transforming-growth-factor-beta and Peritonitis

Topics: Animals; Carrier Proteins; Depression, Chemical; Diabetic Nephropathies; Dialysis Solutions; Enzyme Activation; Extracellular Matrix Proteins; Fibrosis; Gene Expression Regulation; Glomerular Mesangium; Glucose; Glycation End Products, Advanced; Glycosaminoglycans; Humans; Intracellular Signaling Peptides and Proteins; Isoenzymes; Latent TGF-beta Binding Proteins; NF-kappa B; Peritoneal Dialysis; Peritoneal Dialysis, Continuous Ambulatory; Peritoneum; Peritonitis; Protein Kinase C; RNA, Messenger; Tetradecanoylphorbol Acetate; Time Factors; Transforming Growth Factor beta

Peritoneal dialysis (PD) is essential for patients with end-stage renal disease. Peritoneal fibrosis (PF) is a complex inflammatory, fibrogenic process. No effective treatments are available to prevent these processes. Hepatocyte growth factor (HGF) possesses anti-inflammatory and anti-fibrotic properties. The aim of this study was to analyze whether HGF suppresses MGO-induced peritoneal inflammation and fibrosis in a mouse model.. PF was induced by intraperitoneal (IP) injections of MGO for 14 days. C57/BL/6 mice were divided into three groups: Sham group (only vehicle); Sham + MGO group (PF induced by MGO); and HGF + MGO group (PF mice treated with recombinant human-HGF). PF was assessed from tissue samples by Masson's trichrome staining. Inflammation and fibrosis-associated factors were assessed by immunohistochemistry and quantitative real-time PCR.. MGO-injected mice showed significant thickening of the submesothelial compact zone with PF. Treatment with HGF significantly reduced PM thickness and suppressed the expression of collagen I and III and α-SMA. Expression of profibrotic and proinflammatory cytokines (TGF-β, TNF-α, IL-1β) was reduced by HGF treatment. The number of macrophages, and M1 and M2 macrophage-related markers, such as CD86, CD206, and CD163, was reduced in HGF + MGO mice.. HGF attenuates MGO-induced PF in mice. Furthermore, HGF treatment reduces myofibroblast and macrophage infiltration, and attenuates the upregulated expression of proinflammatory and profibrotic genes in peritoneal tissues. HGF might be an effective approach to prevent the development of PF in patients undergoing PD.

Topics: Actins; Animals; Collagen Type I; Collagen Type III; Disease Models, Animal; Gene Expression; Hepatocyte Growth Factor; Interleukin-1beta; Macrophages; Male; Matrix Metalloproteinase 3; Matrix Metalloproteinase 9; Mice; Mice, Inbred C57BL; Myofibroblasts; Peritoneal Fibrosis; Peritonitis; Pyruvaldehyde; Recombinant Proteins; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Up-Regulation

Regulatory T cells (Tregs) and transforming growth factor β (TGF-β) are believed to play key roles in both postoperative pro-inflammatory and anti-inflammatory responses of malignancies. Recombinant human thrombomodulin (rTM) is implied to inhibit the interaction between TGF-β and Tregs. The aim of this study is to evaluate the antitumor effects of rTM against gastrointestinal tumors under systemic inflammation. Mice were subjected to cecal ligation and puncture and percutaneous allogeneic tumor implantation. rTM were introduced by percutaneous injection into the abdominal cavity. The effects of rTM were evaluated by weight of implanted tumor, proportion of Tregs in peripheral blood lymphocytes (PBL) and tumor infiltrating lymphocytes (TIL) and temporal evaluation of serum cytokines. The effect of rTM was also evaluated on the in vitro differentiation of naïve T cells into induced Tregs induced by TGF-β and interleukin (IL) -2. rTM significantly inhibited the proliferation of the implanted tumor cells in an inflammation-dependent manner. rTM also reduced the fractions of regulatory T cells and induced regulatory T cells among both PBL and TIL. Temporal evaluation of serum cytokine levels in the model mice showed that rTM significantly suppressed the increases in the serum levels of IL-2 and TGF-β. An in vitro differentiation assay revealed that rTM inhibited the differentiation of naïve T cells into Tregs triggered by IL-2- and TGF-β. rTM has suppressive effects on inflammation-induced gastrointestinal tumor growth by suggestively affecting differentiation of Tregs.

Topics: Animals; Cell Differentiation; Cell Line, Tumor; Disease Models, Animal; Female; Gastrointestinal Neoplasms; Inflammation; Mice; Mice, Inbred BALB C; Peritonitis; Recombinant Proteins; T-Lymphocytes, Regulatory; Thrombomodulin; Transforming Growth Factor beta

To evaluate the effects of platelet rich plasma (PRP) on the healing of fascia wherein peritonitis has been created.. Twenty eight Wistar Albino rats were divided into four groups. Only a primary fascial repair following laparotomy was performed on Group 1, a primary fascial repair performed and PRP treatment applied following laparotomy on Group 2, and a fecal peritonitis created following laparotomy and a primary fascial repair carried out on Group 3. A fecal peritonitis was created following laparotomy and primary fascial repair and PRP treatment on the fascia was carried out on Group 4.. TNF-α was found to be significantly lower in the control group (Group 1). It was detected at the highest level in the group in which fecal peritonitis was created and PRP applied (Group 4). TGF-β was determined as being significantly higher only in Group 4. Histopathologically, the differences between the groups in terms of cell infiltration and collagen deposition were not found to be significant.. When platelet rich plasma was given histologically and biochemicaly as wound healing parameters cellular infiltration, collagen accumulation, and tissue hydroxyiproline levels were not increased but neovascularization, fibroblast activation and TNF Alfa levels were increased and PRP accelerated wound healing.

Topics: Animals; Collagen; Endopeptidases; Fascia; Gelatinases; Hydroxyproline; Membrane Proteins; Models, Animal; Neovascularization, Physiologic; Peritonitis; Platelet-Rich Plasma; Random Allocation; Rats, Wistar; Serine Endopeptidases; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Wound Healing

Peritonitis, the most important limitation of peritoneal dialysis (PD), could be detected by biomarkers in dialysate effluent, representing a noninvasive method to indirectly assess the peritoneum status. The aim of our study was to test high mobility group box 1 (HMGB1) in PD patients, evaluating its role as precocious marker of peritoneum damage during peritonitis. Transforming growth factor (TGF)-β was correlated with peritoneal transport characteristics.. Six patients, treated by ambulatory PD, were enrolled. Samples were collected at the onset of peritonitis (T1) and every day until its resolution (T-end). Serum (s) and peritoneal (p) white blood cell (WBC) count was also evaluated. Peritoneal Equilibration Test evaluated the filter activity of peritoneum.. In patients with acute peritonitis, the highest serum and peritoneal HMGB1 values (64 ± 3.6 and 70 ± 5.3 ng/mL, respectively) were assessed, with a progressive decrease of their levels at the resolution time (T-end: sHMGB1:36 ± 2.5; pHMGB1:30.5 ± 7.0 ng/mL). While no differences of sWBC and pWBC were observed between baseline and T-end values, pHMGB1 levels remained higher at T-end than those observed at T0 (pHMGB1:30.5 ± 7.0 versus 6.9 ± 3.6; p < 0.0001). TGF-β levels were higher in patients with low peritoneal permeability than in medium or high transporter patients (81 ± 15.5 versus 24.3 ± 7.5 pg/mL; p = 0.01). An inverse correlation was found between TGF-β levels and dialysate/plasmatic creatinine values (r = -0.83; p = 0.03).. HMGB1 represents a useful biomarker for peritoneum evaluation in PD patients. A prognostic role of this alarmin, as a marker of response to therapy, could be hypothesized. TGF-β could predict the peritoneal transport status and dialysis technique adequacy.

Topics: Adolescent; Biomarkers; Child; Child, Preschool; Disease Progression; Female; HMGB1 Protein; Humans; Kidney Failure, Chronic; Male; Peritoneal Dialysis; Peritonitis; Prognosis; Transforming Growth Factor beta

Duodenal injuries, though rare, carry high rates of morbidity and mortality. The aim of the study was to evaluate the healing of the duodenal wall with the use of a Small Intestinal Submucosa (SIS) patch.. We studied 40 Wistar-Albino rats divided into two groups. In group A, we created a small defect in the duodenal wall, which was immediately covered with a SIS patch. In group B, the SIS patch was sutured over the defect after 6-8 hours, in order to induce peritonitis. The animals of both groups were sacrificed after 2, 6, 12 and 16 weeks respectively. In addition, we studied the immunohistochemical expression of TGF-β, which is a major constituent of SIS, and plays a central role in the healing process.. Histology showed progressive development of the layers of the duodenal wall over the patch as early as the 2nd week in some of the animals of group A. Mucosa developed later on in the animals of group B, presumably due to the more intense inflammation elicited by peritonitis. Expression of TGF-β was initially more pronounced in the epithelial cells of the regenerating mucosa of animals of group A, but it was maintained in higher levels in the animals of group B, which showed delayed mucosa degeneration.. SIS appears to be both efficient and safe for the management of duodenal trauma. TGF-β seems to play an important role in the healing process, inducing regeneration of the stroma, and controlling epithelial growth.

Topics: Animals; Duodenum; Intestinal Mucosa; Peritonitis; Rats; Rats, Wistar; Time Factors; Transforming Growth Factor beta; Wound Healing; Wounds, Penetrating

It has been shown recently that significant number (to 40% from total population) of macrophage foam cells (MFC) is formed during early time (24 h) of zymosan-induced peritonitis resolution and agonists of peroxisome proliferation activated receptors-α, -γ (PPAR-α, -γ) exert anti-inflammatory action, protecting their formation (Dushkin et al., 2007). The work is devoted to investigate of the influence of cholesterol-containing liposomes (CHL) on dinamic of zimozan-induced peritonitis in C57Bl/6 mice. The accumulation of cholesterol, the change of cytokine production, PPAR-γ activity and cholesterol efflux in macrophages of C57Bl/6 mice has been investigated. The infiltration of neutrophils, amounts of mononuclear cells and MFC formation were significantly increased in peritonel cavity of zymosan-induced mice that led to in expansion of the period of inflammatory resolution and of the period of MFC resolution. If macrophages obtained after zymosan injection mainly accumulated triglycerides (TG) and at high speed incorporated [1-14C]oleate into TG, the injection of CHL after zymosan-indused inflammation lead to dramatic promotion MFC containing primarily free cholesterol and Ch ethers and been aggravation of [1-14C]oleate incorporation into cholesterol ethers in macrophages (mainly for 2 days). It has to shown that CHL against a background of inflammation promoted reduction of fluorescent NBD-cholesterol efflux from macrophages throughout the studied period (5 days) whereas zymosan inhibited cholesterol efflux at the early stages of inflammation (1 and 2 days), then, on 3ed day, the cholesterol efflux was recovered and increased on day 5. At the same time CHL stimulated the production of TNFα and TGFβ and inhibited the production of IL-10 and DNA-binding activity of PPAR-γ macrophages obtained at early as well as late stages of zymosan-induced peritonitis (compared with injection zymosan only). Thus, accumulation of cholesterol in inflammatory macrophages and promotion of MFC formation prolog timely resoluti- on of acute inflammation inducing alteration of pro- and anti-inflammatory cytokine balance and evoking the repression of macrophage DNA-binding activity of PPAR-γ and cholesterol efflux.

Topics: 4-Chloro-7-nitrobenzofurazan; Animals; Biological Transport; Cholesterol; Foam Cells; Inflammation; Interleukin-10; Liposomes; Male; Mice; Mice, Inbred C57BL; Neutrophil Infiltration; Neutrophils; Oleic Acid; Peritonitis; PPAR gamma; Transforming Growth Factor beta; Triglycerides; Tumor Necrosis Factor-alpha; Zymosan

Dietary n-3 polyunsaturated fatty acids (PUFA) influence the inductive phase of inflammation but less is known about their effects on the resolution phase. This study examined the effects of dietary fish oil on induction and resolution of antigen-induced inflammation in mice. Mice were fed a control diet with or without 2.8% fish oil, immunized twice with methylated BSA (mBSA) and inflammation induced by intraperitoneal injection of mBSA. Prior to and at different time points after mBSA administration, peritoneal cells were analyzed and expression of surface molecules determined by flow cytometry. Concentration of chemokines, cytokines and soluble cytokine receptors was determined by ELISA. Mice fed the fish oil diet had fewer peritoneal neutrophils, shorter resolution interval and lower levels of pro-inflammatory cytokines and chemokines than mice fed the control diet. In mice fed the fish oil diet there was an early peak in peritoneal levels of the immunosuppressive molecules sIL-6R and TGF-β, that was not seen in mice fed the control diet. In the resolution phase, peritoneal macrophages from mice fed the fish oil diet expressed more of the atypical chemokine receptor D6 and peritoneal TGF-β levels were higher than that in mice fed the control diet. Furthermore, in the late-resolution phase there were more peritoneal eosinophils and macrophages in mice fed the fish oil diet than in mice fed the control diet. These results demonstrate a suppressive effect of n-3 PUFA on the inductive phase of inflammation and indicate an enhancing effect of n-3 PUFA on resolution of inflammation.

Topics: Animals; Chemokine CCL11; Chemokine CXCL1; Dietary Fats, Unsaturated; Fatty Acids, Omega-3; Female; Fish Oils; Granulocyte Colony-Stimulating Factor; Inflammation; Interleukin-6; Macrophages, Peritoneal; Mice; Mice, Inbred C57BL; Peritonitis; Receptors, Interleukin-6; Serum Albumin, Bovine; Transforming Growth Factor beta

To facilitate the targeting to inflammation sites of small anti-inflammatory peptides, with short half-lives, by fusion with the latency-associated peptide (LAP) of transforming growth factor β1 through a cleavable matrix metalloproteinase (MMP) linker. This design improves efficacy, overcoming the limitations to their clinical use.. We generated latent forms of vasoactive intestinal peptide (VIP), α-melanocyte-stimulating hormone (MSH) and γ(3)MSH by fusion to LAP through an MMP cleavage site using recombinant DNA technology. The biological activities of these latent therapeutics were studied in vivo using monosodium urate (MSU)-induced peritonitis and collagen-induced arthritis (CIA) models. We assessed gene therapy and purified protein therapy.. The recruitment of the polymorphonuclear cells induced by MSU injection into mouse peritoneal cavity was reduced by 35% with γ(3)MSH (1 nmol), whereas administration of a much lower dose of purified latent LAP-MMP-γ(3)MSH (0.03 nmol) attenuated leucocyte influx by 50%. Intramuscular gene delivery of plasmids coding LAP-MMP-VIP and LAP-MMP-αMSH at disease onset reduced the development of CIA compared with LAP-MMP, which does not contain any therapeutic moiety. Histological analysis confirmed a significantly lower degree of inflammation, bone and cartilage erosion in groups treated with LAP-MMP-VIP or LAP-MMP-αMSH. Antibody titres to collagen type II and inflammatory cytokine production were also reduced in these two groups.. Incorporation of small anti-inflammatory peptides within the LAP shell and delivered as recombinant protein or through gene therapy can control inflammatory and arthritic disease. This platform delivery can be developed to control human arthritides and other autoimmune diseases.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Experimental; Cytokines; Drug Delivery Systems; Drug Design; Drug Evaluation, Preclinical; Genetic Therapy; Half-Life; Male; Melanocyte-Stimulating Hormones; Mice; Mice, Inbred DBA; Peptide Fragments; Peritonitis; Recombinant Fusion Proteins; Tissue Distribution; Transforming Growth Factor beta; Treatment Outcome; Vasoactive Intestinal Peptide

Peritonitis is a common and severe complication of peritoneal dialysis (PD). Although TGF-beta is a key mediator in peritoneal fibrosis with chronic PD, its role in acute peritoneal inflammation remains unclear.. Potential role of TGF-beta signalling in acute peritonitis was investigated in a rat model by infecting peritoneum with E. coli and in primary culture of peritoneal mesothelial cells (PMC) by LPS.. We found that a single infection of E. coli caused an acute, but transient peritonitis by a significant increase in ascites white blood cells (WBC), peritoneal CD45+ leukocytes, upregulation of TNFalpha, activation of NF-kappaB/p65 and impaired peritoneal function (all P < 0.01). Interestingly, spontaneous recovery of acute peritonitis occurred with upregulation of TGF-beta1 and activation of Smad2/3, suggesting a protective role of TGF-beta signalling in acute peritonitis. This was demonstrated by the finding that blockade of the TGF-beta signalling pathway with gene transfer of Smad7 inactivated peritoneal Smad2/3 but worsened E. coli-induced, NF-kappaB-dependent peritoneal inflammation and peritoneal dysfunction (all P < 0.01). Furthermore, studies in vitro also found that impaired TGF-beta signalling by overexpressing Smad7 in PMC were able to overcome the inhibitory effect of TGF-beta on LPS-induced, NF-kappaB-mediated peritoneal inflammation.. Results from this study demonstrate that TGF-beta signalling is essential in protection against acute peritoneal inflammation induced by bacterial infection.

Topics: Animals; Escherichia coli Infections; Male; Peritonitis; Rats; Rats, Sprague-Dawley; Signal Transduction; Smad7 Protein; Transforming Growth Factor beta

The association between peritoneal solute transport rates (PSTRs) and inflammatory markers in patients on peritoneal dialysis (PD) is still under investigation. We aimed to elucidate their relationship during the first year on PD.. We performed a prospective observational study with 187 incident PD patients who were treated with either biocompatible solution (BCS) or conventional solution (CS). Peritoneal dialysate effluent (PDE) and blood samples for the markers and the calculation of mass transfer area coefficient of creatinine (MTAC) were performed at 1, 6 and 12 months after commencing PD.. Of the 187 enrolled patients, 110 completed a 1-year study protocol. All PDE markers [interleukin-6 (IL-6), transforming growth factor-beta (TGF-beta), TGF-beta-induced gene-h3 (beta ig-h3), vascular endothelial growth factor (VEGF)] except CA125 increased over time, whereas PSTRs, high-sensitivity C-reactive protein (hs-CRP) and serum IL-6 levels did not change. Serum albumin and log PDE appearance rates (ARs) of IL-6, TGF-beta and CA125 predicted MTAC. The Delta value (12-month minus 1-month) of PDE AR of IL-6 was correlated with those of all other PDE markers. Both 12-month IL-6 and Delta IL-6 ARs in PDE were highest in the upper Delta MTAC tertile. PSTRs in the CS group, unlike BCS, had a tendency to increase over time, demonstrating a time-by-group interaction. Solution type and MTAC were not associated with patient and technique survival.. The change in PSTR during the first year of PD is related to PDE IL-6 AR, which may represent intraperitoneal inflammation; however, there does not seem to be a close association between PSTR and the degree of systemic inflammation.

Topics: Adult; Aged; Biological Transport, Active; Biomarkers; CA-125 Antigen; Creatinine; Cytokines; Dialysis Solutions; Female; Humans; Inflammation; Inflammation Mediators; Interleukin-6; Kaplan-Meier Estimate; Kidney Failure, Chronic; Male; Membrane Proteins; Middle Aged; Peritoneal Dialysis; Peritoneum; Peritonitis; Permeability; Prospective Studies; Transforming Growth Factor beta

We measured the neutrophil gelatinase-associated lipocalin (NGAL) concentration in peritoneal dialysate effluent (PDE) collected following an acute episode of continuous ambulatory peritoneal dialysis (CAPD)-related peritonitis.. NGAL concentration in PDE increased in the first 3 days after developing peritonitis and correlated well with the neutrophil count. In patients with culture-negative peritonitis, the NGAL in PDE was lower than that in patients with gram-positive or gram-negative peritonitis. Apart from providing additional diagnostic support to bacterial-induced peritonitis, measurement of NGAL in PDE may be useful to differentiate the neutrophil-dependent culture-negative peritonitis from that associated with non-bacterial or non-cellular etiologies.. Human peritoneal mesothelial cell (HPMC) is another source of NGAL during peritonitis. NGAL was specifically induced in HPMC by IL-1beta. Incubation of HPMC with recombinant NGAL reversed the transforming growth factor-beta-induced up-regulation of Snail and vimentin but rescued the down-regulation of E-cadherin. Our data suggest that NGAL may exert a protective effect in modulating the epithelial-to-mesenchymal transition activated following peritonitis.

Topics: Acute-Phase Proteins; Adult; Ambulatory Care; Biomarkers; Cadherins; Cell Movement; Cells, Cultured; Early Diagnosis; Gene Expression Regulation; Humans; Interleukin-1beta; Kidney Failure, Chronic; Lipocalin-2; Lipocalins; Male; Middle Aged; Neutrophils; Peritoneal Dialysis; Peritonitis; Proto-Oncogene Proteins; Snail Family Transcription Factors; Streptococcal Infections; Streptococcus; Transcription Factors; Transforming Growth Factor beta; Vimentin

Peritoneal dialysis is an alternative treatment of patients with end-stage renal disease. Sclerosing encapsulating peritonitis is a life-threatening complication of continuous ambulatory peritoneal dialysis. The aim of the present study was to evaluate the effect of thalidomide, which is used for the treatment of various inflammatory and autoimmune diseases, on the development of sclerosing encapsulating peritonitis induced by chlorhexidine gluconate (CG). A peritoneal fibrosis model was established using rats treated intraperitoneally with injections of CG. Thalidomide was administered orally at a dose of 100 mg/kg three times per week. When compared with CG-treated rats, thalidomide (100 mg/kg orally)-treated mice subjected to CG-induced peritoneal fibrosis experienced a significantly lower rate in the extent and severity of histological signs of peritoneal injury. Thalidomide also caused a substantial reduction of 1) the rise in myeloperoxidase activity (mucosa); 2) the expression in the tissue of TNF-alpha, IL-1beta, transforming growth factor-beta, and vascular endothelial growth factor; 3) the increase in staining (immunohistochemistry) for nitrotyrosine and for poly(ADP ribose), as well as 4) the nuclear factor-kappaB activation caused by CG in the peritoneum. Thus, thalidomide treatment reduces the degree of peritoneal fibrosis caused by CG. We propose that this evidence may help clarify the potential therapeutic actions of thalidomide in patients with peritoneal fibrosis.

Topics: Animals; Anti-Infective Agents; Blotting, Western; Chlorhexidine; Disease Models, Animal; Enzyme Activation; Gene Expression Regulation; I-kappa B Proteins; Immunohistochemistry; Immunosuppressive Agents; NF-kappa B; Nitric Oxide Synthase Type II; Peritonitis; Peroxidase; Poly Adenosine Diphosphate Ribose; Rats; Thalidomide; Thiobarbituric Acid Reactive Substances; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Tyrosine; Vascular Endothelial Growth Factor A

It has been well demonstrated that TNF-alpha is integral to the pathogenesis of multiple organ dysfunction syndrome (MODS). In this study, we investigate the effects of etanercept (10 mg/kg, s.c.), a specific TNF-alpha-soluble inhibitor, on the acute phase and late mortality in a murine model of MODS of nonseptic origin induced by zymosan (500 mg/kg, suspended in saline solution, i.p.). Etanercept was administered 1 h after the injection of zymosan. Animals were killed after 18 h. In another set of experiments, mice were monitored for systemic toxicity, loss of body weight, and mortality for 12 days. Sham-treated and TNF receptor 1 (TNFR1)-deficient animals were used as control. Treatment of mice with Etanercept and TNFR1 gene deletion decreased the peritoneal exudation and the migration of neutrophils caused by zymosan. In addition, pharmacological and genetic neutralization of TNF-alpha attenuated pancreas and ileum injury (histology), the increase in myeloperoxidase activity in the ileum and in the lung, and the formation of TNF-alpha and IL-1beta. Immunohistochemical analysis for TNF-alpha, transforming growth factor beta, and vascular endothelial growth factor revealed a positive staining in pancreas and ileum sections. The degree of immunostaining was markedly reduced after etanercept treatment and in TNFR1 knockout mice. Furthermore, TNF-alpha neutralization decreased the potent apoptotic stimulus induced by zymosan. All of these findings ultimately led to an amelioration of organ functions at 18 h and to a better survival rate at 12 days. Therefore, we demonstrate that etanercept reduces acute tissue injury and mortality associated to MODS of nonseptic origin in mice.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; bcl-2-Associated X Protein; Etanercept; Fas Ligand Protein; Immunoglobulin G; Inflammation; Mice; Mice, Knockout; Multiple Organ Failure; Peritonitis; Proto-Oncogene Proteins c-bcl-2; Receptors, Tumor Necrosis Factor; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A; Zymosan

Lymphangiogenesis is induced by various growth factors, including VEGF-C. Although TGF-beta plays crucial roles in angiogenesis, the roles of TGF-beta signaling in lymphangiogenesis are unknown. We show here that TGF-beta transduced signals in human dermal lymphatic microvascular endothelial cells (HDLECs) and inhibited the proliferation, cord formation, and migration toward VEGF-C of HDLECs. Expression of lymphatic endothelial cell (LEC) markers, including LYVE-1 and Prox1 in HDLECs, as well as early lymph vessel development in mouse embryonic stem cells in the presence of VEGF-A and C, were repressed by TGF-beta but were induced by TGF-beta type I receptor (TbetaR-I) inhibitor. Moreover, inhibition of endogenous TGF-beta signaling by TbetaR-I inhibitor accelerated lymphangiogenesis in a mouse model of chronic peritonitis. Lymphangiogenesis was also induced by TbetaR-I inhibitor in the presence of VEGF-C in pancreatic adenocarcinoma xenograft models inoculated in nude mice. These findings suggest that TGF-beta transduces signals in LECs and plays an important role in the regulation of lymphangiogenesis in vivo.

Topics: Animals; Cell Line; Disease Models, Animal; Lymphangiogenesis; Mice; Mice, Nude; Pancreatic Neoplasms; Peritonitis; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptors, Transforming Growth Factor beta; Signal Transduction; Transforming Growth Factor beta; Vascular Endothelial Growth Factor C

Monosodium urate monohydrate (MSU) crystals have remarkable inflammatory potential. However, gouty inflammation is spontaneously self-limited, an occurrence recognized since antiquity. Gouty synovitis is driven and sustained by neutrophil influx. Importantly, macrophage phagocytosis of apoptotic (but not necrotic) neutrophils is antiinflammatory. Therefore, we tested the hypothesis that efficient clearance of apoptotic neutrophils by macrophages is one of the factors that restrains the progression of gouty inflammation. Macrophage expression of transglutaminase 2 (TG2), a multifunctional protein with reciprocally regulated transamidation and purine nucleotide-binding activities, promotes apoptotic leukocyte uptake. In this study, we tested the specific role of macrophage TG2 expression in MSU crystal-induced inflammation.. We studied MSU crystal-induced peritonitis in TG2-/- and congenic TG2+/+ mice. We also studied the effects of TG2 on apoptotic cell uptake by cultured macrophages.. TG2-/- mice demonstrated more progressive neutrophilic accumulation than did TG2+/+ mice, which was associated with delayed clearance of apoptotic neutrophils during MSU crystal-induced peritonitis. We observed defective phagocytosis of apoptotic leukocytes by TG2-/- peritoneal macrophages, which was corrected by soluble extracellular TG2. Transamidation catalytic activity of TG2 was not required to mediate macrophage uptake of apoptotic leukocytes. In contrast, the TG2 nucleotide binding site residue K173 was critical for this TG2 function. TG2 bound to GDP, ADP, or ATP (but not to GTP) rescued defective apoptotic leukocyte uptake by TG2-/- macrophages.. Enhancement of apoptotic neutrophil uptake by macrophage-derived TG2 restrains gout-like neutrophilic peritoneal inflammation. Differential binding of TG2 by purine nucleotides may contribute to clinical variability in the extent and duration of gouty inflammation.

Topics: Animals; Apoptosis; Cells, Cultured; Female; Gene Expression Regulation; Gout; GTP-Binding Proteins; Macrophages, Peritoneal; Male; Mice; Mice, Knockout; Neutrophils; Peritonitis; Phagocytosis; Protein Glutamine gamma Glutamyltransferase 2; Transforming Growth Factor beta; Transglutaminases; Uric Acid

Epithelial mesenchymal transition (EMT), a process involved in many growth and repair functions, has been identified in the peritoneal tissues of patients who undergo peritoneal dialysis. The sequence of changes in gene regulation and cellular events associated with EMT after TGF-beta1-induced peritoneal fibrosis is reported. Sprague-Dawley rats received an intraperitoneal injection of an adenovirus vector that transfers active TGF-beta1 (AdTGF-beta1) or control adenovirus, AdDL. Animals were killed 0 to 21 days after infection. Peritoneal effluent and tissue were analyzed for markers of EMT. In the animals that were treated with AdTGF-beta1, an increase in expression of genes associated with EMT and fibrosis, such as type I collagen A2, alpha-smooth muscle actin, and the zinc finger regulatory protein Snail, was identified. Transition of mesothelial cells 4 to 7 d after infection, with appearance of epithelial cells in the submesothelial zone 7 to 14 d after exposure to AdTGF-beta1, was demonstrated. This phase was associated with disruption of the basement membrane and increased expression of matrix metalloproteinase 2. By 14 to 21 d after infection, there was evidence of restoration of normal submesothelial architecture. These findings suggest that EMT occurs in vivo after TGF-beta1 overexpression in the peritoneum. Cellular changes and gene regulation associated with EMT are evident throughout the fibrogenic process and are not limited to early time points. This further supports the central role of TGF-beta1 in peritoneal fibrosis and provides an important model to study the sequence of events involved in TGF-beta1-induced EMT.

Topics: Animals; Biomarkers; Biopsy, Needle; Blotting, Western; Cells, Cultured; Disease Models, Animal; Epithelial Cells; Female; Immunohistochemistry; Injections, Intraperitoneal; Mesoderm; Peritoneum; Peritonitis; Polymerase Chain Reaction; Probability; Random Allocation; Rats; Rats, Sprague-Dawley; Reference Values; RNA, Messenger; Sensitivity and Specificity; Transforming Growth Factor beta; Transforming Growth Factor beta1

The peritoneal immune compartment is a microenvironment with a particular T-cell repertoire and susceptible to local inflammation. To clarify the role of T lymphocytes in peritoneal immunity, the changes in T-cell subpopulations in peritoneal dialysis effluents (PDEs), and their influence on the response to the treatment of peritonitis and on its prognosis were studied in patients undergoing long-term, continuous ambulatory peritoneal dialysis (CAPD).. A cohort of 36 patients treated with CAPD and who had histories of peritonitis were divided into a group with rapid and a group with delayed response to antibiotics, and were followed for 3 years. CD4/CD8 T-cell ratios, T-cell cytokine mRNA expression patterns and transforming growth factor-beta1 (TGF-beta1) concentrations were examined in PDE during bouts of peritonitis. The change in 4 h D/P creatinine during the peritoneal equilibration test (PET) between year 0 and year 3 was expressed as deltaD/P creatinine.. The serial changes in T-cell subsets in PDE during peritonitis showed two patterns: (i) pattern 1, manifest as a progressive increase in the CD4/CD8 ratio, and associated with a rapid response to treatment; and (ii) pattern 2, manifest as a progressive decrease in the CD4/CD8 ratio, and associated with a delayed response to treatment. The major T-cell phenotypes in PDE during peritonitis were Th1-CD4(+) and Tc2-CD8(+), determined by cloning techniques, RT-PCR and double immunofluorescence staining. TGF-beta1 in the effluent was undetectable in pattern 1 after 7-8 days, but remained detectable at 2 weeks in pattern 2. Pattern 2 patients had a significantly greater decrease (deltaD/P creatinine: -0.198+/-0.086) in solute transport than pattern 1 patients (deltaD/P creatinine: -0.036+/-0.077, P<0.05).. These results suggest that a progressive decrease of the CD4/CD8 ratio in PDE correlates with a persistent expression of TGF-beta1, and plays a pathogenetic role in the evolution of peritonitis, PET deterioration and peritoneal fibrosis. Therefore, patterns of CD4/CD8 T-cell ratio in PDE may predict clinical outcomes of peritonitis in CAPD patients.

Topics: Adult; Biomarkers; CD4 Antigens; CD4-CD8 Ratio; CD8 Antigens; Cytokines; Female; Fluorescent Antibody Technique; Humans; Kidney Failure, Chronic; Male; Microscopy, Fluorescence; Middle Aged; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; RNA, Messenger; T-Lymphocyte Subsets; Transforming Growth Factor beta

Connective tissue growth factor (CTGF) is a fibrogenic cytokine that is highly expressed in wound healing and fibrotic lesions. The role of transforming growth factor-beta (TGF-beta) in fibrosis is well documented, and the emerging understanding that its fibrogenic actions are mediated through CTGF has provided an attractive target molecule for the modulation of matrix overproduction in fibrotic disease. The involvement of CTGF in the pathogenesis of peritoneal membrane fibrosis in peritoneal dialysis (PD) patients has not been investigated, and so the aim of this study was to ascertain whether CTGF is produced in the peritoneal cavity of PD patients and to investigate its regulation by cytokines.. Reverse transcription-polymerase chain reaction (RT-PCR), Northern blotting, and Western blotting were used to study CTGF expression by cultured human peritoneal mesothelial cells (HPMC) from peritoneal dialysis patients. Western blotting was used to detect CTGF expression in spent peritoneal dialysate from patients with and without peritonitis.. RT-PCR analysis demonstrated the expression of CTGF mRNA in cultured primay HPMCs isolated from spent peritoneal effluent. The production of the major 36 to 38 kD CTGF protein doublet by HPMC in addition to a 23 to 25 kD proteolytically processed form was confirmed by Western blotting. Several molecular weight forms of CTGF (18 to 38 kD) were also detected by Western blotting in peritoneal dialysate, with levels markedly elevated during episodes of peritonitis. Northern and Western blot analysis revealed that CTGF mRNA and protein production by HPMC was up-regulated by TGF-beta, with mRNA levels significantly increasing above the control (P < 0.01). In contrast, platelet-derived growth factor (PDGF), epidermal growth factor (EGF), and tumor necrosis factor-alpha (TNF-alpha) had no measurable effects on CTGF mRNA expression.. These results are the first to demonstrate the production of CTGF by HPMC and its presence in the peritoneal cavity of PD patients. The marked increase in CTGF levels by factors implicated in the development of peritoneal membrane fibrosis suggests its involvement in the underlying pathophysiologic mechanism(s).

Topics: Base Sequence; Blotting, Northern; Blotting, Western; Case-Control Studies; Connective Tissue Growth Factor; Humans; Immediate-Early Proteins; Intercellular Signaling Peptides and Proteins; Molecular Sequence Data; Peritoneal Dialysis; Peritoneum; Peritonitis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transforming Growth Factor beta; Up-Regulation

Chronic peritoneal dialysis (CPD) is the most commonly used method of pediatric dialysis. The preservation of peritoneal membrane function is essential for successful peritoneal dialysis. Two main factors are responsible for long-term loss of membrane function. Peritonitis, the major complication of CAPD, can be life-threatening and lead to more rapid failure of the technique. The other factor is continuous exposure of peritoneal membrane to bioincompatible dialysis solutions. To investigate the role of repeated peritonitis on peritoneal membrane function, we performed a retrospective study to elucidate the association between peritonitis episodes and peritoneal membrane solute transport characteristics.. From 1996-2000, 32 pediatric peritoneal dialysis patients were included in this study. According to the peritonitis occurrence frequency, 8 patients were divided into HPO group (peritonitis occurrence rate > or = 5 times/year), and 24 patients were divided into LPO group (peritonitis occurrence rate < or = 1 time/year). The mean age of study patients was 13.95 +/- 5.27 years. The peritoneal equilibration test was performed to evaluate the peritoneal membrane dialyzing function.. The mean duration of peritoneal dialysis was 3.31 +/- 1.08 years. The change of peritoneal solute transport (AD/P), computed by subtraction of 4-hour D/P at baseline PET study from that at the last follow-up PET study, showed significant difference (p < 0.05) between HPO (-0.234 + 0.074) and LPO (-0.040 +/- 0.079) groups of children. There was also a significant correlation between repeated peritonitis occurrence and PET deterioration(p < 0.05). The relative risk was 2.63.. Children with frequent peritonitis occurrence have significant decreasing peritoneal solute transport and decreasing PET scaling in follow-up period.

Topics: Adolescent; Adult; Child; Child, Preschool; Female; Humans; Kidney Failure, Chronic; Male; Peritoneal Dialysis, Continuous Ambulatory; Peritoneum; Peritonitis; Transforming Growth Factor beta; Transforming Growth Factor beta1

The bioincompatibility of dialysis solutions and recurrent episodes of peritonitis may alter peritoneal function. Cytokines and growth factors may play a role in inflammatory and fibrotic processes. We therefore investigated whether there is a correlation between peritoneal function and excretion of cytokines and other soluble factors in dialysate.. A personal dialysis capacity test was performed in 40 stable peritoneal dialysis patients. Tumour necrosis factor-alpha, interleukin-6 (IL-6), hyaluronan, soluble intercellular cell adhesion molecule-1 and transforming growth factor-beta1 were analysed from overnight and 24-h dialysates during the test.. We found little evidence for a direct correlation between cytokines and other soluble factors in dialysate and dialysis adequacy. There was, however, a strong correlation between the measured soluble factors and characteristics of the peritoneal membrane. Furthermore, IL-6 correlated with the number of previous episodes of peritonitis.. Soluble factors in dialysate may indicate ongoing inflammatory processes in the peritoneal membrane, which may gradually lead to alterations in peritoneal function.

Topics: Adult; Aged; Cytokines; Dialysis Solutions; Female; Humans; Hyaluronic Acid; Intercellular Adhesion Molecule-1; Interleukin-6; Male; Middle Aged; Peritoneum; Peritonitis; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha

Infection with Trypanosoma cruzi causes a strong inflammatory reaction at the inoculation site and, later, in the myocardium. The present study investigates the role of cytokines as modulators of T. cruzi-induced chemokine expression in vivo and in vitro. In macrophage cultures, although the stimulation with interferon (IFN)-gamma increases the expression of IP-10, it blocks KC expression. Tumor necrosis factor (TNF)-alpha, on the other hand, potentiates KC, IP-10, macrophage inflammatory protein-1alpha, and JE/monocyte chemotatic protein-1 expression. Interleukin-10 and transforming growth factor-beta inhibited almost all chemokines tested. The role of IFN-gamma and TNF-alpha in chemokine modulation during infection was investigated in T. cruzi-infected IFN-gamma-deficient (GKO) or TNF-R1/p55-deficient (p55-/-) mice. The expression of chemokines detected in the inoculation site correlated with the infiltrating cell type observed. Although GKO mice had a delayed and intense neutrophilic infiltrate correlating with the expression of KC and macrophage inflammatory protein-2, none of the above was observed in p55-/- mice. The detection of infiltrating T cells, Mig, and IP-10 in the myocardium was observed in wild-type and p55-/-, but not in GKO mice. Together, these results suggest that the regulatory roles of IFN-gamma and TNF-alpha on chemokine expression may play a crucial role in the modulation of the inflammatory response during T. cruzi infection and mediate resistance to infection.

Topics: Animals; Antigens, CD; Cell Movement; Chagas Disease; Chemokine CXCL10; Chemokine CXCL9; Chemokines; Chemokines, CXC; Female; Immunophenotyping; Interferon-gamma; Interleukin-10; Lymphocytes; Macrophages; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Myocardium; Peritonitis; Receptors, Tumor Necrosis Factor; Receptors, Tumor Necrosis Factor, Type I; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Continuous ambulatory peritoneal dialysis (CAPD) has emerged as an important dialysis treatment modality worldwide. One of the major complications is bacterial peritonitis, which may result in subsequent technique failure because of loss of peritoneal clearance or peritoneal fibrosis. Bacterial peritonitis leads to the release of proinflammatory cytokines from resident and infiltrating cells in the peritoneal cavity. We studied 35 patients undergoing CAPD with acute bacterial peritonitis. All patients treated with antibiotics for 2 weeks after the clinical diagnosis of peritonitis had a good recovery. Peritoneal dialysate effluent (PDE) was collected on days 1, 3, 5, 10, 21, and 42 after the start of treatment. Cell populations were monitored by flow cytometry. PDE levels of interleukin-1beta (IL-1), IL-6, transforming growth factor-beta (TGF-beta), and basic fibroblast growth factor (FGF) were measured by enzyme-linked immunosorbent assay. Gene transcription of TGF-beta in macrophages from PDE was measured by quantitative polymerase chain reaction. Bacterial peritonitis was associated with a sharp increase in total cell and neutrophil counts (400-fold) in PDE up to 3 weeks after peritonitis despite clinical remission (P < 0.0001). There was an increased absolute number of macrophages during the first 3 weeks despite the reduced percentage of macrophages among total cells in PDE compared with noninfective PDE. There was a progressive increase in the percentage of mesothelial cells or dead cells in the total cell population in PDE over the entire 6-week period. PDE levels of IL-1, IL-6, TGF-beta, and FGF increased markedly on day 1 before their levels decreased gradually. PDE levels of these cytokines or growth factors were significantly greater than those in noninfective PDE (n = 76) throughout the study period (P < 0.01). Similarly, TGF-beta complementary DNA (cDNA) molecules per macrophage were significantly greater than those of macrophages in noninfective PDE throughout this period (P < 0.01). There was no significant correlation between PDE levels of TGF-beta and TGF-beta cDNA molecules per macrophage, suggesting that peritoneal macrophages are not the only source of TGF-beta in PDE. We conclude there is an active release of proinflammatory cytokines and sclerogenic growth factors through at least 6 weeks despite apparent clinical remission of peritonitis. The peritoneal cytokine networks after peritonitis may potentially affect the physiological pro

Topics: Bacterial Infections; Cytokines; DNA, Complementary; Female; Fibroblast Growth Factor 2; Fibrosis; Flow Cytometry; Humans; Interleukin-1; Interleukin-6; Macrophages, Peritoneal; Male; Middle Aged; Peritoneal Dialysis, Continuous Ambulatory; Peritoneal Diseases; Peritonitis; Polymerase Chain Reaction; Transforming Growth Factor beta

Long-term influence of continuous ambulatory peritoneal dialysis (CAPD) on concentrations of transforming growth factor beta1 (TGF-beta1) and basic fibroblast growth factor (bFGF) in the peritoneal effluent, and the effect of peritonitis on these cytokines were investigated. TGF-beta1 and bFGF were assayed in effluent samples from dialysate bags collected during the initial week of treatment with CAPD and at 5 months. To determine the effect of peritonitis, dialysate bags were collected on admission to the hospital and on days 3 and 10 and also during non-infected steady state. Serum was drawn prior to infection and on days 1 and 10. TGF-beta1 increased more than threefold during the longitudinal follow-up period, median concentrations of 35 pg/ml to 106 pg/ml (P<0.05). No change in bFGF was seen during this initial 5 months. TGF-beta1 was increased on the first day of peritonitis (median concentration 169 pg/ml) and reached its maximum on day 3 of infection, (median concentration 216 pg/ml) (P<0.05 vs non-infected state, median concentration 39 pg/ml). Basic FGF reached a maximum on day three of infection (median concentration 7.7 pg/ml; P=0.01 vs non-infected state) and then slowly declined. In conclusion, TGF-beta1 is influenced by CAPD treatment per se, and together with bFGF is increased during peritonitis, indicating its importance in the peritoneum and its potential involvement in the development of tissue fibrosis and eventually ultrafiltration failure.

Topics: Adult; Aged; Corynebacterium; Dialysis Solutions; Escherichia coli; Female; Fibroblast Growth Factor 2; Fibrosis; Humans; Interleukin-13; Interleukin-2; Longitudinal Studies; Male; Middle Aged; Peritoneal Dialysis, Continuous Ambulatory; Peritoneal Diseases; Peritonitis; Staphylococcus aureus; Staphylococcus epidermidis; Streptococcus sanguis; Transforming Growth Factor beta; Uremia

One of main challenges of peritoneal dialysis (PD) is the functional and vital long-term stability of the peritoneal membrane. Few longitudinal and controlled studies on peritoneal function have been published, and the results are somewhat contradictory. We have performed a longitudinal study with 90 patients. The overall analysis has shown that creatinine mass transfer coefficient (MTC) significantly increases and ultrafiltration (UF) capacity decreases over time. Nevertheless, urea MTC remained unaltered and MTC ratios significantly decreased after the third year. Subsequently, we examined the clinical outcomes and identified 19 patients who required peritoneal resting periods for Type I UF failure and 71 patients who did not require such a procedure. The latter patients did not show any significant functional change over time, whereas the former 19 patients showed an increase of peritoneal creatinine transport and a loss of UF capacity. These data corroborate changes in long-term peritoneal function in approximately 20% of PD patients. These changes consist of an increase in effective exchange area, peritoneal permeability, or both, accompanied by signs suggestive of mesothelial regenerative capacity loss. Infectious peritoneal injuries, especially appearing during late PD periods, are deleterious to the peritoneum. The remainder of the functional-structural changes are related to the effects of currently used dialysate. Early diagnosis, preemptive, and therapeutic measures should permit better management of long-term PD patients. The particular response to these injuries has individual characteristics that when addressed permit PD to be used long-term.

Topics: Analysis of Variance; Creatinine; Dialysis Solutions; Glycosylation; Humans; Longitudinal Studies; Peritoneal Dialysis; Peritoneum; Peritonitis; Prognosis; Sclerosis; Transforming Growth Factor beta; Ultrafiltration

The efficiency of continuous ambulatory peritoneal dialysis depends on the permeability of the peritoneal membrane. Peritoneal fibrosis (PF) causes loss of the dialytic function. Several studies have indicated that PF is closely related to the proliferation of peritoneal fibroblasts and the deposition of extracellular matrix. Transforming growth factor-beta 1 (TGF-beta1) plays a major role in stimulating extracellular matrix deposition. Frequent peritonitis occurrence may cause persistent TGF-beta1 mRNA expression. In an attempt to search for a factor related to PF, we designed a longitudinal study to measure TGF-beta1 levels in dialysate and TGF-beta1 mRNA expression in peritoneal mononuclear cells from peritoneal dialysate before onset, once a week during peritonitis, and after peritonitis in high and low peritonitis occurrence (HPO and LPO) patients. Fifteen patients with a LPO rate and 5 patients with a HPO rate were followed up longitudinally. Meanwhile, TGF-beta1 levels and TGF-beta1 mRNA expression were augmented in peritoneal dialysate effluents before, during, and after the episodes of peritonitis. The peritoneal permeability was evaluated by the peritoneal equilibration test. The results revealed that in the LPO group, TGF-beta1 and TGF-beta1 mRNA were detectable at early stages of peritonitis, but the levels decreased rapidly and were undetectable 2 weeks after peritonitis. On the other hand, in the HPO group, TGF-beta1 and TGF-beta1 mRNA persisted for a long time. We could detect TGF-beta1 and TGF-beta1 mRNA in dialysate effluents and peritoneal mononuclear cells even 2, 3, and 4 weeks after episodes of peritonitis. When compared with that of first or second episode of peritonitis, the peritoneal function evaluated with the peritoneal equilibration test was found to obviously deteriorate during the third episode of peritonitis. These findings were confirmed by an in situ hybridization technique to evaluate the relationship between TGF-beta1 mRNA expression and PF from biopsied peritoneal specimens. These findings suggest that the high TGF-beta1 levels in the dialysate are related to an increased expression of TGF-beta1 in the peritoneum. Thus, the persistent TGF-beta1 expression in the peritoneum may serve as a useful parameter in predicting PF in continuous ambulatory peritoneal dialysis patients with frequent peritonitis occurrence.

Topics: Adolescent; Adult; Blotting, Northern; Child; Creatinine; Dialysis Solutions; Fibrosis; Glucose; Humans; In Situ Hybridization; Middle Aged; Peritoneal Dialysis, Continuous Ambulatory; Peritoneum; Peritonitis; RNA, Messenger; Transforming Growth Factor beta

The efficiency of continuous ambulatory peritoneal dialysis (CAPD) depends on the permeability of the peritoneal membrane. Peritoneal fibrosis (PF) causes the loss of dialytic function. Several studies have indicated that PF is closely related to the proliferation of peritoneal fibroblasts and the deposition of extracellular matrix (ECM). Transforming growth factor beta 1 (TGF beta 1) plays a major role in stimulating ECM deposition. Frequent peritonitis occurrence may cause persistent TGF beta 1 mRNA expression. In an attempt to search for a factor related to PF, we designed a longitudinal study to measure TGF beta 1 levels in dialysate and TGF beta 1 mRNA expression in peritoneal mononuclear cells (PMNCs) from peritoneal dialysate before, at the onset of and once a week during peritonitis and after peritonitis in patients with high peritonitis occurrence (HPO) and patients with low peritonitis occurrence (LPO). Fifteen patients with a LPO rate and 5 patients with a HPO rate were followed up longitudinally. Meanwhile, TGF beta 1 levels and TGF beta 1 mRNA expression were augmented in peritoneal dialytic fluid before, during, and after the episodes of peritonitis. Peritoneal permeability was evaluated by the peritoneal equilibration test (PET). The results revealed that in the LPO group, TGF beta 1 and TGF beta 1 mRNA were detectable at early stages of peritonitis, but the levels decreased rapidly and were undetectable 2 weeks after peritonitis. On the other hand, in the HPO group, TGF beta 1 and TGF beta 1 mRNA persisted for a long time. We could detect TGF beta 1 and TGF beta 1 mRNA in dialytic fluid and PMNCs even 2, 3, and 4 weeks after episodes of peritonitis. When compared with that of the first or second episode of peritonitis, peritoneal function evaluated with the PET was found to obviously deteriorate at the third episode of peritonitis. These findings were confirmed by an in situ hybridization technique to evaluate the relationship between TGF beta 1 mRNA expression and PF from biopsied peritoneal specimens. These findings suggest that the high TGF beta 1 levels in the dialysate are related to an increased expression of TGF beta 1 in the peritoneum. Persistent TGF beta 1 expression in the peritoneum may serve as a useful parameter in predicting PF in CAPD patients with frequent peritonitis occurrence.

Topics: Adolescent; Adult; Blotting, Northern; Child; Creatinine; Dialysis Solutions; Fibrosis; Glucose; Humans; In Situ Hybridization; Leukocytes, Mononuclear; Longitudinal Studies; Middle Aged; Peritoneal Dialysis, Continuous Ambulatory; Peritoneum; Peritonitis; Permeability; Recurrence; RNA, Messenger; Transforming Growth Factor beta

We have investigated the role of human mesothelium in an in vitro model of peritonitis on the monocyte adherence to and migration across monolayers of peritoneal mesothelial cells. Monocytes adhere avidly to non-activated mesothelial cell monolayers; however, migration in this situation was minimal. Prestimulation of the monolayers with II-1 beta did not alter these results. Anti-CD18 and anti-VLA-4 mAbs used in combination had an additive inhibitory effect on monocytes adherence to resting or IL-1 beta-pretreated mesothelial cells. MCP-1 and TGF-beta are secreted by mesothelial cells. Both have a modest role in mesothelium-induced monocyte chemotaxis: mAbs against these cytokines had an additive inhibitory effect on the chemotaxis induced by supernatant from 24-h prestimulated mesothelial cells. Our results indicate that the mesothelium itself has a limited role in the influx of monocytes into the peritoneal cavity during the onset of peritonitis.

Topics: Antibodies, Monoclonal; CD18 Antigens; Cell Adhesion; Cells, Cultured; Chemokine CCL2; Chemotaxis, Leukocyte; Culture Media, Conditioned; E-Selectin; Epithelium; Humans; Integrin alpha4beta1; Integrins; Intercellular Adhesion Molecule-1; Interleukin-1; Monocytes; Peritoneal Cavity; Peritonitis; Receptors, Lymphocyte Homing; Transforming Growth Factor beta

Elicited guinea pig macrophages collected from inflammatory peritoneal exudate release soluble 14 kDa phospholipase A2 (PLA2) and prostaglandin E2 (PGE2) into surrounding media during culture (Marshall et al. (1994) J. Lipid Med. 10, 295-313). The effect of transformation growth factor beta 1 (TGF beta), an immunoregulatory growth factor, was examined in this system. Exposure of cultured macrophages to TGF beta reduced both the activity and protein levels of 14 kDa PLA2 measured in conditioned media. This inhibition occurred within the first 6-8 h, was prevalent through 72 h of exposure and was dependent on TGF beta concentration. The reduction, however, never reached more than 40-60%. Evaluation of the cellular PLA2 activity confirmed the existence of an immunologically-related 14 kDa PLA2 (ELISA) in the particulate fraction and an 85 kDa PLA2 (Western analysis) in the cytosol. Exposure to TGF beta halved the particulate activity and protein levels of 14 kDa PLA2 which was consistent with the reduction in the secreted form. Alternatively, TGF beta induced an increase in cytosolic 85 kDa PLA2 (activity and protein) which was not apparent until 12 h and significant at 20-24 h of exposure. This demonstrates that TGF beta differentially regulates the production of these two enzymes. Despite this, neither PGE2 synthesis nor the up-regulated cyclooxygenase -II were altered by TGF beta treatment suggesting that maximal prostanoid synthesis had been reached.

Topics: Animals; Dinoprostone; Guinea Pigs; Isoenzymes; Macrophage Activation; Macrophages, Peritoneal; Male; Peritonitis; Phospholipases A; Phospholipases A2; Prostaglandin-Endoperoxide Synthases; Transforming Growth Factor beta