ABSTRACT
BACKGROUND: A 70-year-old man with hepatitis C virus-related recurrent hepatocellular carcinoma was admitted for further diagnosis of a 1 cm iso-hyperechoic nodule in segment (S) 5. CASE SUMMARY: Gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid-enhanced magnetic resonance imaging (EOB-MRI) revealed the nodule in S5 with a defect at the hepatobiliary phase, hyperintensity on diffusion weighted imaging (DWI) and hypointensity on apparent diffusion coefficient (ADC) map. Contrast-enhanced computed tomography revealed hypervascularity at the early phase, and delayed contrast-enhancement was observed at the late phase. Contrast-enhanced ultrasound (US) revealed incomplete defect at the late vascular phase. Inflammatory liver tumor, lymphoproliferative disease, intrahepatic cholangiocarcinoma (small duct type) and bile duct adenoma were suspected through the imaging studies. US guided biopsy, however, showed a noncaseating hepatic sarcoid-like epithelioid granuloma (HSEG), and histopathological analysis disclosed spindle shaped epithelioid cells harboring Langhans-type multinucleated giant cells. One month after admission, EOB-MRI signaled the disappearance of the defect at the hepatobiliary phase, of hyperintensity on DWI, of hypointensity on ADC map, and no stain at the early phase. CONCLUSION: That the patient had received BNT162b2 messenger RNA (mRNA) coronavirus disease 2019 vaccination 3 mo before the occurrence of HSEG, and that its disappearance was confirmed 4 mo after mRNA vaccination suggested that the drug-induced sarcoidosis-like reaction (DISR) might be induced by the mRNA vaccination. Fortunately, rechallenge of drug-induced DISR with the third mRNA vaccination was not confirmed.
ABSTRACT
The transcription factor MafB is specifically expressed in macrophages. We have recently demonstrated that MafB is expressed in anti-inflammatory alternatively activated M2 macrophages in vitro. Tumor-associated macrophages (TAMs) are a subset of M2 type macrophages that can promote immunosuppressive activity, induce angiogenesis, and promote tumor cell proliferation. To examine whether MafB express in TAMs, we analyzed green fluorescent protein (GFP) expression in Lewis lung carcinoma tumors of MafB-GFP knock-in heterozygous mice. FACS analysis demonstrated GFP fluorescence in cells positive for macrophage-markers (F4/80, CD11b, CD68, and CD204). Moreover, quantitative RT-PCR analysis with F4/80+GFP+ and F4/80+GFP- sorted cells showed that the GFP-positive macrophages express IL-10, Arg-1, and TNF-α, which were known to be expressed in TAMs. These results indicate that MafB is expressed in TAMs. Furthermore, immunostaining analysis using an anti-MAFB antibody revealed that MAFB is expressed in CD204-and CD68-positive macrophages in human lung cancer samples. In conclusion, MafB can be a suitable marker of TAMs in both mouse and human tumor tissues.
Subject(s)
Carcinoma, Lewis Lung/pathology , Lung Neoplasms/pathology , Macrophages/pathology , MafB Transcription Factor/analysis , Animals , Biomarkers, Tumor/analysis , Cell Line, Tumor , Humans , Mice , Mice, Inbred C57BL , Tumor MicroenvironmentABSTRACT
Butyric acid, a short-chain fatty acid and one of the main metabolites of intestinal microbial fermentation of dietary fiber, has been shown to have an important role in maintaining the integrity of the intestinal mucosa, while it also has been shown to exert potent anti-inflammatory effects both in vitro and in vivo. However, the precise mechanisms underlying those effects have not been fully identified. We exposed colonic epithelial cells to butyric acid, then extracted total RNA samples, and subsequently hybridized them to microarray chips. Among the upregulated genes, milk fat globule-epidermal growth factor 8 (MFG-E8) was elevated by approximately fivefold. We previously reported that the potential therapeutic benefits of MFG-E8 in intestinal tissue injury were dependent not only on enhanced clearance of apoptotic cells but also required diverse cellular events for maintaining epithelial integrity. The influence of butyric acid on cell function is often attributed to its inhibition of histone deacetylases (HDACs). We found that acetylation on histone 3 lysine 9 (acetyl-H3K9) around the MFG-E8 promoter was significantly increased with butyric acid exposure. Experimental colitis was induced by administration of dextran sodium sulfate (DSS) in C57BL/6N (MFG-E8+/+) and MFG-E8-/- mice. Although the colonic bacterial compositions in wild-type (WT) and MFG-E8-/- mice were not significantly different, intrarectal administration of butyric acid during an acute phase of colitis attenuated intestinal inflammatory parameters and inhibited body weight loss in the WT mice. Our novel findings suggest that butyric acid has significant anti-inflammatory effects partly via MFG-E8 on DSS-induced murine experimental colitis.
Subject(s)
Antigens, Surface/metabolism , Butyric Acid/therapeutic use , Colitis/drug therapy , Histamine Antagonists/therapeutic use , Milk Proteins/metabolism , Administration, Rectal , Animals , Butyric Acid/pharmacology , Cell Line , Colitis/chemically induced , Colitis/metabolism , Colitis/microbiology , Dextran Sulfate , Disease Models, Animal , Histamine Antagonists/pharmacology , Histones/metabolism , Male , Metagenome/drug effects , Mice , Mice, Inbred C57BL , Mice, Knockout , Promoter Regions, Genetic , Up-RegulationABSTRACT
Milk fat globule epidermal growth factor-8 (MFG-E8) promotes phagocytic clearance of apoptotic cells to maintain normal tissue homeostasis. However, its functions in intestinal inflammatory disorders are unknown. Since the pathogenesis of those disorders are due to abnormal interactions between intestinal epithelial cells (IECs) and microbial pathogens, we analyzed the effects of MFG-E8 on IECs to determine its protective role in murine experimental colitis. Expression of αvß3-integrin in Colon-26 cells was examined by RT-PCR and immunostaining. Colon-26 cells were pretreated with recombinant wild-type and mutant MFG-E8 proteins, following stimulation with flagellin as an inducer of innate immunity, and the effects of the recombinant proteins on inhibition of nuclear factor-κB (NF-κB) and inflammatory cytokine production in flagellin-stimulated Colon-26 cells were determined using a luciferase assay and EIA, respectively. Experimental colitis was induced in mice by intrarectal administration of trinitrobenzene sulfonic acid (TNBS). Recombinant proteins were then intrarectally administered into TNBS-induced colitic mice, after which disease activity parameters (body weight, colon length, histological score), as well as interleukin (IL)-6 and MIP-2 levels were determined in inflamed tissues. Flagellin-induced inflammatory cytokine production in vitro was significantly downregulated via αvß3-integrin following pretreatment with wild-type MFG-E8 due to inhibition of NF-κB activation. In vivo, intrarectal treatment with wild-type MFG-E8, but not its mutant counterpart, significantly inhibited body weight loss, colon shortening and histological inflammation induced by TNBS administration. Our findings suggest that MFG-E8 has anti-inflammatory effects on flagellin-induced inflamed intestinal epithelial cells and may be a useful therapeutic agent for colitis.
Subject(s)
Anti-Infective Agents/administration & dosage , Antigens, Surface/administration & dosage , Colitis/drug therapy , Milk Proteins/administration & dosage , Administration, Rectal , Animals , Antigens, Surface/genetics , Cell Line, Tumor , Colitis/chemically induced , Colitis/genetics , Colon/metabolism , Colon/pathology , Cytokines/biosynthesis , Disease Models, Animal , Inflammation/drug therapy , Inflammation/metabolism , Inflammation/pathology , Integrin alpha Chains/genetics , Integrin alpha Chains/metabolism , Integrin beta Chains/genetics , Integrin beta Chains/metabolism , Male , Mice , Mice, Inbred BALB C , Milk Proteins/genetics , NF-kappa B/metabolismABSTRACT
OBJECTIVE: Heparin-binding epidermal growth factor-like growth factor (HB-EGF) has been shown to stimulate the growth and migration of human keratinocytes in an autocrine or paracrine manner. Bearing in mind the preceding narratives, present study was designed to explore the role of HB-EGF on esophageal epithelial cell growth, migration and anti-apoptosis. MATERIAL AND METHODS: HET-1A and TTn cells were treated with recombinant HB-EGF, and cell proliferation and migration were assessed by MTT and Boyden chamber assays, respectively. Anti-apoptotic effects of HB-EGF was studied by Bcl-2/Bcl-xL gene expression and utilizing a TNF-related death apoptosis inducing ligand (TRAIL). RESULTS: Recombinant HB-EGF promotes human esophageal epithelial cell proliferation in a dose dependent manner, where 1 and 10 ng/ml doses were found to be most effective. HB-EGF induced cell migration was noted in TTn, but not in HET-1A cells. Recombinant HB-EGF induced the Bcl-2, Bcl-xL mRNA/protein expression in HET-1A and TTn cells. TRAIL induced the apoptosis in TTn, whereas it was significantly inhibited in HB-EGF treated conditions. Finally, we also revealed HB-EGF induced phosphorylation of EGFR and p38 MAPK in those cell lines, while all cellular functions were repressed by EGFR inhibitor AG1478. CONCLUSION: HB-EGF promotes esophageal epithelial cell proliferation, migration and induces anti-apoptotic gene expression via EGFR/p38 MAPK phosphorylation.
Subject(s)
Apoptosis/drug effects , Cell Proliferation/drug effects , Epithelial Cells/drug effects , Esophagus/growth & development , Intercellular Signaling Peptides and Proteins/pharmacology , TNF-Related Apoptosis-Inducing Ligand/pharmacology , Cells, Cultured , Epithelial Cells/cytology , Epithelial Cells/metabolism , Esophagus/cytology , Esophagus/drug effects , Gene Expression Regulation , Genes, bcl-2/genetics , Heparin-binding EGF-like Growth Factor , Humans , RNA, Messenger/genetics , Receptors, Cell Surface , Recombinant Proteins , Signal Transduction , bcl-X Protein/biosynthesis , bcl-X Protein/geneticsABSTRACT
A unique subset of B cells expressing interleukin-10 (IL-10) and transforming growth factor-ß (TGF-ß) plays an essential role in preventing inflammation and autoimmunity. We investigated the presence of this cell subset in intestines and its role in the pathogenesis of ileitis using SAMP1/Yit and age-matched control AKR/J mice. Mononuclear cells were isolated from mesenteric lymph nodes (MLNs) and the expressions of B220, CD1d, CD5, Toll-like receptor 4 (TLR4) and TLR9 in isolated cells were analysed. Purified B cells were stimulated with lipopolysaccharide (LPS) or CpG-DNA, then IL-10 and TGF-ß(1) expressions were examined by enzyme immunoassay and flow cytometry. Production of IL-1ß by TLR-mediated macrophages co-cultured with or without purified MLN B cells from SAMP1/Yit and AKR/J mice was evaluated. In addition, interferon-γ (IFN-γ) production in intestinal T cells co-cultured with MLN B cells were also assessed in SAMP1/Yit and AKR/J strains. The production levels of IL-10 and TGF-ß(1) stimulated by LPS and CpG-DNA were significantly lower in B cells separated from MLNs from the SAMP1/Yit strain. B cells expressing IL-10 and TGF-ß(1) were mainly located in a population characterized by the cell surface marker CD1d(+) . Interleukin-1ß production by TLR-activated macrophages co-cultured with MLN B cells from SAMP1/Yit mice was significantly higher than that of those from AKR/J mice. Interestingly, IFN-γ production by T cells was noted only when they were co-cultured with SAMP1/Yit but not the AKR/J B cells. These results are the first to show that disorders of regulatory B-cell function under innate immune activation may cause disease pathogenesis in a murine model of Crohn's disease.
Subject(s)
B-Lymphocytes/metabolism , Gene Expression Regulation , Ileum/metabolism , Interleukin-10/biosynthesis , Toll-Like Receptor 4/metabolism , Toll-Like Receptor 9/metabolism , Transforming Growth Factor beta1/biosynthesis , Adjuvants, Immunologic/pharmacology , Animals , Antigens, Differentiation/biosynthesis , Antigens, Differentiation/immunology , B-Lymphocytes/immunology , Crohn Disease/immunology , Crohn Disease/metabolism , Disease Models, Animal , Ileitis/immunology , Ileitis/metabolism , Ileum/immunology , Interferon-gamma/biosynthesis , Interferon-gamma/immunology , Interleukin-10/immunology , Interleukin-1beta/biosynthesis , Interleukin-1beta/immunology , Lipopolysaccharides/pharmacology , Lymphocyte Activation/drug effects , Lymphocyte Activation/immunology , Macrophage Activation/drug effects , Macrophage Activation/immunology , Macrophages/immunology , Macrophages/metabolism , Mesentery/immunology , Mesentery/metabolism , Mice , Mice, Inbred BALB C , Oligodeoxyribonucleotides/pharmacology , Species Specificity , T-Lymphocytes/immunology , T-Lymphocytes/metabolism , Toll-Like Receptor 4/immunology , Toll-Like Receptor 9/immunology , Transforming Growth Factor beta1/immunologyABSTRACT
BACKGROUND/AIMS: Newly developed autofluorescence (AF) imaging (AFI) endoscopy can detect AF emitted by the gastrointestinal wall and may reliably detect tumors or inflammation that block AF. However, the efficacy of AFI endoscopy has not been evaluated for diagnosing the depth of tumor invasion in gastric cancer. METHODS: AF endoscopic images were split into three bands (R, G and B) and expressed as grayscale values. AF indices, defined as the ratio of the G band image grayscale value to that of the R band, were calculated preoperatively. Correlations of AF indices with invasion depth and tumor thickness were assessed. AF indices were calculated preoperatively for 72 gastric cancer lesions without ulceration in 67 patients. The invasion grade of the lesions was classified histologically into 5 groups: M, SM, MP, SS and SE. RESULTS: The mean tumor AF indices for each depth stage were 0.99, 0.77, 0.75, 0.74 and 0.61, respectively. A statistically significant difference was found between group M and the other groups. CONCLUSION: AFI endoscopy may reliably determine the depth of gastric cancer tumor invasion, although an expanded study comprised of larger numbers of subjects and different types of cancers may be required to clearly demonstrate its validity.
Subject(s)
Endoscopy, Gastrointestinal/methods , Neoplasm Invasiveness/pathology , Stomach Neoplasms/pathology , Aged , Aged, 80 and over , Animals , Female , Fluorescence , Humans , Male , Middle Aged , Pilot Projects , ROC Curve , Statistics, Nonparametric , SwineABSTRACT
OBJECTIVE: CD5(+) B cells comprise a unique subset of B cells that modulates innate as well as autoimmune systems. The aim of this study was to investigate alterations of the circulating CD5(+) B-cell subset in patients with inflammatory bowel disease (IBD) by evaluating various clinical parameters, including therapeutic regimens. MATERIAL AND METHODS: Thirty-four patients with ulcerative colitis (UC), 19 patients with Crohn's disease (CD), and 46 healthy control subjects were enrolled in this study. CD5(+) B cells in peripheral blood collected from each subject were analyzed by flow cytometry. Multiple regression analysis was carried out to evaluate the factors related to the circulating CD5(+) B-cell subset in the IBD patients. In an in vitro examination, dexamethasone-induced apoptosis in peripheral blood B cells was examined by detecting cell surface binding of the annexin-V antibody. RESULTS: Age and gender in the control subjects did not influence the circulating CD5(+) B-cell subset. Multiple regression analysis showed that the presence of UC, corticosteroid therapy, and number of white blood cells in peripheral blood each had a significant influence in decreasing the number of circulating CD5(+) B cells in the IBD patients. Furthermore, in vitro results showed that dexamethasone treatment significantly induced apoptosis in CD5(+) B cells, though apoptosis was similarly observed in CD5(-) B cells. CONCLUSIONS: CD5(+) B cells may be involved in the pathogenesis of UC, and modulation of this subset by corticosteroid therapy may play a role in the treatment of IBD patients.
Subject(s)
B-Lymphocyte Subsets/metabolism , B-Lymphocytes/metabolism , CD5 Antigens/blood , Inflammatory Bowel Diseases/blood , Inflammatory Bowel Diseases/immunology , Adolescent , Adult , Aged , CD5 Antigens/immunology , Case-Control Studies , Female , Glucocorticoids/therapeutic use , Humans , Inflammatory Bowel Diseases/drug therapy , Male , Middle AgedABSTRACT
The lactogenic hormone prolactin (PRL) regulates milk protein gene expression in mammary glands. To maintain homeostatic balance in the body, milk fat globule epidermal growth factor 8 (MFG-E8) is vital for phagocytic clearance of apoptotic cells. We investigated the effects of PRL on MFG-E8 expression in macrophages by evaluating its promoter function. Macrophages were stimulated with PRL, and the expression of MFG-E8 was determined using real-time PCR and Western blotting. The role of MFG-E8 on phagocytosis of apoptotic cells in PRL-treated macrophages was assessed using microscopy, while the response of PRL to MFG-E8 expression was evaluated using luciferase assay. Following treatment with PRL, significant up-regulations of the PRL receptor and MFG-E8 were observed in macrophages, though PRL-treated macrophages more efficiently engulfed apoptotic cells. The results of MFG-E8 promoter analysis showed considerable up-regulation of promoter activity in macrophages following PRL treatment and results from mutation analysis of the MFG-E8 promoter suggested that the C/EBPbeta binding site was responsible for PRL-induced activation of the MFG-E8 promoter. C/EBPbeta activity was found to be up-regulated in PRL-treated cells as revealed by an electrophoretic mobility shift assay (EMSA). In conclusion, PRL is a potent inducer of MFG-E8 expression in macrophages, while its effect is mediated by the presence of a responsive element in the MFG-E8 promoter.