Park. Inhibition of M-CSF or MCP-1 expression in tumor cells reduces tumor growth and lung metastasis in a murine mammary cancer model. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1424. doi:10.1158/1538-7445.AM2013-1424

Suppressed expression of granulocyte macrophage colony-stimulating factor in oviduct ampullae of obese cows

Obese heifers have been found to produce fewer excellent-grade embryos than lean and normal heifers due to unknown mechanisms. Oviducts synthesize granulocyte macrophage colony-stimulating factor (GMCSF) to promote embryogenesis, and GMCSF expression may be down-regulated in the oviducts of obese cows. The present study evaluated the relationship between the degree of obesity and GMCSF expression in the ampullary or isthmic section of oviducts in lean [n=5; body condition score (BCS) on a 5-point scale, 2.5], normal (n=6; BCS, 3.0), and obese (n=5; BCS, 4.0) Japanese Black cows. GMCSF mRNA and protein expression in the ampulla, measured by real-time PCR and western blotting, respectively, were less (P<0.05) in the obese group than in the normal group. mRNA and GMCSF protein did not differ significantly in the isthmus among the three groups. The obese group had less GMCSF immuno-reactivity in the tunica mucosa, the primary site of GMCSF gene expression, of the ampulla than the normal and lean groups. In conclusion, unlike normal and lean cows, obese cows had suppressed GMCSF gene expression in the ampulla.

Compressive stress induced the up-regulation of M-CSF, RANKL, TNF-α expression and the down-regulation of OPG expression in PDL cells via the integrin-FAK pathway

ObjectivesThis study was performed to elucidate the involvement of integrin-FAK (focal adhesion kinase) pathway in compressive stress-induced mRNA expression of macrophage colony stimulating factor (M-CSF), tumour necrosis factor (TNF)-α, receptor activator of nuclear factor κB (RANKL) and osteoprotegerin (OPG) and to further confirm the role of the integrin-FAK complex as a mechanoreceptor in PDL cells. DesignPeriodontal ligament (PDL) cells were obtained from patients having healthy first premolars extracted for orthodontic purposes. Cultured PDL cells were divided into three groups: the control group in which compressive stress was administered; the negative control group in which mechanical stress was administered after transfection of negative control siRNA; and FAK knockdown group in which mechanical stress was administered after FAK siRNA treatment. Compressive stress (2g/cm2) was for various time durations (0.5, 2, 6, 24, 48h). Total RNA was collected after the experiment and real-time PCR analysis was performed to determine the mRNA expression levels of M-CSF, TNF-α, RANKL and OPG. Also the supernatant was analysed with ELISA to detect the corresponding cytokine concentrations. ResultsThe cells of the control group and the negative control group expressed higher mRNA levels of M-CSF, TNF-α, and RANKL but a lower mRNA level of OPG compared to those of baseline. FAK knockdown cells showed lower mRNA expression levels of M-CSF, TNF-α, and RANKL but a higher mRNA expression level of OPG than that in the control. The OPG mRNA expression level in FAK knockdown cells was even higher than that of baseline. ELISA results showed similar pattern of cytokine concentration changes. ConclusionsResults of this study indicate that the integrin-FAK pathway regulates compressive stress-induced expression of M-CSF, TNF-α, RANKL and OPG and suggests that the integrin-FAK complex acts as a mechanoreceptor in PDL cells.

The expression and molecular mechanism of M1 macrophages in rheumatic valvular disease

To investigate whether valvular expression of macrophage and its subsets and correlative cytokines of mitral valve are altered in patients with rheumatic valvular disease. The mitral valvular leaflets of 15 rheumatic valvular disease patients were included as the test group, and 7 patients of terminal stage cardiomyopathy as the control. The immunostain of CD68, inducible nitric oxide synthase (iNOS) and CD163 were applied to mark the total macrophages, M1 and M2, respectively. The expression of endothelial nitric oxide synthase (eNOS), IL-10, Arg-1, macrophage-colony stimulating factor (M-CSF) were compared respectively in two groups. The angiogenesis was enormous in the test, whereas the cell proliferation was scanty. Compared with the control, CD68 positive macrophages were markly expressed in the test (4.2 ± 2.0 vs. 3.2 ± 2.3; Z = -3.981, P = 0.000), also the iNOS positive M1 subsets (3.4 ± 1.7 vs. 1.2 ± 1.0; Z = -4.015, P = 0.000). The expression level of CD163 positive macrophages was lower in the test (1.2 ± 1.0 vs. 2.3 ± 1.8; Z = -8.602, P = 0.000). The expression of eNOS was higher in the rheumatic valve disease (4.9 ± 1.1 vs. 1.8 ± 1.1), but the expression levels of Arg-1 (1.0 ± 1.0 vs. 3.3 ± 1.3) and IL-10 (2.1 ± 1.2 vs. 4.9 ± 1.4) were lower (Z = -8.867 to -5.344, P = 0.000). The expression of M-CSF was lower in test (2.0 ± 1.4 vs. 4.3 ± 0.9; Z = -2.741, P = 0.006). The infiltration of M1 macrophages plays an important role in the progression of rheumatic mitral valve disease. It fulfills the pro-inflammation by up-regulating the expression of eNOS. Inversely, it suppresses the expression of IL-10, Arg-1 to relieve the inflammatory action. In according with the down-regulated level of M-CSF, the polarization from M1 macrophages into M2 is depressed, and the inflammation induced by M1 is sustained.

Eurycoma longifolia upregulates osteoprotegerin gene expression in androgen- deficient osteoporosis rat model

BackgroundEurycoma longifolia (EL) has been shown recently to protect against bone calcium loss in orchidectomised rats, the model for androgen-deficient osteoporosis. The mechanism behind this is unclear but it may be related to its ability to elevate testosterone levels or it may directly affect bone remodeling. The aim of this study is to determine the mechanism involved by investigating the effects of EL extract on serum testosterone levels, bone biomarkers, biomechanical strength and gene expression of Receptor Activator of Nuclear Factor kappa-B ligand (RANKL), Osteoprotegerin (OPG) and Macrophage-Colony Stimulating Factor (MCSF) in orchidectomised rats.MethodsThirty-two male Sprague–Dawley rats were divided into: Sham-operated group (SHAM); orchidectomised-control group (ORX); orchidectomised and given 15 mg/kg EL extract (ORX + EL) and orchidectomised and given 8 mg/kg testosterone (ORX + T). The rats were treated for 6 weeks. The serum levels of testosterone, osteocalcin and C-terminal telopeptide of type I collagen (CTX) were measured using the ELISA technique. The femoral bones were subjected to biomechanical testing. The tibial bone gene expressions of RANKL, OPG and MCSF were measured using the branch DNA technique.ResultsThe post-treatment level of testosterone was found to be significantly reduced by orchiectomy (p < 0.05). Both ORX + EL and ORX + T groups have significantly higher post-treatment testosterone levels compared to their pre-treatment levels (p < 0.05). The bone resorption marker (CTx) was elevated after orchiectomy but was suppressed after treatment in the ORX + EL and ORX + T groups (p < 0.05). There was no significant finding for the femoral biomechanical parameters. The tibial OPG gene expression in the ORX group was significantly lower compared to the SHAM and ORX + EL groups (p < 0.05).ConclusionSupplementation with EL extract elevated the testosterone levels, reduced the bone resorption marker and upregulated OPG gene expression of the orchidectomised rats. These actions may be responsible for the protective effects of EL extract against bone resorption due to androgen deficiency.

Effects of type 2 diabetes mellitus on expression of macrophage colony-stimulating factor and bone differentiation factors in human chronic periodontitis

Effects of type 2 diabetes mellitus on expression of macrophage colony-stimulating factor and bone differentiation factors in human chronic periodontitis

Augmented Macrophage Differentiation and Polarization of Tumor-associated Macrophages Towards M1 Subtype in Listeria-administered Tumor-bearing Host

This study investigates the effect of Listeria administration on differentiation of macrophages from precursor bone marrow cells and functional status of tumor-associated macrophages (TAM). Listeria administration not only resulted in an augmented infiltration of tumor by F4/80 macrophages but also repolarized the functional status of TAM displaying features of some M1 macrophage subtype with upregulated phagocytosis and tumoricidal activity accompanied by altered expression of monocarboxylate transporter-1, toll-like receptor-2, surface markers: CD11c, interleukin-2 receptor, CD62L, and secreted molecules: nitric oxide, interleukin (IL)-1, IL-6, tumor necrosis factor-α, and vascular endothelial growth factor. Declined tumor cell survival and modulated repertoire of cytokines: interferon-γ, IL-6, IL-10, and transforming growth factor-β in tumor microenvironment indicated their role in polarization of TAM towards proinflammatory state. Bone marrow cell of Listeria-administered tumor-bearing mice showed augmented survival, declined expression of p53 upregulated modulator of apoptosis with an upregulated differentiation into activation responsive bone marrow-derived macrophages along with altered expression of macrophage-colony stimulating factor, macrophage-colony stimulating factor receptor, and granulocyte macrophage-colony stimulating factor receptor. These findings indicate that Listeria infection is associated with an augmented differentiation of macrophages accompanied by tumoricidal activation of TAM.

Osteoblasts stimulate osteoclastogenesis via RANKL expression more strongly than periodontal ligament cells do in response to PGE2

ObjectivePeriodontal ligament cells (PDLs) produce prostaglandin E2 (PGE2) in response to orthodontic force. PGE2 is a potent osteoclast-inducing factor that induces the receptor activator of nuclear factor-κB ligand (RANKL). Some studies reported that PDLs express RANKL in response to mechanical stress, whereas another study reported that they do not. Based on an immunohistochemical study, RANKL expression is localized around the alveolar bone surface 3 days after tooth movement. However, ankylosed teeth cannot be moved by therapeutic mechanical stress, suggesting that PDLs play a major role in alveolar bone resorption. In this study, we compared the functional difference in osteoclastogenesis between human PDLs (HPDLs) and normal human osteoblasts (HOBs) as a direct effect of PGE2 exposure. DesignWe examined the expression of RANKL, osteoprotegerin, and macrophage colony-stimulating factor after 48-h culture with or without PGE2 (10−11 to 10−5M) in HPDLs and HOBs. Then to confirm whether RANKL produced by PGE2 treatment induces osteoclastogenesis or not, RAW264.7 cells were co-cultured on HPDLs or HOBs pretreated with 10−6M of PGE2. ResultPGE2 exposure increased significantly RANKL expression in HOBs compared with HPDLs. PGE2 exposure significantly decreased osteoprotegerin expression in HPDLs compared with HOBs. The number of tartrate-resistant acid phosphatase staining osteoclast-like cells from RAW264.7 cells increased significantly by PGE2 pretreatment in HOBs and was reduced by small interfering RNA knockdown of RANKL. ConclusionThese results suggest that osteoblasts strongly influence the stimulation of osteoclastogenesis via RANKL, induced by PGE2 in periodontal tissues, compared with PDLs.

Increased expression of interleukin-1β in triglyceride-induced macrophage cell death is mediated by p38 MAP kinase

Triglycerides (TG) are implicated in the development of atherosclerosis through formation of foam cells and induction of macrophage cell death. In this study, we report that addition of exogenous TG induced cell death in phorbol 12-myristate 13-acetate-differentiated THP-1 human macrophages. TG treatment induced a dramatic decrease in interleukin-1β (IL-1β) mRNA expression in a dose- and time-dependent manner. The expression of granulocyte macrophage colony-stimulating factor and platelet endothelial cell adhesion molecule remained unchanged. To identify signaling pathways involved in TG-induced downregulation of IL-1β, we added p38 MAPK, protein kinase C (PKC) or c-Raf1 specific inhibitors. We found that inhibition of p38 MAPK alleviated the TG-induced downregulation of IL-1β, whereas inhibition of PKC and c-Raf1 had no effect. This is the first report showing decreased IL-1β expression during TG-induced cell death in a human macrophage line. Our results suggest that downregulation of IL-1β expression by TG-treated macrophages may play a role during atherogenesis.

An Overview of Molecular and Cellular Mechanisms Associated with Porcine Pregnancy Success or Failure

Prenatal mortality remains one of the major constraints for the commercial pig industry in North America. Twenty to thirty per cent of the conceptuses are lost early in gestation and an additional 10-15% is lost by mid-to-late gestation. Research over the last two decades has provided critical insights into how uterine capacity, placental efficiency, genetics, environment, nutrition and immune mechanisms impact successful conceptus growth; however, the exact cause and effect relationship in the context of foetal loss has yet to be determined. Similar to other mammalian species such as the human, mouse, rat, and primates, immune cell enrichment occurs at the porcine maternal-foetal interface during the window of conceptus attachment. However, unlike other species, immune cells are solely recruited by conceptus-derived signals. As pigs have epitheliochorial placentae where maternal and foetal tissue layers are separate, it provides an ideal model to study immune cell interactions with foetal trophoblasts. Our research is focused on the immune-angiogenesis axis during porcine pregnancy. It is well established that immune cells are recruited to the maternal-foetal interface, but their pregnancy specific functions and how the local milieu affects angiogenesis and inflammation at the site of foetal arrest remain unknown. Through a better understanding of how immune cells modulate crosstalk between the conceptus and the mother, it might be possible to therapeutically target immune cells and/or their products to reduce foetal loss. In this review, we provide evidence from the literature and from our own work into the immunological factors associated with porcine foetal loss.

Inflammatory protein profiling of pancreatic cyst fluid using EUS-FNA in tandem with cytokine microarray differentiates between branch duct IPMN and inflammatory cysts

BackgroundDiagnosis of pancreatic cystic neoplasms remains problematic. We hypothesize that inflammatory mediator proteins in pancreatic cyst fluid can differentiate branch duct intraductal papillary mucinous neoplasms (BD-IPMNs) and pancreatic inflammatory cysts. We aim to 1) detect inflammatory mediator proteins (IMPs) using a multiplexed IMP-targeted microarray in pancreatic cyst fluid obtained during endoscopic ultrasound fine needle aspiration (EUS-FNA) and 2) compare IMP profiles in pancreatic cyst fluid from BD-IPMNs and inflammatory cysts. Pancreatic cyst fluid from ten patients (5 BD-IPMN and 5 inflammatory cysts) was obtained by EUS-FNA and analyzed directly with a multiplexed microarray assay to determine concentrations of 89 IMPs. Statistical analysis was performed using non-parametric methods. ResultsEighty-three of the 89 assayed IMPs were detected in at least one of the 10 patient samples. Seven IMPs were detected in BD-IPMN but not inflammatory cysts, while eleven IMPs were identified in inflammatory cysts but not BD-IPMN. Notably, granulocyte–macrophage colony-stimulating factor (GM-CSF) expression was present in all five inflammatory cyst samples. Hepatocyte growth factor (HGF) was present in significantly higher concentrations in inflammatory cysts compared to BD-IPMN. ConclusionOur exploratory analysis reveals that GM-CSF and HGF in EUS-FNA-collected pancreatic cyst fluid can distinguish between BD-IPMN and inflammatory cyst. Coupling microarray molecular techniques to EUS-FNA may represent a major step forward to our understanding complex pancreatic disease.

Bisphosphonate- and statin-induced enhancement of OPG expression and inhibition of CD9, M-CSF, and RANKL expressions via inhibition of the Ras/MEK/ERK pathway and activation of p38MAPK in mouse bone marrow stromal cell line ST2

Osteoclast differentiation is influenced by receptor activator of the NF-κB ligand (RANKL), macrophage colony-stimulating factor (M-CSF), and CD9, which are expressed on bone marrow stromal cells and osteoblasts. In addition, osteoprotegerin (OPG) is known as an osteoclastogenesis inhibitory factor. In this study, we investigated whether bisphosphonates and statins increase OPG expression and inhibit the expression of CD9, M-CSF, and RANKL in the bone marrow-derived stromal cell line ST2. We found that bisphosphonates and statins enhanced OPG mRNA expression and inhibited the expression of CD9, M-CSF, and RANKL mRNA. Futhermore, bisphosphonates and statins decreased the membrane localization of Ras and phosphorylated ERK1/2, and activated the p38MAPK. This indicates that bisphosphonates and statins enhanced OPG expression, and inhibited the expression of CD9, M-CSF, and RANKL through blocking the Ras/ERK pathway and activating p38MAPK. Accordingly, we believe that its clinical applications will be investigated in the future for the development of osteoporosis therapy.

Prognostic impact of CD57, CD68, M-CSF, CSF-1R, Ki67 and TGF-beta in soft tissue sarcomas

BackgroundPrognostic markers in curable STS may have the potential to guide therapy after surgical resection. The purpose of this study was to clarify the prognostic impact of the presence of cells and growth factors belonging to the innate immune system in soft tissue sarcomas (STS). The significance of macrophages (CD68), their growth factor macrophage colony-stimulating factor (M-CSF), its receptor colony-stimulating factor-1 receptor (CSF-1R), natural killer cells (CD57) and the general immunomodulating molecule (TGF-beta) are all controversial in STS. Herein, these markers are evaluated and compared to the cell proliferation marker Ki67.MethodsTissue microarrays from 249 patients with non-gastrointestinal (non-GIST) STS were constructed from duplicate cores of viable and representative neoplastic tumor areas and duplicate cores of peritumoral capsule. Immunohistochemistry was used to evaluate the expression of CD68, M-CSF, CSF-1R, CD57, TGF-beta and Ki67 in tumor and peritumoral capsule.ResultsIn univariate analyses increased expression of M-CSF (P = 0.034), Ki67 (P < 0.001) and TGF-beta (P = 0.003) in tumor correlated with shorter disease-specific survival (DSS). Increased expression of CD68 in tumor correlated significantly with malignancy grade (P = 0.016), but not DSS (P = 0.270). Increased expression of Ki67 in peritumoral capsule tended to correlate with a shorter DSS (P = 0.057). In multivariate analyses, co-expression of M-CSF and TGF-beta (P = 0.022) in tumor and high expression of Ki67 (P = 0.019) in peritumoral capsule were independent negative prognostic factors for DSS.ConclusionsIncreased co-expression of M-CSF and TGF-beta in tumor in patients with STS, and increased expression of Ki67 in peritumoral capsule were independent negative prognostic factors for DSS.