Inhibition of cytokine synthesis by peritoneal dialysate persists throughout the CAPD cycle.

The current study focused on the effect of continuous ambulatory peritoneal dialysis (CAPD) dialysate obtained following different intraperitoneal dwell periods on the release of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF alpha) from mononuclear leukocytes (PBMC). Aliquots of 5 x 10(6)/ml healthy peripheral PBMC were exposed to fresh or spent CAPD dialysate (10-240 min of intra-peritoneal dwell) and stimulated with Escherichia coli endotoxin (10 micrograms/ml, 2h). IL-6 and TNF alpha in cell supernatants were determined by specific enzyme immunoassays. Control PBMC in physiological buffer released 361 +/- 70 pg/ml IL-6 and 717 +/- 147 pg/ml TNF alpha (mean +/- SEM, n = 8), whereas exposure to fresh dialysis fluids severely suppressed cytokine release from PBMC (less than 30 pg/ml IL-6 and less than 15 pg/ml TNF alpha). A significant inhibition of IL-6 and TNF alpha release was also observed in PBMC exposed to spent dialysate. The inhibitory capacity of the spent fluids was pronounced with increasing intra-peritoneal dwell time (10 min: 183 +/- 45 pg/ml IL-6 and 538 +/- 109 pg/ml TNF alpha; 240 min: 26 +/- 5 pg/ml IL-6 and 105 +/- 30 pg/ml TNF alpha; mean +/- SEM, n = 16). These data indicate that the impairment of cell responsiveness following exposure of PBMC to peritoneal dialysate is not restricted to the unused fluids, but is also observed following intra-peritoneal equilibration. Moreover, our findings suggest the presence of cytokine inhibitory factors in the peritoneal dialysate of CAPD patients which appear to accumulate in the peritoneal effluent during the CAPD cycle.     

Patterns of CD4/CD8 T-cell ratio in dialysis effluents predict the long-term outcome of peritonitis in patients undergoing peritoneal dialysis.

BACKGROUND: 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). METHODS: 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. RESULTS: 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). CONCLUSIONS: 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.     

Plasma interleukin-6 levels in continuous ambulatory peritoneal dialysis and hemodialysis patients.

Plasma levels of interleukin-6 (IL-6), a cytokine known to be involved in lymphocyte activation and in inflammation, were studied in 10 normal volunteers, 21 continuous ambulatory peritoneal dialysis (CAPD) patients and 41 hemodialysis patients. Plasma IL-6 levels in hemodialysis patients were significantly higher than those in normal volunteers and CAPD patients (p less than 0.05). The means of plasma IL-6 concentrations before and after hemodialysis did not change significantly. While IL-6 in peritoneal dialysate was detectable in only 3 of the 21 CAPD patients without peritonitis, it was extremely high in 2 patients with bacterial peritonitis. IL-6 levels decreased as peritonitis subsided.     

Plasma and dialysate IL-6 and VEGF concentrations are associated with high peritoneal solute transport rate.

BACKGROUND: It has been speculated that increased levels of circulating or intraperitoneal pro-inflammatory cytokines such as interleukin 6, and pro-angiogenic vascular endothelial growth factor (VEGF) may contribute to high peritoneal small-solute transport rate (PSTR) in continuous ambulatory peritoneal dialysis (CAPD) patients. In this study we evaluated possible relationships between plasma and dialysate IL-6 and VEGF levels and PSTR. METHODS: Forty CAPD patients (mean age+/-SD of 58+/-14 years) with no apparent inflammation process or disease, who had been on CAPD for 19+/-15 months (range 3-56 months) were included in the study. Peritoneal equilibration test (PET) was used to evaluate PSTR. Patients were divided into two groups: high-average and high transporters (H/A; D/P(creat)>/=0.65) and low-average and low transporters (L/A; D/P(creat)<0.64). Albumin and IgG clearances were used in the evaluation of permeability to larger solutes. Plasma and overnight dialysate levels of IL-6 and VEGF were measured. RESULTS: Plasma IL-6 (7.6 vs 4.3 pg/ml) and VEGF (342 vs 163 pg/ml) as well as dialysate IL-6 (174 vs 80 pg/ml) and VEGF (96 vs 69 pg/ml) levels were significantly higher in the H/A than in the L/A group. The dialysate appearance of IL-6 and VEGF correlated with D/P(creat), as well as with albumin and IgG clearances. Moreover, significant correlations were noted between dialysate IL-6 and dialysate VEGF levels. CONCLUSIONS: The findings of (i) increased plasma and dialysate levels of IL-6 and VEGF in the H/A group compared to the L/A group, (ii) an association between PSTR and both plasma and dialysate IL-6 and VEGF levels, and (iii) a significant correlation between dialysate IL-6 and VEGF concentrations suggest that inflammation, angiogenesis, and peritoneal transport may be interrelated and involved in the pathophysiology of high PSTR in CAPD patients. However, due to the cross-sectional design of this study, the cause and effect relationships between plasma and dialysate IL-6 and VEGF concentrations and high PSRT remain unclear.     

Subclinical hypothyroidism is linked to micro-inflammation and predicts death in continuous ambulatory peritoneal dialysis.

BACKGROUND: Low T3 is a frequent alteration in patients with ESRD. This derangement has been recently linked to inflammation in haemodialysis patients. Whether this association holds true in peritoneal dialysis patients has not been studied. METHODS: We investigated the relationship between low-grade inflammation [IL-6, C-reactive protein (CRP) and serum albumin levels] and free tri-iodothyronine (fT3) in a cohort of 41 CAPD patients (mean age, 66 years; M, 26; F, 15) without heart failure and inter-current illnesses. RESULTS: CAPD patients had lower fT3 levels (2.7 +/- 0.8 pg/ml) than healthy subjects (3.7 +/- 1.0 pg/ml, P < 0.001) of similar age. Free T3 levels were directly related to those of serum albumin (r = 0.52, P = 0.001) and inversely to IL-6 (r = -0.30, P = 0.05) and CRP (r = -0.54, P < 0.001). Age (r = -0.61, P < 0.001), haemoglobin levels (r = 0.32, P = 0.05) and diastolic blood pressure (r = 0.50, P = 0.001) were also related to fT3. In multiple regression models adjusting for all variables related to fT3, CRP and albumin were retained as independent correlates of fT3. During the follow-up (2.8 +/- 1.7 years) 27 patients died. Plasma fT3 levels were lower in patients who died (2.5 +/- 0.8 pg/ml) compared with survivors (3.3 +/- 0.5 pg/ml P = 0.001). In Cox analyses, fT3 was a significant predictor of mortality independent of the main traditional as well as non-traditional risk factors. CONCLUSIONS: The relationship between fT3, CRP and serum albumin suggests that inflammation-malnutrition might be involved in the low T3 syndrome in CAPD patients. Thyroid dysfunction might be implicated in the pathogenic pathway which links micro-inflammation to survival in PD patients.     

In vitro production of interleukin-1 receptor antagonist in chronic renal failure, CAPD and HD.

Dialysis-related symptoms are believed to be mediated, at least in part, by monocyte/macrophage-derived pro-inflammatory cytokines including interleukin-1 (IL-1) and tumor necrosis factor (TNF). Measuring the production of interleukin-1 receptor antagonist (IL-Ra), a naturally occurring inhibitor of IL-1, opens avenues to study the balance between these two cytokines in patients. We studied the cell content and production of IL-1 beta and IL-Ra by unstimulated and endotoxin- or IgG-stimulated peripheral blood mononuclear cells (PBMC) in undialyzed patients with chronic renal failure (CRF), patients on continuous ambulatory peritoneal dialysis (CAPD) and patients on chronic hemodialysis with reuse cuprophan membranes (HD), and compared them to healthy controls. IL-1 beta and IL-Ra were measured by specific radioimmunoassay. IL-1 beta was undetectable in freshly harvested PBMC from healthy controls, CRF, CAPD or HD. In contrast, the content of IL-Ra in HD patients (2828 +/- 466 pg/ml) was significantly higher than that in healthy controls (643 +/- 53 pg/ml, P < 0.01), CRF (1097 +/- 320 pg/ml, P < 0.01) or CAPD (1398 +/- 390 pg/ml, P < 0.05). In endotoxin-stimulated PBMC, IL-1 beta production by HD patients (9375 +/- 1687 pg/ml) was not significantly different from healthy controls (8429 +/- 1621 pg/ml). However, endotoxin-stimulated IL-Ra production by HD patients (32,350 +/- 8276 pg/ml) was greater than that from healthy controls (11,284 +/- 1250 pg/ml, P < 0.001), CRF (12,263 +/- 2680 pg/ml, P < 0.01) or CAPD patients (11,822 +/- 1797 pg/ml, P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)     

Serum and peritoneal fluid amylase levels in CAPD. Normal values and clinical usefulness

The mean serum amylase of 42 asymptomatic CAPD patients was elevated but was not significantly different from that of a group of chronic hemodialysis patients. Serum amylase levels in CAPD patients with peritonitis were not elevated with respect to asymptomatic patients. Amylase activity was not detectable in the peritoneal fluid of 38/42 asymptomatic patients and 6/13 peritonitis patients and was present at low levels in the other 11 patients. Patients with other abdominal conditions (pancreatitis, cholecystitis and small bowel perforation) had very marked elevations of serum and/or peritoneal fluid amylase which differentiated them from the asymptomatic and peritonitis patients. Although hyperamylasemia is common in asymptomatic CAPD patients and in those with peritonitis, measurement of serum and peritoneal fluid amylase levels is useful in the evaluation of CAPD patients presenting with abdominal symptoms.     

Plasma levels of human atrial natriuretic factor in patients treated by hemodialysis and continuous ambulatory peritoneal dialysis

We measured plasma levels of immunoreactive human atrial natriuretic factor (ANF) in chronic renal failure patients treated by hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD). Predialysis plasma ANF was significantly higher in HD patients (271.8 +/- 173.4 pg/ml) as compared to CAPD patients (81.8 +/- 80.5 pg/ml) and healthy subjects (31.5 +/- 19.8 pg/ml). Plasma volume was higher in HD patients than in CAPD patients. Plasma ANF and plasma volume showed a significant positive correlation. In HD patients, high plasma ANF value decreased significantly to a value comparable with that of CAPD patients after each dialysis. The removal rates of ANF by HD and CAPD were comparable. Ultrafiltration corresponding to 2% of body weight without dialysis also reduced plasma ANF. Thus, the difference in plasma ANF values between HD and CAPD patients seems to be mostly due to the difference in plasma volume, indicating that plasma ANF is sensitive to volume status even in chronic dialysis patients.     

Impact of dialysate interleukin-6 level on the alteration of peritoneal solute transport rate in peritoneal dialysis patients

Objective To evaluate the effect of dialysate interleukin-6 (IL-6), a marker of ongoing peritoneal inflammation, on the alteration of peritoneal solute transport rates (PSTRs) in stable continuous ambulatory peritoneal dialysis (CAPD) patients. Methods Atotal of 128 case of stable CAPD patients were enrolled in present study.IL-6 levels in the overnight effluent were determined by ELISA and IL-6 appearance rates (AR) were calculated. Mass transfer area coefficients of creatinine (MTACcr) were prospectively followed up. Logistic regression was used to examine the association between IL-6 AR and increased PSTRs. Results The MTACcr was significantly increased after 12 months follow-up[M(1/4，3/4), 6.40(4.70, 8.75) ml/min vs 7.14(5.59, 8.73) ml/min, P＜0.05]. Compared to the patients with stable PSTRs, the dialysate IL-6 AR in patients with increased PSTRs showed significantly higher [277.08(247.45, 349.53) pg/min vs 263.18 (69.94, 286.72) pg/min, P＜0.05]. Patients with increased PSTRs also had lower residual renal function [0.79(0, 2.12) ml/min vs 1.70 (0.39, 3.38) ml/min, P＜0.05], less urine volume [225(0, 600) ml/24 h vs 500(125, 900) ml/24 h, P＜0.05] and lower baseline MTACcr [5.48 (4.17, 7.42) ml/min vs 7.00(5.46, 9.76) ml/min, P＜0.05] when compared to their counterparts with stable PSTRs. Logistic analysis showed that high dialysate IL-6 AR and low baseline MTACcr were independent risk factors for increasing peritoneal solute transport rate (P＜0.05). Conclusion Intra-peritoneal inflammation significantly affects the alteration of PSTRs, and the dialysate IL-6 may be a predictor for increased PSTRs in PD patients.     

Peritonitis increases MMP-9 activity in peritoneal effluent from CAPD patients

Patients undergoing long-term continuous ambulatory peritoneal dialysis (CAPD) sometimes experience ultrafiltration failure. Mesothelial basement membrane thickening and the accumulation of submesothelial fibrotic tissue are common features of the diseased peritoneum. Peritonitis can lead to ultrafiltration failure, but the precise mechanism is not clear. The key enzymes in extracellular matrix (ECM) remodeling, namely matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs), are produced by human peritoneal mesothelial cells. Using peritoneal effluent from 13 CAPD patients with peritonitis and 7 noninfected CAPD control individuals, we examined MMP and TIMP activities by gelatin and reverse zymography. Latent and activated types of MMP-2 and -9, and TIMP-1 and -2 were identified in peritoneal effluent (from all CAPD patients). Levels of latent and activated type MMP-9, as well as of TIMP-1 activities were higher at the onset of peritonitis than either during the recovery phase of peritonitis and/ or in control individuals. Activated MMP-9 activity positively correlated with leukocyte numbers and IL-6 levels in peritoneal effluent. Activities of MMP-2 and TIMP-2 in peritoneal effluent did not change between the onset of peritonitis and recovery. We concluded that increased MMP-9 and TIMP-1 levels might be associated with peritoneal ECM remodeling during peritonitis.     

Proteomic analysis of peritoneal dialysate fluid in patients with dialysis-related peritonitis

The prognosis of uremia patients on continuous ambulatory peritoneal dialysis (CAPD) is related to frequent peritonitis rate. Frequent peritonitis will lead to peritoneum failure, making CAPD unfeasible. We have performed proteomic profiling of peritoneal dialysis effluent samples from a cross-section of CAPD patients with and without peritonitis in order to identify biomarkers of peritonitis. We performed 2D gel electrophoresis and surface-enhanced laser esorption/ionization time of flight mass spectrometry (SELDI-TOF MS) on peritoneal dialysis effluent from 16 subjects with peritonitis. A genetic algorithm search of principal component space revealed a group of a peak distinguishing peritonitis-positive subjects, with mass/charge (m/z) values of 11,117.4. Our analyses identified the peak at m/z 11,117.4 with an accuracy of 95% for classifying peritonitis. Mass spectrometric analysis of peritonitis PDE samples identified the 11,117.4 protein as beta2-microglobulin (B2M). Using an unbiased protein profiling approach, we have validated previously reported findings of B2M as a biomarker associated with CAPD peritonitis. Prospective studies are warranted to establish additional biomarkers that would be predictive of peritoneal dialysis peritonitis. Besides, extending the study to a larger number of patients with subgroup analyses may yield additional information of the peritoneal dialysate proteins in association with dialysis adequacy, residual renal function, nutritional status, and risk of peritoneal infection.     

Proteomic Analysis of Peritoneal Dialysate Fluid in Patients with Dialysis-Related Peritonitis

The prognosis of uremia patients on continuous ambulatory peritoneal dialysis (CAPD) is related to frequent peritonitis rate. Frequent peritonitis will lead to peritoneum failure, making CAPD unfeasible. We have performed proteomic profiling of peritoneal dialysis effluent samples from a cross-section of CAPD patients with and without peritonitis in order to identify biomarkers of peritonitis. We performed 2D gel electrophoresis and surface-enhanced laser esorption/ionization time of flight mass spectrometry (SELDI-TOF MS) on peritoneal dialysis effluent from 16 subjects with peritonitis. A genetic algorithm search of principal component space revealed a group of a peak distinguishing peritonitis-positive subjects, with mass/charge (m/z) values of 11,117.4. Our analyses identified the peak at m/z 11,117.4 with an accuracy of 95% for classifying peritonitis. Mass spectrometric analysis of peritonitis PDE samples identified the 11,117.4 protein as β2-microglobulin (B2M). Using an unbiased protein profiling approach, we have validated previously reported findings of B2M as a biomarker associated with CAPD peritonitis. Prospective studies are warranted to establish additional biomarkers that would be predictive of peritoneal dialysis peritonitis. Besides, extending the study to a larger number of patients with subgroup analyses may yield additional information of the peritoneal dialysate proteins in association with dialysis adequacy, residual renal function, nutritional status, and risk of peritoneal infection.     

Relationship between serum 25-hydroxycholecalciferol deficiency and the risk of peritoneal dialysis associated peritonitis

Objective To investigate the relationship between serum 25-hydroxycholecalciferol[25(OH)D3] deficiency and the risk of peritoneal dialysis associated peritonitis. Methods Baseline clinical data (before the peritoneal dialysis catheter insertion) of peritoneal dialysis patients treated with CAPD in the First Affiliated Hospital of Guangxi Medical University from May 1, 2013 to February 1, 2016 were retrospective analyzed. All the patients were followed-up until July 31, 2016. According to the baseline serum 25(OH)D3 levels, patients were divided into deficiency group (25(OH)D3＜15 ng/ml) and non deficiency group (25(OH)D3 ≥15 ng/ml), the baseline clinical data of the two groups were also analyzed. Kaplan-Meier method was used to compare the time-to-peritonitis of two groups. Cox proportional hazard model was used to analyze the relationship between the 25(OH)D3 deficiency and the risk of peritonitis. ROC curve was used to analyze the predictive value of the baseline serum 25(OH)D3 for the risk of PDAP in peritoneal dialysis patients. Results Compared with the 25(OH)D3 non deficiency group, 25(OH)D3 deficiency group had a significant increase incidence of peritonitis, high diastolic blood pressure and mean arterial pressure, but serum albumin, total serum protein decreased significantly (P＜0.05). Kaplan-Meier survival analysis showed that, compared with 25(OH)D3 non deficiency group, the time-to-peritonitis episode of patients with 25(OH)D3 deficiency were shorter (P＜0.05). Cox proportional hazard model showed that after adjusting for age, sex, hemoglobin, serum albumin, C-reactive protein, total Kt/V, eGFR, diabetes or not, 25(OH)D3 deficiency is the independent risk factor of peritoneal dialysis associated peritonitis (HR 5.247, 95%CI 1.180-23.340, P＜0.05). ROC curve showed the area under the curve that baseline serum 25(OH)D3 deficiency predict the occurrence of PDAP was 0.714, and the best cut-off point of baseline serum 25(OH)D3 was 11.35 ng/ml (sensitivity 75%, specificity 63%). Conclusions Peritoneal dialysis associated peritonitis occurred earlier in peritoneal dialysis patients whose baseline serum 25(OH)D3 deficiency. Baseline serum 25(OH)D3 deficiency is the independent risk factor of peritoneal dialysis associated peritonitis, which may predict the incidence of peritoneal dialysis associated peritonitis.     

Losses of 1,25- and 24,25-dihydroxycholecalciferol in the peritoneal fluid of patients treated with continuous ambulatory peritoneal dialysis

We measured peritoneal losses of the active vitamin D metabolites 1,25(OH)2D3 and 24,25(OH)2D3 in patients receiving continuous ambulatory peritoneal dialysis (CAPD). The serum concentration of 24,25(OH)2D3 was considerably lower than in hemodialysis patients. The serum concentration of 1,25(OH)2D3 was undetectable and rose to levels similar to those in hemodialysis patients only after loading with much higher oral doses of 1-alpha-vitamin D3 than those received by hemodialysis patients. Losses of both metabolites in peritoneal fluid were considerable, averaging approximately 6-8% of the plasma pool per day. These losses lead to low serum levels of these active vitamin D metabolites in CAPD patients, which may be an important factor in exacerbating renal osteodystrophy. Our results indicate the need for increased replacement doses of vitamin D metabolites in CAPD patients.     

Recurrent peritonitis: evidence for possible viral etiology.

A 45-year-old woman who was treated with continuous ambulatory peritoneal dialysis (CAPD) developed recurrent peritonitis characterized by cloudy effluents, elevated white blood cell (WBC) counts (predominantly lymphocytes), and negative culture results. This case report suggests that she may have had viral peritonitis as indicated by a positive viral culture, the presence of viral antibodies in serum and peritoneal dialysis effluent (PDE), hematological findings, and cell surface receptor studies. The possibility of a viral cause should be considered in patients with culture-negative peritonitis, especially if they do not respond to antibiotics.     

Interleukin-8 in chronic renal failure and dialysis patients

A total of 105 patients participated in this study, including10 with chronic glomerulonephritis with normal renal function(CGN patients), 36 uraemic patients (CRF patients), 19 continuousambulatory peritoneal dialysis patients (CAPD) without peritonitis,three CAPD patients with peritonitis, 37 patients undergoingchronic haemodialysis (HD) divided into short-term HD, 15 patients;medium-term HD, 12 patients; and long-term HD, 10 patients.IL-8 and two other proinflammatory cytokines, IL-6 and TNFweretested using a specific immunoassay. IL-8, IL-6, and TNFc serumlevels were significantly increased in patients with chronicrenal failure compared to their levels in normal individuals(P<0.000l, P<0.05 and P<0.000l respectively). The mostpronounced incre ment in IL-8, IL-6 and TNF serum levels wasobserved in CAPD patients (P<0.000l). CAPD patients withoutperitonitis showed relatively low levels of IL-8 or IL-6 inperitoneal dialysate effluents (PDE), whereas PDE-TNF were notdetectable in almost all patients tested. Patients with peritonitisshowed very high serum and PDE levels of IL-8, IL-6 and TNF.The clinical recovery from peritonitis was characterized bya rapid fall in IL-8, IL-6 and TNF in serum and dialysate. HDpatients showed a significant increase in serum levels of IL-8and also IL-6 and TNFcompared to normal individuals (P<0.05,P<0.05 and P<0.01 respectively). HD duration influencedserum levels of IL-8 and TNF since they were significantly higherin short-term HD patients than medium- or long-term HD patients(respectively P<0.05, P<0.00l for IL-8, and P<0.01,P<0.001 for TNF Pre-HD IL-6 levels were not influenced byHD duration. No major modification of IL-8 serum levels couldbe evinced after and before HD sessions in the short-term group,but concentrations of this cytokine were significantly higherafter HD in medium- and long-term HD patients (P<0.05, P<0.0lrespectively). In contrast, HD session did not influence IL-6and TNF levels. We conclude that the cytokine profile is perturbedin uraemia and during dialysis, and that this should be consideredas an inflammatory status.     

Intraperitoneal production of erythropoietin with continuous ambulatory peritoneal dialysis

Higher hematocrit and serum erythropoietin (EPO) levels have previously been shown in end-stage renal disease patients treated with continuous ambulatory peritoneal dialysis (CAPD) compared with hemodialysis. We investigated whether EPO was produced intraperitoneally in CAPD patients. EPO concentration was 3.5±0.3 mU/ml by radioimmunoassay in 26 samples of peritoneal dialysis effluent obtained from 15 CAPD patients. EPO was not detectable in the fresh unused dialysate. No correlation was observed between EPO levels in the serum and dialysis effluent. Peritoneal macrophages were isolated from the dialysis effluent of 9 CAPD patients after an overnight dwell. The culture supernatant obtained after 24 h of in vitro culture of a million cells yielded EPO of 3.5±0.3 mU/ml. Our study demonstrated that peritoneal macrophages from CAPD patients produce EPO on in vitro stimulation, and EPO is present in the dialysis effluent of CAPD patients.     

Lipopolysaccharide-binding protein is present in effluents of patients with Gram-negative and Gram-positive CAPD peritonitis

Background: Bacterial peritonitis is a frequent complication during treatment of end-stage renal failure by continuous ambulatory peritoneal dialysis. Local host defence mechanisms including the secretion of proinflammatory cytokines by peritoneal macrophage are of particular importance in the pathogenesis of infectious complications. LPS-binding protein (LBP) and soluble CD14 (sCD14) are serum factors known to regulate the endotoxin-induced cellular immune response. However, it is still unknown whether LBP and sCD14 are also present in the peritoneal effluent of CAPD patients. Methods: Using specific immunoassays, we examined the concentration of LBP, sCD14 and the proinflammatory cytokines TNF=&agr;, IL-1{beta} and IL-6 in the dialysis effluents of 31 patients with CAPD-associated peritonitis. Twenty patients without peritonitis served as controls. Intraperitoneal LPS concentrations were determined using the limulus amebocyte lysate assay. Results: Bacterial lipopolysaccharide could be detected in 42% of the infected dialysis effluents. In comparison to controls (0.2±0.05 &mgr;g/ml), LBP was significantly elevated in both Gram-negative/LPS-positive (1.03±0.3 &mgr;g/ml) and Gram-positive infections (0.5±0.14 &mgr;g/ml) (P<0.05). No significant differences were detected concerning the intraperitoneal sCD14 levels in the three patient groups. Levels of TNF-&agr;, IL-1{beta} and IL-6 were significantly increased in the effluents of patients with bacterial peritonitis compared to non-infected controls. Moreover the respective cytokine concentrations were significantly higher in the Gram-negative/LPS-positive compared to the gram-positive bacterial infections (P<0.01). Conclusion: Our data demonstrate that LBP is significantly elevated in the dialysis effluents of patients with CAPD-associated peritonitis caused by both Gram-negative and Gram-positive bacteria and might be used as a marker of intraperitoneal infection. Moreover, our findings support the concept that LBP enhances the effects of LPS on cytokine production by peritoneal macrophages. The function of LBP in Gram-positive infection remains to be further elucidated. Key words: CAPD-associated peritonitis; cytokines; lipopolysaccharide-binding protein; macrophages; peritoneal dialysis; soluble CD14      

Treatment of gram-positive peritonitis with two intraperitoneal doses of vancomycin in continuous ambulatory peritoneal dialysis patients

Eight patients with end-stage renal failure on continuous ambulatory peritoneal dialysis (CAPD), who developed peritonitis, received an intraperitoneal dose of vancomycin (30 mg/kg body weight) with 6 h of peritoneal dwell and then resumed their routine CAPD schedule. Vancomycin concentration in serum, peritoneal dialysate (PD) from an overnight dwell and 1, 2 and 3 h after a new exchange was measured at 48 h (in 5 patients) and 7 days (in 6 patients). Except for an occasional 1-hour peritoneal fluid sample on the 7th day, all samples had satisfactory vancomycin levels. Five of the 8 patients who had gram-positive peritonitis and 1 with 'sterile' peritonitis received another similar intraperitoneal dose of vancomycin at the 7th day. All of these patients had good therapeutic response with a negative PD culture 3 weeks after the cessation of therapy and no relapse of infection in at least 1 month of follow-up. We conclude that 2 intraperitoneal doses of vancomycin (30 mg/kg body weight) given 1 week apart with 6 h of intraperitoneal dwell is an effective and adequate treatment for gram-positive and 'sterile' peritonitis in CAPD patients.     

Inverse MCP-1/IL-8 ration in effluents of CAPD patients with peritonitis and in isolated cultured human peritoneal macrophages

An important event in intraperitoneal inflammation is the influx of leukocytes into the peritoneal cavity. Chemokines such as interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) play a major role in the recruitment of immune cells to the site of inflammation. We determined the concentrations of two members of the chemokine family, IL-8 and MCP-1, in the dialysate effluents of 18 continuous ambulatory peritoneal dialysis (CAPD) patients with peritonitis and of 18 non-infected CAPD patients by specific enzyme-linked immunosorbent assays (ELISA). Isolated peritoneal macrophages (PMs) from CAPD peritonitis patients were cultured and IL-8 and MCP-1 production was determined on protein (ELISA) and mRNA level (Northern blot) at designated timepoints over a 72-h culture period. PMs from non-infected patients served as controls. Much higher concentrations of IL-8 and MCP-1 were found in dialysate effluents of peritonitis patients than in effluents of non-infected patients: IL-8 2.39&plusnn;1.15 vs 0.05±0.01 ng/ml and MCP-1 release by cultured PMs from peritonitis patients and non-infected patients revealed significant differences: IL-8 40.3±2.2 ng/ml after 3 h and 194.2 ±34.9 ng/ml after 12 h compared to 21.02±6.15 ng/ml after 3 h and 89.64±30.28 ng/ml after 12 h, respectively; MCP-1 3.3±0.9 ng/ml after 3 h and 25.7±7.4 ng/ml after 12 h compared to 1.1±0.2 ng/ml and 1.8±0.2 ng/ml, respectively. Interestingly, the ratio of IL-8 to MCP-1 concentrations in the dialysate effluents (1:9.4) is reversed in the supernatants of cultured PMs. In the effluents and in the culture supernatants of PMs from CAPD peritonitis patients high amounts of IL-8 and MCP-1 are detectable, suggesting that PMs are an important source for these chemokines during peritonitis. Because of the inverse ratio of IL-8 and MCP-1 in the effluents and culture supernatants it can be assumed that PMs are responsible for the MCP-1 concentration to a lesser extent than for the IL-8 concentration in the effluents.