Browsing by Subject "macrophage"

Now showing 1 - 6 of 6
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    Association of Plasma CD163 Concentration with De Novo–Onset Chronic Graft-versus-Host Disease
    (Elsevier, 2017) Inamoto, Yoshihiro; Martin, Paul J.; Paczesny, Sophie; Tabellini, Laura; Momin, Amin A.; Mumaw, Christen L.; Flowers, Mary E. D.; Lee, Stephanie J.; Carpenter, Paul A.; Storer, Barry E.; Hanash, Samir; Hansen, John A.; Department of Pediatrics, IU School of Medicine
    Chronic graft-versus-host disease (GVHD) is the leading cause of long-term morbidity and mortality after allogeneic hematopoietic cell transplantation. To identify prognostic plasma proteins associated with de novo– or quiescent-onset chronic GVHD (cGVHD), we performed a discovery and validation proteomic study. The total study cohort included 167 consecutive patients who had no clinical evidence of GVHD under minimum glucocorticoid administration and had available plasma samples obtained at 80 ± 14 days after transplantation. We first used high-throughput mass spectrometry to screen pooled plasma using 20 cases with subsequent cGVHD and 20 controls without it, and we identified 20 candidate proteins. We then measured 12 of the 20 candidate proteins by ELISA on the same individual samples and identified 4 proteins for further verification (LGALS3BP, CD5L, CD163, and TXN for de novo onset, and LGALS3BP and CD5L for quiescent onset). The verification cohort included 127 remaining patients. The cumulative incidence of de novo–onset cGVHD was higher in patients with higher plasma soluble CD163 concentrations at day 80 than those with lower concentrations (75% versus 40%, P = .018). The cumulative incidence of de novo– or quiescent-onset cGVHD did not differ statistically according to concentrations of the 3 other proteins at day 80. CD163 is a macrophage scavenger receptor and is elevated in oxidative conditions. These results suggest that monocyte or macrophage activation or increased oxidative stress may contribute to the pathogenesis of cGVHD.
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    CD68 Macrophage Expression in Normal Breast Tissue and Cancer
    (2019-04-19) Gaines, Madelynn; Jacobsen, Max; Temm, Connie; Sandusky, George
    Breast cancer is a common disease and is the second leading cause of death in women. This type of cancer is usually hormonally driven by estrogen, progesterone, and HER2. Macrophages play a large role in the tumor microenvironment (TME). The aim of this study was to investigate the percent of macrophages in 32 normal breast tissues, 66 normal adjacent tissue (NAT), and 82 breast cancer tissues using the CD68-specific biomarker. Tissue microarrays (TMA) were created, which are composed of 2-mm cores from multiple patients mounted onto a single slide. The breast tissue samples were fixed, processed, microtomed, and stained with CD68. Unstained slides were immunostained using the Dako FLEX system. The slides were imaged using the Aperio Whole Slide Imaging platform and the tissues were evaluated using the positive pixel count algorithm (a quantitative image analysis system). The positivity of macrophages in the tissue samples were reported as a percentage, and compared across the three groups. It was found that the CD68 positivity in the normal and breast cancer tissue were even, and the NAT was lower. However, the three groups had overlapping standard deviations. Because difference between the percentages of each group was minimal and the deviations overlapped, it was concluded that there is no statistical difference between the three groups.
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    Myo-Inositol in Fermented Sugar Matrix Improves Human Macrophage Function
    (Wiley, 2022) Ghosh, Nandini; Das, Amitava; Biswas, Nirupam; Mahajan, Sanskruti P.; Madeshiya, Amit K.; Khanna, Savita; Sen, Chandan K.; Roy, Sashwati; Surgery, School of Medicine
    Scope Reactive oxygen species production by innate immune cells plays a central role in host defense against invading pathogens at wound-site. A weakened hos-defense results in persistent infection leading to wound chronicity. Fermented Papaya Preparation (FPP), a complex sugar matrix, bolstered respiratory burst activity and improved wound healing outcomes in chronic wound patients. The objective of the current study was to identify underlying molecular factor/s responsible for augmenting macrophage host defense mechanisms following FPP supplementation. Methods and results In depth LC-MS/MS analysis of cells supplemented with FPP led to identification of myo-inositol as a key determinant of FPP activity towards improving macrophage function. Myo-inositol, in quantities that is present in FPP, significantly improved macrophage respiratory burst and phagocytosis via de novo synthesis pathway of ISYNA1. Additionally, myo-inositol transporters, HMIT and SMIT1, played a significant role in such activity. Blocking these pathways using siRNA attenuated FPP-induced improved macrophage host defense activities. FPP supplementation emerges as a novel approach to increase intracellular myo-inositol levels. Such supplementation also modified wound microenvironment in chronic wound patients to augment myo-inositol levels in wound fluid. Conclusion These observations indicate that myo-inositol in FPP influences multiple aspects of macrophage function critical for host defense against invading pathogens.
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    Polarization of Macrophages toward M2 Phenotype Is Favored by Reduction in iPLA2β (Group VIA Phospholipase A2)
    (American Society for Biochemistry and Molecular Biology, 2016-10-28) Ashley, Jason W.; Hancock, William D.; Nelson, Alexander J.; Bone, Robert N.; Tse, Hubert M.; Wohltmann, Mary; Turk, John; Ramanadham, Sasanka; Medicine, School of Medicine
    Macrophages are important in innate and adaptive immunity. Macrophage participation in inflammation or tissue repair is directed by various extracellular signals and mediated by multiple intracellular pathways. Activation of group VIA phospholipase A2 (iPLA2β) causes accumulation of arachidonic acid, lysophospholipids, and eicosanoids that can promote inflammation and pathologic states. We examined the role of iPLA2β in peritoneal macrophage immune function by comparing wild type (WT) and iPLA2β−/− mouse macrophages. Compared with WT, iPLA2β−/− macrophages exhibited reduced proinflammatory M1 markers when classically activated. In contrast, anti-inflammatory M2 markers were elevated under naïve conditions and induced to higher levels by alternative activation in iPLA2β−/− macrophages compared with WT. Induction of eicosanoid (12-lipoxygenase (12-LO) and cyclooxygenase 2 (COX2))- and reactive oxygen species (NADPH oxidase 4 (NOX4))-generating enzymes by classical activation pathways was also blunted in iPLA2β−/− macrophages compared with WT. The effects of inhibitors of iPLA2β, COX2, or 12-LO to reduce M1 polarization were greater than those to enhance M2 polarization. Certain lipids (lysophosphatidylcholine, lysophosphatidic acid, and prostaglandin E2) recapitulated M1 phenotype in iPLA2β−/− macrophages, but none tested promoted M2 phenotype. These findings suggest that (a) lipids generated by iPLA2β and subsequently oxidized by cyclooxygenase and 12-LO favor macrophage inflammatory M1 polarization, and (b) the absence of iPLA2β promotes macrophage M2 polarization. Reducing macrophage iPLA2β activity and thereby attenuating macrophage M1 polarization might cause a shift from an inflammatory to a recovery/repair milieu.
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    Protein-tyrosine Phosphatase Shp2 Positively Regulates Macrophage Oxidative Burst
    (2015-02) Li, Xing Jun; Goodwin, Charles B.; Nabinger, Sarah C.; Richine, Briana M.; Yang, Zhenyun; Hanenberg, Helmut; Ohnishi, Hiroshi; Matozaki, Takashi; Feng, Gen-Sheng; Chan, Rebecca J.; Department of Pediatrics, Indiana University School of Medicine
    Macrophages are vital to innate immunity and express pattern recognition receptors and integrins for the rapid detection of invading pathogens. Stimulation of Dectin-1 and complement receptor 3 (CR3) activates Erk- and Akt-dependent production of reactive oxygen species (ROS). Shp2, a protein-tyrosine phosphatase encoded by Ptpn11, promotes activation of Ras-Erk and PI3K-Akt and is crucial for hematopoietic cell function; however, no studies have examined Shp2 function in particulate-stimulated ROS production. Maximal Dectin-1-stimulated ROS production corresponded kinetically to maximal Shp2 and Erk phosphorylation. Bone marrow-derived macrophages (BMMs) from mice with a conditionally deleted allele of Ptpn11 (Shp2flox/flox;Mx1Cre+) produced significantly lower ROS levels compared with control BMMs. Although YFP-tagged phosphatase dead Shp2-C463A was strongly recruited to the early phagosome, its expression inhibited Dectin-1- and CR3-stimulated phospho-Erk and ROS levels, placing Shp2 phosphatase function and Erk activation upstream of ROS production. Further, BMMs expressing gain of function Shp2-D61Y or Shp2-E76K and peritoneal exudate macrophages from Shp2D61Y/+;Mx1Cre+ mice produced significantly elevated levels of Dectin-1- and CR3-stimulated ROS, which was reduced by pharmacologic inhibition of Erk. SIRPα (signal regulatory protein α) is a myeloid inhibitory immunoreceptor that requires tyrosine phosphorylation to exert its inhibitory effect. YFP-Shp2C463A-expressing cells have elevated phospho-SIRPα levels and an increased Shp2-SIRPα interaction compared with YFP-WT Shp2-expressing cells. Collectively, these findings indicate that Shp2 phosphatase function positively regulates Dectin-1- and CR3-stimulated ROS production in macrophages by dephosphorylating and thus mitigating the inhibitory function of SIRPα and by promoting Erk activation.
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    SENP1 regulates IFN-γ−STAT1 signaling through STAT3−SOCS3 negative feedback loop
    (Oxford, 2017-04) Yu, Tingting; Zuo, Yong; Cai, Rong; Huang, Xian; Wu, Shuai; Zhang, Chenxi; Chin, Y. Eugene; Li, Dongdong; Zhang, Zhenning; Xia, Nansong; Wang, Qi; Shen, Hao; Yao, Xuebiao; Zhang, Zhong-Yin; Xue, Song; Shen, Lei; Cheng, Jinke; Biochemistry and Molecular Biology, School of Medicine
    Interferon-γ (IFN-γ) triggers macrophage for inflammation response by activating the intracellular JAK−STAT1 signaling. Suppressor of cytokine signaling 1 (SOCS1) and protein tyrosine phosphatases can negatively modulate IFN-γ signaling. Here, we identify a novel negative feedback loop mediated by STAT3−SOCS3, which is tightly controlled by SENP1 via de-SUMOylation of protein tyrosine phosphatase 1B (PTP1B), in IFN-γ signaling. SENP1-deficient macrophages show defects in IFN-γ signaling and M1 macrophage activation. PTP1B in SENP1-deficient macrophages is highly SUMOylated, which reduces PTP1B-induced de-phosphorylation of STAT3. Activated STAT3 then suppresses STAT1 activation via SOCS3 induction in SENP1-deficient macrophages. Accordingly, SENP1-deficient macrophages show reduced ability to resist Listeria monocytogenes infection. These results reveal a crucial role of SENP1-controlled STAT1 and STAT3 balance in macrophage polarization.