lactoferrin and Uremia

Multiple dysfunctions of polymorphonuclear leukocytes (PMNLs) contribute significantly to the increased morbidity and mortality among patients with end-stage renal disease. In the present study, we measured the PMNL content of beta(2)-microglobulin (beta(2)m) and lactoferrin in different states of renal insufficiency and after kidney transplantation. PMNLs were lysed ultrasonically and, after centrifugation, both proteins were assayed in the supernatant by enzyme-linked immunosorbent assay technique. Despite marked differences in plasma beta(2)m levels, no significant difference in PMNL content of beta(2)m and lactoferrin could be shown among the groups analyzed. There was also no correlation between plasma beta(2)m level and PMNL beta(2)m content. In control subjects, as well as in renal allograft recipients with a well-functioning graft, PMNL beta(2)m level correlated positively with PMNL lactoferrin level (pooled data, r = 0.55; P < 0.001; n = 55). Both proteins are considered to colocalize in peroxidase-negative PMNL granules. However, no correlation was found in the azotemic and uremic patient groups. Standard immunofluorescence staining of control PMNLs showed a cytoplasmic granular distribution of both granule proteins. However, in PMNLs of uremic patients, lactoferrin shifted to a perinuclear localization. PMNLs obtained from uremic individuals failed to elicit an increase in lactoferrin release after stimulation with the chemotactic peptide f-Met-Leu-Phe compared with PMNLs obtained from healthy volunteers. These data indicate abnormalities in uremic patients of PMNL granule lactoferrin content and release that are reversible after successful renal transplantation.

Topics: Adult; beta 2-Microglobulin; Cell Degranulation; Cell Nucleus; Chemotactic Factors; Cytoplasmic Granules; Enzyme-Linked Immunosorbent Assay; Fluorescent Antibody Technique, Direct; Humans; Kidney Failure, Chronic; Kidney Transplantation; Lactoferrin; Middle Aged; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Peritoneal Dialysis, Continuous Ambulatory; Renal Dialysis; Transplantation, Homologous; Uremia

Recently, we developed a new method to measure the resting level of superoxide anion in whole blood using an ultrasensitive chemiluminescence analyzer and lucigenin amplification. The advantage of this method is that the assay system can be performed in the absence of leukocyte isolation and stimulant administration. In this study, we applied this method to measure the blood resting levels of superoxide anion in 104 uremic patients on chronic hemodialysis (CHD) and 98 sex- and age-matched healthy controls to clarify the influence of HD on blood levels of superoxide anion. Simultaneously, the plasma levels of copper, zinc superoxide dismutase (Cu,Zn-SOD), glutathion peroxidase (GPX), myeloperoxidase (MPO) and lactoferrin (Lacto-F) were measured. The results showed that the basal blood levels of superoxide anion, Cu,Zn-SOD, and MPO in CHD patients were significantly greater than those of healthy controls. However, there was no difference in the basal plasma levels of Lacto-F and GPX between CHD patients and healthy controls. One session of HD further increased the blood levels of superoxide anion, MPO, Lacto-F and Cu,Zn-SOD but not GPX. These results suggest that the blood levels of superoxide anion are higher in CHD patients and further increase after one session of HD. This mechanism should be studied further.

Topics: Acridines; Adult; Aged; Alcohol Drinking; Female; Glutathione Peroxidase; Humans; Lactoferrin; Luminescent Measurements; Male; Middle Aged; Peroxidase; Renal Dialysis; Rest; Smoking; Superoxide Dismutase; Superoxides; Uremia

Topics: Adult; Erythropoietin; Ferritins; Humans; Iron; Lactoferrin; Middle Aged; Recombinant Proteins; Renal Dialysis; Uremia

Removal of beta 2-microglobulin has become a major objective of dialysis therapy. The present study was performed to evaluate both compatibility and elimination capacity for beta 2-microglobulin of a newly developed high-flux polyamide membrane (Polyflux 130) during hemodialysis. The degree of leukopenia was moderate (-22%) and comparable with Polysulfone 600 (-25%). C3a desarg generation had a tendency to be lower with the Polyflux 130 membrane, and C5a desarg formation was identical with both types of membranes. As for degranulation of polymorphonuclear leukocytes, plasma elastase levels increased by 209% with Polyflux 130 and by 160% with Polysulfone 600 membranes. Likewise, plasma lactoferrin values rose during hemodialysis by 233% (Polyflux 130) and 160% (Polysulfone 600). The differences between membranes, however, were statistically not significant. There was a sharp drop in the serum levels of beta 2-microglobulin during dialysis with both membranes (Polyflux 130: -46%; Polysulfone 600: -48%). Accordingly, sieving coefficients were calculated to be 0.77 +/- 0.06 for Polyflux 130 and 0.80 +/- 0.06 for the Polysulfone 600 membrane. Both membranes were capable to remove large quantities of beta 2-microglobulin, amounting to 235 +/- 11 and 250 +/- 10 mg/4 h of dialysis for Polyflux 130 and Polysulfone 600, respectively.

Topics: Adult; Amyloidosis; beta 2-Microglobulin; Cellulose; Complement Activation; Evaluation Studies as Topic; Humans; Lactoferrin; Leukopenia; Membranes, Artificial; Middle Aged; Nylons; Pancreatic Elastase; Polymers; Renal Dialysis; Sulfones; Uremia

Plasma levels of granulocyte elastase in complex with alpha 1-proteinase inhibitor (E-alpha 1-PI) increased during hemodialysis from 110 +/- 10 to 506 +/- 66 micrograms/L using dialyzers made of polyamide, from 95 +/- 2.2 to 211 +/- 54 micrograms/ml with hemophane and from 114 +/- 10 to 203 +/- 25 using dialyzers made of polysulfone. Plasma lactoferrin values were also significantly higher during hemodialysis with polyamide (772 +/- 110 micrograms/L) compared with hemophane (268 +/- 2.2) and the polysulfone (278 +/- 31 micrograms/L) dialyzer. After dialysis each dialyzer was rinsed. We found the lowest concentration of lactoferrin (902 +/- 254 micrograms/L) and E-alpha 1-PI (739 +/- 162 micrograms/L) after rinsing polysulfone dialyzers, whereas the highest concentrations were observed after rinsing the polyamide dialyzer (lactoferrin: 2,426 +/- 314; E-alpha 1-PI: 1,134 +/- 144 micrograms/L). Hemodialysis with polysulfone dialyzers caused significantly lower plasma levels of beta-2-microglobulin compared with polyamide or hemophane membranes despite significantly lower levels in the rinsing solutions. Our data indicate that low plasma levels of main granulocyte component observed with polysulfone and hemophane dialyzers are not the result of higher membrane adsorption of E-alpha 1-PI and lactoferrin. These main granulocyte components are not related to beta-2-microglobulin levels of both plasma and rinsing solutions.

Topics: Aged; Aged, 80 and over; alpha 1-Antitrypsin; beta 2-Microglobulin; Blood Proteins; Humans; Kidneys, Artificial; Lactoferrin; Lactoglobulins; Membranes, Artificial; Middle Aged; Protease Inhibitors; Renal Dialysis; Uremia

The effect of the calcium channel blockers nifedipine (9 and 18 micrograms kg-1 h-1), diltiazem (100 and 200 micrograms kg-1 h-1) and verapamil (19 micrograms kg-1 h-1) continuously infused during haemodialysis on granulocyte and complement activation was investigated. Plasma levels of lactoferrin, elastase in complex with alpha 1-proteinase inhibitor (E-alpha 1PI) and C3a were measured in patients dialysed with dialysers made of cuprophane, polymethylmethacrylate (PMMA) and polyacrylonitrile (PAN). Calcium channel blockers caused no change of blood pressure during haemodialysis in all patients. There was no effect of nifedipine, diltiazem or verapamil on plasma lactoferrin, E-alpha 1PI or C3a levels in patients dialysed with cuprophane. However, plasma lactoferrin and E-alpha 1PI values were significantly reduced by all calcium channel blockers in patients dialysed with PMMA, and also by nifedipine and verapamil in patients dialysed with PAN. Our data indicate that calcium channel blockers inhibit granulocyte activation occurring in dialysers with very little anaphylatoxin formation. These drugs, however, are ineffective in patients dialysed with cuprophane where complement activation takes place. Therefore, granulocyte activation during haemodialysis in the absence of complement activation seems to be mediated by calcium ions.

Topics: Aged; alpha 1-Antitrypsin; Blood Proteins; Calcium Channel Blockers; Complement Activation; Female; Granulocytes; Humans; Kidneys, Artificial; Lactoferrin; Leukopenia; Male; Middle Aged; Pancreatic Elastase; Renal Dialysis; Uremia

Plasma levels of granulocyte lactoferrin, granulocyte myeloperoxidase and granulocyte elastase in complex with alpha 1-proteinase inhibitor (E-alpha 1PI) were investigated in regular hemodialysis patients dialyzed with hollow-fiber dialyzers made from polycarbonate (FD 100) or cuprophan (GFS 120 H). Plasma levels of all these main granulocyte components increased significantly during hemodialysis. E-alpha 1PI levels were significantly higher in patients dialyzed with the polycarbonate compared with the cuprophan membrane, whereas the increases of myeloperoxidase and lactoferrin were not different for the two dialyzers. On the other hand, plasma C3a levels were higher in patients dialyzed with the cuprophan compared with the polycarbonate dialyzer. Therefore, granulocyte activation during hemodialysis does not necessarily need complement activation.

Topics: alpha 1-Antitrypsin; Blood Proteins; Cellulose; Complement C3; Complement C3a; Dental Cements; Female; Granulocytes; Humans; Kidneys, Artificial; Lactoferrin; Male; Membranes, Artificial; Middle Aged; Pancreatic Elastase; Peroxidase; Polycarboxylate Cement; Renal Dialysis; Uremia

Recently, a new type of modified cellulosic membrane (Hemophan) has been made available for the first clinical trials. In contrast to cellulose acetate membranes only about 5% of the hydroxyl groups of cellulose are substituted by tertiary amino groups. Objective of the present study was to assess the compatibility performance of this new membrane material. Therefore, 10 patients were dialyzed consecutively with membranes made of regenerated cellulose (Cuprophan) and modified cellulose. The modified cellulosic membrane showed less leukopenia. The generation of C3adesarg was reduced throughout the dialysis session. Both, the release of lactoferrin and elastase from granulocytes was diminished with the modified cellulosic membrane. All these differences reached the level of significance. Therefore, it seems that with Hemophan a more biocompatible type of cellulosic membrane has been introduced into hemodialysis treatment.

Topics: Biocompatible Materials; Cellulose; Complement Activation; Granulocytes; Humans; Lactoferrin; Leukopenia; Membranes, Artificial; Middle Aged; Pancreatic Elastase; Renal Dialysis; Uremia

Plasma C3a and C5a levels as well as plasma levels of granulocyte lactoferrin, granulocyte myeloperoxidase and granulocyte elastase in complex with alpha 1-proteinase inhibitor (E-alpha 1PI) were investigated in 10 patients (52.7 +/- 5.9 years) undergoing maintenance hemodialysis (39.4 +/- 12.4 months) with hollow fiber dialyzers made from polymethylmethacrylate. Plasma levels of lactoferrin increased from 166.5 +/- 28.5 to 712.5 +/- 165.9 ng/ml, myeloperoxidase from 59.0 +/- 15.3 to 210.5 +/- 33.9 ng/ml and E-alpha 1PI from 114.2 +/- 18.1 to 681.8 +/- 102.6 ng/ml during dialysis. In contrast, plasma C3a levels rose from 179.8 +/- 33.6 to maximal 276.2 +/- 45.4 ng/ml and C5a from 55.7 +/- 8.1 to maximal 101.1 +/- 14.8 ng/ml. Our data indicate that degranulation of granulocytes occurs during dialysis despite only little complement activation and mild initial granulocytopenia.

Topics: alpha 1-Antitrypsin; Biocompatible Materials; Blood Proteins; Complement Activation; Complement C3; Complement C3a; Complement C5; Complement C5a; Female; Granulocytes; Humans; Kidneys, Artificial; Lactoferrin; Lactoglobulins; Male; Membranes, Artificial; Methylmethacrylates; Middle Aged; Pancreatic Elastase; Peroxidase; Protease Inhibitors; Renal Dialysis; Uremia

The present study was devised to investigate, whether reduction of membrane surface area would contribute to enhanced dialyzer membrane compatibility. Therefore, 10 hemodialysis patients were dialyzed with three different cuprophan dialyzers, displaying membrane surface areas of 0.9 m2, 1.2 m2, and 1.8 m2. As an index of biocompatibility release of granulocyte elastase and secretion of granulocyte lactoferrin were determined by enzyme-linked immunoassays. With the three cuprophan membranes, marked leukopenia occurred 15 min after the start of hemodialysis, without significant distinction between the different membranes. On the other hand, the release of leukocyte elastase was strictly dependent on the membrane surface area. Thus, at the end of dialysis, plasma levels of elastase were as follows: 0.9 m2: 255 +/- 51; 1.2 m2: 356 +/- 65; 1.8 m2: 471 +/- 56 ng/ml. The magnitude of secretion of leukocytic lactoferrin was also dependent on membrane surface area. Therefore, the increment during dialysis was 300% with the 1.2 m2 membrane compared to 650% using a 1.8 m2 dialyzer. Based upon these data, we concluded that, using measurements of granulocyte degranulation as an index of biocompatibility, reduction of membrane area resulted in a marked improvement of membrane compatibility.

Topics: Adult; Aged; Biocompatible Materials; Cellulose; Humans; Kidney Failure, Chronic; Kidneys, Artificial; Lactoferrin; Leukocyte Count; Membranes, Artificial; Middle Aged; Pancreatic Elastase; Uremia

Transient reduction in circulating polymorphonuclear granulocytes and eosinophils were observed early in hemodialysis. About a threefold increase in serum-lactoferrin occurred 2 h from the start of hemodialysis. Increments of the serum levels of eosinophil cationic protein (ECP) were observed as early as 1 h after initiation of hemodialysis, reaching maximum levels (about a fourfold increase from initial levels) 1 h later. When fresh blood was circulated through a dialyzer without having a patient in the circuit considerable increases of lactoferrin and ECP were also found. The intracellular contents of lactoferrin and ECP in granulocytes isolated from peripheral blood were unaffected throughout the dialysis period. Sera obtained at different times during dialysis induced no release of granular proteins from isolated granulocytes in vitro. The raised serum concentrations of lactoferrin and ECP during dialysis suggest that a local degranulation of neutrophils and eosinophils may take place probably in the dialyzer.

Topics: Adult; Blood Proteins; Eosinophil Granule Proteins; Eosinophils; Humans; Kidneys, Artificial; Lactoferrin; Lactoglobulins; Muramidase; Neutrophils; Renal Dialysis; Ribonucleases; Uremia