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Pedica F Ligorio C Tonelli P Bartolini S Baccarini P 《Virchows Archiv : an international journal of pathology》2008,452(1):57-63
Crohn’s disease (CD) is a chronic inflammatory bowel disorder of unknown etiology. An involvement of the intestinal lymphatic
system has been suggested. Recently, monoclonal antibodies have become available to distinguish lymphatic vessels from blood
vessels. The aim of the study was to examine the distribution of lymphatic vessels in ileal and colic walls of patients affected
by CD and compare it with healthy controls and other inflammatory bowel diseases. Twenty-eight cases of CD, 13 cases of other
inflammatory bowel diseases, and 10 normal ileal and colic walls were studied. Immunohistochemical staining was performed
using the monoclonal antibody D2-40. Quantification of lymphatic vessels was performed by identifying four fields with high
density of lymphatics and then counting the number of lymphatic vessels at high resolution. Lymphatic diameter was also evaluated
by using an ocular micrometer. Lymphatic vessels showed the highest density in CD specimens. The median number of lymphatics
was significantly higher both in ileal and colic samples of CD than the other inflammatory diseases as well as normal controls.
Moreover, in patients with CD, diffuse lymphangiectasia was also observed. The present data suggest that lymphangiogenesis
and lymphangiectasia probably play a role in the pathogenesis of CD. 相似文献
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《Expert Review of Clinical Immunology》2013,9(6):831-841
The cryopyrin-associated periodic syndromes (CAPS) are a group of rare inherited inflammatory diseases associated with overproduction of IL-1β. Canakinumab, developed by Novartis AG (Basel, Switzerland), is an intravenously or subcutaneously administered, fully human monoclonal antibody that neutralizes the bioactivity of human IL-1β. Canakinumab has promising clinical safety and pharmacokinetic properties, and has demonstrated potential for the treatment of CAPS. Canakinumab was recently granted EU orphan drug status for systemic-onset juvenile idiopathic arthritis, and early clinical trials have established that administration of canakinumab every 2 weeks is both safe and effective. Subcutaneous canakinumab (approved formulation) offers some advantages over the existing IL-1β-blocking treatment, anakinra, which must be injected daily and is often not well tolerated by patients. The long-term safety of all targeted anti-IL-1 therapies in CAPS remains an unanswered question owing to the relatively short clinical experience with these agents; as canakinumab produces sustained IL-1 suppression, vigilance is necessary to diagnose the development of adverse events, especially any associated infections. 相似文献
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The present microelectrode experiments on fused respiratory epithelial cells of cystic fibrosis (CF) origin and non-CF origin aim at characterizing the molecular basis of the Cl– conductances regulated by cyclic adenosine monophosphate (cAMP) or respectively Ca2+, as described in the preceding publication. Cell membrane potential (V
m) and resistance (R
m) were recorded as well as their response to substitution of 90% of bath Cl– by isethionate (V
m,ISE), by I– (V
m,I), or by other halide anions. Fused CF cells had significantly (P<0.05) higher control V
m values (P–18.0 ±9.4 mV, ±SD, n=68) than fused non-CF cells (–12.5±6.6 mV, n=69) and responded to the Ca2+ ionophore A23187 with an increase in the V
m response to Cl– substitution, but did not respond to forskolin. This indicates that CF cells express only the Ca2+-stimulated Cl– conductance. Injection of the antibody M3A7 against a fusion protein containing amino acids 1195 to 1480 of the CF gene product into young, forskolin-stimulated or old non-CF cells decreased V
m,ISE and V
m,I within 15 min to values observed in CF cells. This indicates inhibition of the cAMP-stimulated Cl– conductance and supports the molecular identity of this conductance with the CF gene product. However, the slow onset of inhibition does not allow secondary effects to be excluded and a slight fall in R
m remains unexplained. Stimulation of the Ca2+-regulated Cl– conductance was not impaired. Injection of M3A7 into CF cells or of a control antibody in non-CF cells had no effect. In the search for the single-channel equivalent of the Ca2+-stimulated Cl– conductance we injected a concentrated placental cytosol fraction containing a cytosolic inhibitor of the outwardly rectifying intermediate conductance (ORIC) Cl– channel into fused non-CF cells stimulated either with A23187 or forskolin. However, no effect was observed. This speaks against a role of the ORIC Cl– channel in the Ca2+-activated Cl– conductance, although it cannot definitely be excluded. 相似文献
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The epithelial cell surface of the endometrium undergoes substantial biochemical changes to allow embryo attachment and implantation in early pregnancy. We hypothesized that tissue macrophages influence these events to promote uterine receptivity. To investigate the role of macrophages in regulating epithelial cell expression of genes linked to glycan-mediated embryo adhesion, Ishikawa, RL95-2 and HEC1A endometrial epithelial cells were cultured alone or with unactivated or lipopolysaccharide-activated monocytic U937 cells, separated using transwell inserts. Expression of mRNAs encoding two α1,2-fucosyltransferases (FUT1, FUT2) was increased in all three epithelial cell lines following co-culture with U937 cells, and was associated with increased fucosylation of cell surface glycoproteins detected using lectins from Ulex europaeus (UEA-1) and Dolichos biflorus (DBA). FUT1 induction by U937 cells also occurred in primary endometrial epithelial cells collected in luteal but not proliferative phase. Activation of the interleukin-6 (IL6)/leukemia inhibitory factor (LIF) cytokine signaling pathway with phosphorylation of STAT3 and elevated SOCS3 mRNA expression was evident in epithelial cells stimulated by U937 co-culture. Several recombinant macrophage-secreted cytokines exerted stimulatory or inhibitory effects on FUT1 and FUT2 mRNA expression, and the macrophage-derived cytokine LIF partially replicated the effects of U937 cells on both FUT1 and FUT2 expression and UEA-1 and DBA lectin reactivity in Ishikawa cells. These results suggest that macrophage-derived factors including LIF might facilitate development of an implantation-receptive endometrium by regulating surface glycan structures in epithelial cells. Abnormal phenotypes or altered abundance of uterine macrophages could contribute to the pathophysiology of primary unexplained infertility in women. 相似文献
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The cryopyrin-associated periodic syndromes (CAPS) are a group of rare inherited inflammatory diseases associated with overproduction of IL-1β. Canakinumab, developed by Novartis AG (Basel, Switzerland), is an intravenously or subcutaneously administered, fully human monoclonal antibody that neutralizes the bioactivity of human IL-1β. Canakinumab has promising clinical safety and pharmacokinetic properties, and has demonstrated potential for the treatment of CAPS. Canakinumab was recently granted EU orphan drug status for systemic-onset juvenile idiopathic arthritis, and early clinical trials have established that administration of canakinumab every 2 weeks is both safe and effective. Subcutaneous canakinumab (approved formulation) offers some advantages over the existing IL-1β-blocking treatment, anakinra, which must be injected daily and is often not well tolerated by patients. The long-term safety of all targeted anti-IL-1 therapies in CAPS remains an unanswered question owing to the relatively short clinical experience with these agents; as canakinumab produces sustained IL-1 suppression, vigilance is necessary to diagnose the development of adverse events, especially any associated infections. 相似文献
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Huang Yunxiu Chen Linmu Zhu Baofang Han Hui Hou Yanfang Wang Weijia 《Clinical and experimental medicine》2021,21(1):73-78
Clinical and Experimental Medicine - The objective of the study was to investigate the value of anti-α-enolase antibody (Ab) combined with RDW in evaluating the activity of systemic lupus... 相似文献
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Zhen-Jun Shao Xing-Ping Li Yi-Ying Wu Xiao-Wei Zheng Juan Huang Wen-Long Deng 《Food and Agricultural Immunology》2010,21(4):361-367
In this study we observed the enhanced mRNA and protein expression of human β-defensin-1 (hBD-1) induced by dehydroandrographolide (DA) in human gland epithelial cells and explored the mechanism of DA on infection treatment. Human pulmonary gland epithelial cells were incubated with DA, then were harvested for hBD-1 expression detection. The mRNA expression of hBD-1 was detected by RT-PCR while the protein expression was detected by Western blot. It was found that DA can up-regulate mRNA and protein expression of hBD-1, the optimal concentration was 80 µM, the maximal expression of hBD-1 mRNA and protein occurred after 8 h. The DA can up-regulate mRNA and protein expression of hBD-1 in human gland epithelial cells. It suggests that β-defensin-1 may play an important role in the treatment of infective diseases with DA. 相似文献
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Kanet Pakdeepak Supansa Pata Sawitree Chiampanichayakul Watchara Kasinrerk 《Journal of immunoassay & immunochemistry》2016,37(6):564-571
Monoclonal antibodies against α-globin containing human Hbs, named AMS-Alpha1 and AMS-Alpha 2, were produced by the hybridoma technique using spleen cells enriched by the newly developed B lymphocyte enrichment protocol. These two monoclonal antibodies were of IgM class, reacting to only intact form of human Hbs A, A2, E, and F, which contain α-globin chain. By the indirect ELISA, the AMS-Alpha1 and AMS-Alpha 2 quantified less amount of α-globin chain containing hemoglobins in HbH disease than the SEA-α thalassemia 1 carriers and normal individuals. It was thus anticipated that these monoclonal antibodies can be used for detecting Hb Bart’s hydrops fetalis in which no α-globin chain is produced. 相似文献
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The ultraviolet (UV)-induced (6–4) pyrimidine–pyrimidone photoproduct [(6–4) PP] confers a large structural distortion in DNA. Here we examine in human cells the roles of translesion synthesis (TLS) DNA polymerases (Pols) in promoting replication through a (6–4) TT photoproduct carried on a duplex plasmid where bidirectional replication initiates from an origin of replication. We show that TLS contributes to a large fraction of lesion bypass and that it is mostly error-free. We find that, whereas Pol η and Pol ι provide alternate pathways for mutagenic TLS, surprisingly, Pol ζ functions independently of these Pols and in a predominantly error-free manner. We verify and extend these observations in mouse cells and conclude that, in human cells, TLS during replication can be markedly error-free even opposite a highly distorting DNA lesion. 相似文献
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《Journal of histotechnology》2013,36(1):3-10
AbstractImmunohistochemistry (IHC) is a method of routine use in histopathology for the diagnosis and differentiated diagnosis of liver diseases. IHC staining in human hepatic tissue is often obstructed by the presence of non-specific antibody interactions, the cause of high background staining. Cognizant of previous encounters with high levels of background in human hepatic tissue when using polyclonal antibodies, the potential causes were systemattically investigated by testing the impact of different antibody variants and assay conditions. Firstly, IHC was performed on hepatic tissue with immunized and non-immunized polyclonal antibodies and it was demonstrated that immunized antibodies pervaded the samples with non-specific staining, while non-immunized antibodies produced insignificant background. Following, the primary antibody was co-incubated with different concentrations of protein blocks, acid/base solutions, crystalline ionic compounds, and combinations of these individual components in order to assess their potency in removing background staining. Optimization of these assay conditions revealed that background could be significantly reduced by the addition of sodium chloride to primary antibody solution and/or protein block to sodium azide-stabilized Tris-HCl buffer, while little beneficial effect was observed from co-incubating the primary antibody with non-serum protein block/goat serum, hydrochloric acid/sodium hydroxide, or a higher concentration of peroxidase block. Together, the results suggest that hepatic background in IHC is mainly attributed to strong ionic interactions between the light chains of polyclonal antibodies and polar molecules specifically prevalent in hepatic tissue, which can be reduced by increasing the ionic strength of the antibody–antigen reaction microenvironment. Thus, the findings can enhance the specificity of hepatic tissue immunoassays and improve IHC studies in human liver tissues using polyclonal antibodies. 相似文献
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Fabio Caradonna Ilenia Cruciata Ilaria Schifano Chiara La Rosa Flores Naselli Roberto Chiarelli Anna Perrone Carla Gentile 《Inflammation research》2018,67(4):327-337
Objective and design
Epigenetic regulation is important in the activation of inflammatory cells. In the present study, we evaluated if DNA-methylation variations are involved in Interleukin-1β (IL-1β)-induced intestinal epithelial cells activation.Materials and methods
Differentiated Caco-2 cells were exposed to IL-1β or to 5-azadeoxycytidine (5-azadC) for 24 or 48 h. Genome-wide methylation status was evaluated, while DNA methylation status at the promoter region of the gene encoding interleukin-6, 8 and 10 (IL-6, 8 and 10) was estimated. The levels of the corresponding gene products as well as DNA methyltransferases (DNMTs) quantity were assessed.Results
IL-1β decreased genomic methylation of human intestinal epithelial cells and induced demethylation at cg-specific sites at the promoter of pro-inflammatory genes IL6 and IL8; conversely it did not change the methylation of the IL10 promoter. IL-1β also increased the release of IL-6 and IL-8 but did not change the IL-10 expression. Finally, cell exposure to IL-1β decreased the DNMT3b expression, increased DNMT3a and was not able to change DNMT1 expression.Conclusions
Our results suggest a potential role of IL-1β as modulator of DNA methylation in activated differentiated Caco-2 cell line.16.
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Omagari D Takenouchi-Ohkubo N Endo S Ishigami T Sawada A Moro I Asano M Komiyama K 《Clinical and experimental immunology》2011,165(1):85-93
Intestinal epithelial cells (IECs) play an important role in protecting the intestinal surface from invading pathogens by producing effector molecules. IECs are one of the major sources of human beta-defensin 2 (hBD-2), and can produce it in response to a variety of stimuli. Although IECs express Toll-like receptor 3 (TLR-3) and can respond to its ligand, double-stranded RNA (dsRNA), hBD-2 expression in response to dsRNA has not been elucidated. In the present study, using an artificial analogue of dsRNA, polyinosinic-polycytidylic acid (poly I:C), we investigated whether the human IEC line, HT-29, can produce hBD-2 in response to poly I:C. HT-29 cells can express hBD-2 mRNA only when stimulated with poly I:C. The induction of hBD-2 mRNA expression was observed at 3 h after stimulation and peaked at 12 h of post-stimulation. Pre-incubation of the cells with nuclear factor kappa B (NF-κB)-specific inhibitor, l-1-4'-tosylamino-phenylethyl-chloromethyl ketone (TPCK) and isohelenine abolished the expression of hBD-2. Detection of the poly I:C signal by TLR-3 on the surface of HT-29 cells was revealed by pre-incubating the cells with anti-TLR-3 antibody. The 5'-regulatory region of the hBD-2 gene contains two NF-κB binding sites. A luciferase assay revealed the importance of the proximal NF-κB binding site for poly I:C-induced expression of hBD-2. Among NF-κB subunits, p65 and p50 were activated by poly I:C stimulation and accumulated in the nucleus. Activation of the p65 subunit was investigated further by determining its phosphorylation status, which revealed that poly I:C stimulation resulted in prolonged phosphorylation of p65. These results indicate clearly that NF-κB plays an indispensable role in poly I:C induced hBD-2 expression in HT-29 cells. 相似文献
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Ohashi T Iizuka S Shimada Y Eto Y Ida H Hachimura S Kobayashi H 《Molecular genetics and metabolism》2011,103(1):98-100
Animal and human studies of enzyme replacement therapy for Pompe disease have indicated that antibodies generated against the infused recombinant human acid α-glucosidase (rhGAA) can negatively impact therapeutic outcome. In this study, we show that oral administration of rhGAA into mice can reduce the titer of anti-rhGAA antibody following immunization with rhGAA. Oral administration of rhGAA is safe and antigen specific, it offers advantages over other immunosuppressive drugs. 相似文献
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Meng Ling Moi Chang-Kweng Lim Shigeru Tajima Akira Kotaki Masayuki Saijo Tomohiko Takasaki Ichiro Kurane 《Journal of clinical virology》2011,52(3):225-230