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[Decreased effectiveness of cold-induced heparin precipitation of plasma fibronectin in infection].

Plasma fibronectin (FN) is one of the major blood opsonins. The content of the glycoprotein reduces in sepsis which in turn may aggravate the course of the infection. FN is detectable in the content of cryoglobulins and cryofibrinogen. The formation of the heparin precipitate following plasma incubation in the cold in the presence of heparin is determined by FN involvement. Fibrinogen (FG) is another main component of the heparin precipitate. To determine the functional activity of plasma FN in sepsis and other pathological conditions, a study was made of the ability of FN and FG to go into the precipitate formed in blood plasma in the cold after its incubation with heparin. Unlike normal subjects in whom over 80% of FN on the average and about 20% of FG went into the heparin precipitate, in patients with hemoblastoses and aplastic anemia complicated by sepsis, less than 40% of FN on the average and about 7% of FG went into the precipitate. In some patients with sepsis, the heparin precipitate did not form. The reduction of FN ability to go into the heparin precipitate correlated with the gravity of the patients' condition. In uncomplicated hemoblastoses, cryoglobulinemia and cryofibrinogenemia and in immunocomplex pathology, the consumption of FN and FG during heparin precipitate formation did not significantly differ from the control. The data indicate that sepsis patients with blood system pathology may develop not only quantitative FN deficiency in the blood but also disorder of the functional activity of the opsonin.

Adolescent↗

Fibronectin concentration is decreased in plasma of severely ill patients with disseminated intravascular coagulation.

Plasma fibronectin (cold-insoluble globulin) is known to be cross-linked to fibrin during the final stage of blood coagulation and is probably the major nonspecific opsonin of blood. We measured the concentration of plasma fibronectin in 36 hospitalized patients (11 with malignancy, 12 with infection, 13 with other underlying diseases) with evidence of fibrin depostion and lysis. Plasma fibronectin concentration was greater than 2 S.D. below the mean of normals in 17 of the patients (p less than 0.001). Depression of fibronectin was not related to severity of disseminated intravascular coagulation, as assessed by fibrinogen concentration and the quantity of FDP in serum. Depressed plasma fibronectin concentration and the quantity of FDP in serum. Depressed plasma fibronectin concentration was an unfavorable prognostic finding, inasmuch as 12 of the 17 patients with depressed fibronectin concentrations died during hospitalization as compared to five of the 19 patients with normal fibronectin concentrations (p less than 0.02). We speculate that specific depletion of plasma fibronectin, because of codeposition with fibrin or due to increased utilization as a nonspecific opsonin, may contribute to the organ failure seen in severely ill patients.

Adult↗

[Fibronectin: an important component of bronchoalveolar defense?].

Fibronectin, an important nonimmune opsonin, was found in the bronchoalveolar lavage fluid (BAL) of 29 healthy volunteers using a sensitive radioimmunoassay. The mean FN content was 9.5 micrograms FN/mg albumin. The BAL FN was immunologically and functionally similar to plasma FN and showed opsonic activity which increased uptake of gelatin coated beads by human alveolar macrophages. Therefore, BAL contains a functionally intact FN which may serve as an opsonin for the clearance of particles and microorganisms in the terminal airways.

Bronchi↗

Humoral factors in host defense against microbial invaders.

Normal host defense against microbial invaders depends upon an intact humoral and cellular system of inflammation and specific immunity. The major weapons for killing microbes that invade tissue are phagocytic cells, however, humoral factors enable these phagocytic cells to recognize, migrate toward, efficiently ingest, and kill the invaders. Humoral factors essential for host defense include specific antimicrobial immunoglobulin antibodies and multiple proteins which comprise the complement system. The complement system is the oldest phylogenetically and the most important protein in this system is C3. When C3 is activated on the surface of the microbe, ligan is formed which provides a firm bond between organisms and phagocytic cells. This process is termed opsonization and activated C3/C3b is a highly efficient opsonin. C3b attachment stimulates efficient engulfment by phagocytic cells and most microbes are rapidly killed once they are ingested. Complement may also function as a direct bactericidal factor when associated with specific antibody on the bacterial surface. Neisseria meningitidis and Neisseria gonorrheae are susceptible to this action of complement which requires participation of the terminal proteins of the system C6, 7, 8, and 9. Patients without these terminal components, therefore, are susceptible to systemic infections with Neisseria. Septicemia from Streptococcus pneumoniae and other encapsulated microbial species occurs with greatly increased frequency in patients without spleens. Complement and other as yet unidentified humoral factors which contribute to host defense against microbial invaders appear to be deficient in patients without spleens. Antimicrobial antibodies produced in response to antigenic stimulation by microbes are also essential humoral factors. Effective levels of antimicrobial antibodies develop only after microbes have colonized their host and the immune system has been stimulated and protection is delayed. Once produced, however, specific antibodies are potent opsonins and are effective activators of complement. Highly effective and efficient host defense is present when there are circulating specific antibacterial antibodies and an intact complement system. Septicemia may occur when there is a traumatic portal or entry of an exceptionally large bacterial challenge, however, deficiency of humoral or cellular factors of host defense is frequently involved in the pathogenesis of septicemia. Measures for preventing septicemia in susceptible persons include active immunization with microbial antigens or passive therapy with specific antimicrobial antibodies.

Antibody Formation↗

Variation among cows in the ability of their blood polymorphonuclear leucocytes to kill Escherichia coli and Staphylococcus aureus.

Suspensions of polymorphonuclear leucocytes (PMN) from fresh bovine blood contained about 90 per cent neutrophils and 10 per cent eosinophils. Such suspensions had no bactericidal activity against Escherichia coli or Staphylococcus aureus, in the absence of an opsonising agent, in Hanks' balanced salt solution. Suitable opsonins were present in dilute (0.5 per cent), heat-inactivated serum or in 5 per cent whey. After incubation with PMN for two hours in the presence of the dilute serum the activity of PMN from several cows against the two organisms was highly correlated (r = 0.98) but varied significantly (P less than 0.001) among the animals from less than 2 to 30 per cent survival of the bacteria. This bactericidal activity did not alter if PMN from a single cow were incubated with sera from other animals, indicating that the critical factor was the cells and not the serum. The ranking of the animals by the bactericidal activity of their PMN was unaltered if 5 per cent whey was used as the opsonin and the leucocytes were sonicated after incubation.

Animals↗

Cooperation between Fc gamma receptor II and complement receptor type 3 during activation of platelet-activating factor release by cytokine-primed human eosinophils.

After priming with cytokines, such as granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-3, or IL-5, eosinophils are stimulated potently by opsonized particles like serum-treated zymosan (STZ), resulting in activation of the respiratory burst and production of lipid mediators, such as platelet-activating factor (PAF) and leukotriene C4 (LTC4). In the present study, the role of the opsonin receptors Fc gamma RII and CR3 during both STZ-induced activation of the respiratory burst and PAF release by human eosinophils was investigated. Inhibition studies with blocking mAbs (alpha hFc gamma RII: AT10, IV.3; alpha CR3: B2.12, 44a) showed that both Fc gamma RII and CR3 are important for STZ-induced PAF release by cytokine-primed eosinophils. In contrast, CR3 is involved in activation of the respiratory burst, whereas Fc gamma RII seems not to be important, because blocking anti-Fc gamma RII mAbs had no effect. Subsequently, experiments were performed with zymosan particles coated with IgG, iC3b, or a combination of both. IgG-coated particles poorly activated both responses in GM-CSF primed and unprimed cells. iC3b-Zymosan activated the respiratory burst as well as zymosan expressing both opsonins (IgG/iC3b-zymosan). In contrast, iC3b-zymosan induced significantly less PAF release by GM-CSF-primed eosinophils than did IgG/iC3b-zymosan, suggesting synergism between Fc gamma RII and CR3. This synergistic effect was not observed when IgG-zymosan and iC3b-zymosan were added simultaneously. Therefore, these data indicate that on human eosinophils, Fc gamma RII and CR3 act synergistically to activate PAF release, provided that their ligands are in close proximity.

Cytokines↗

[Local defense factors in the peritoneal cavity in patients on chronic peritoneal dialysis].

Examination of the risk factors for peritonitis appearance in patients on chronic peritoneal dialysis is on great importance for better understanding of its therapy and prevention. The aim of the study was to confirm the significance of some local defense parameters of peritoneal space and its role in peritonitis incidence. The methodology was: radial immunodifussion (IgG), nephelometry (C3), ingestion of the yeast particles (phagocytosis) and bioassays for cytokine determination. The level of termostable opsonines (IgG) was significantly lower in patients with high peritonitis incidence. Also, TNF, IL6 and IL1 activity was elevated or very high in 42.1%, 57.8% and 47.3% in the same group of patients but almost unmeasurable in patients with low peritonitis incidence. Phagocytosis by peritoneal macrophages was similar in all patients and the level of termolabile opsonines was critically low in the peritoneal fluid of all patients. The presented differences in local defense parameters may partly explain the susceptibility to infection in some patients being on chronic peritoneal dialysis.

Complement C3↗

Peritoneal dialysate IgG/C3 levels do not predict susceptibility to peritonitis.

OBJECTIVE: To investigate the usefulness of dialysate IgG and C3 concentrations in predicting likelihood of developing peritonitis. DESIGN: Prospective, longitudinal, and comparative study. SETTING: Single university teaching hospital dialysis unit and outpatient clinic. PATIENTS: Thirty-four uremic patients were studied (20 males, 14 females: mean age 47.2, range 20-73 years). Monthly serum and overnight dialysate (eight- to eleven-hour dwell) samples were obtained for IgG and C3 estimations over the first six months of the study, and trimonthly samples were obtained thereafter. All patients performed exchanges using standard transfer sets (Baxter system II, Baxter Healthcare Ltd., Thetford, Norfolk, U.K.), used no hypertonic fluid (3.86%) for overnight exchanges, and were followed up for a minimum of 18 months. OUTCOME MEASURES: Dialysate and serum levels of IgG and C3; peritonitis episodes. RESULTS: Forty-five episodes of peritonitis occurred in 24 patients during the study period. We examined opsonin levels in the group as a whole, and then in two subgroups of patients: those who remained peritonitis-free throughout the study, and those who did not. There were no significant differences between IgG and C3 levels in the two groups at any time point, and large interpatient and intrapatient variation in levels were seen. CONCLUSION: Dialysate levels of IgG and C3 from the overnight dwell are not helpful in predicting the risk of developing continuous ambulatory peritoneal dialysis peritonitis in individual patients. No correlation was found between opsonin levels and onset of clinical peritonitis.

Adult↗

Assessment of the functional capacity for intracellular death and phagocytosis of polymorphonuclear cells in healthy neonates.

The objective of this study was to assess the functional capacity for intracellular death (ID) and/or phagocytic index (PI) of polymorphonuclear cells of 24-h-old healthy newborns with respect to the PMN cells of adults using the same standard exogenous source of opsonins. The ID and PI techniques were standardized and 2-3 ml of blood were used. No differences were found in the percentages of ID, P, PI among the PMNs of the newborns and those of the adults: 43.95 +/- 15.70 vs. 44.56 +/- 8.43 for ID; 38.96 +/- 14.34 vs. 39.00 +/- 14.54 for P; 1.71 +/- 0.54 vs. 1.73 0.45 for PI, respectively. It was concluded that the PMNs of 24-h newborns have an ID, P, PI functionality comparable to adult PMNs; differences observed in PMN function in newborns may be due to humoral deficiencies (opsonins).

Adult↗

THE DYNAMICS OF RETICULOENDOTHELIAL BLOCKADE.

The dynamics of "reticuloendothelial blockade" were studied in living rabbits and isolated, perfused rabbit livers utilizing gelatin as a blockading agent and Au(198) stabilized in gelatin as a tracer. Employing the above experimental model, the following observations were made. (a) RES blockade was specific and dependent on the surface properties of the particle under study. (b) RES blockade was not caused by saturation of hepatic removal mechanisms. (c) RES blockade was not caused by depletion of demonstrable serum opsonins. (d) RES blockade appeared to correlate with high circulating levels of the blockading agent, per se. Thus, under the conditions employed, the term "reticuloendothelial blockade" was a misnomer. Although specificity of liver macrophage-particle interaction was evident and deserves further study, the data suggest that blockade as usually studied is a laboratory phenomenon induced by the continuing circulation of the blockading agent.

Animals↗

Effect of polymeric nanoparticle administration on the clearance activity of the mononuclear phagocyte system in mice.

We investigated the consequences of acute and subacute administration of mice with polyisobutylcyanoacrylate (PIBCA), polyisohexylcyanoacrylate (PIHCA), poly(D,L-lactic) acid (PLA), and polystyrene (PS) nanoparticles on the mononuclear phagocyte system phagocytic function. This was done by measuring the clearance rate of colloidal carbon. Single administration of PIBCA and PIHCA (but not PLA and PS) nanoparticles reduced carbon clearance in both a time- and dose-dependent fashion. Since clearance of preopsonized carbon was normal, it was assumed that PIBCA and PIHCA nanoparticles deplete opsonins specific for carbon recognition. A decrease in plasma fibronectin levels resulting from nanoparticle administration suggested its implication in their removal from blood. However, fibronectin does not seem to be responsible for PIBCA and PIHCA blockade. Phagocytic function was preserved after repeated treatment with nanoparticles, probably as a result of increased Kupffer cell phagocytic activity and the contribution of spleen macrophages. Neither toxicity nor effects due to nanoparticle hepatic accumulation were observed.

Animals↗

Opsonic activity of myeloma immunoglobulins (MM-Ig).

Patients with MM are at an increased risk for life-threatening bacterial infections, primarily by organisms that require opsonization for interaction with granulocytes. In the present study we used a neutrophil chemiluminescence (CL) assay of opsonization to explore the opsonic activity of MM-Ig for zymosan particles. Particles were treated with serum lacking in Ig to explore the contribution of Ig to zymosan opsonization. This serum was found to have 69 +/- 2.6% (X +/- SEM) of the opsonic activity of normal serum (P less than .001). Normal serum concentrations of normal IgG, MM-IgG, and MM-IgA all lacked opsonic activity for zymosan. However, when particles were treated with Nl-IgG, MM-IgG, or MM-IgA as well as complement, normal opsonic activity was generated. Increasing the concentration of the Ig to stimulate MM serum did not inhibit this opsonic activity. Thus both MM-IgG and MM-IgA can function as normal, nonspecific opsonins.

Granulocytes↗

In vivo evaluation of the effect of the size and opsonization on the hepatic extraction of liposomes in rats: an application of Oldendorf method.

In the hepatic uptake of large particles such as liposomes, a serum component called opsonin plays an important role. In this study, the 'Oldendorf method' is introduced to evaluate the hepatic extraction under the condition of single passage, which enabled examination of the effect of opsonization on liposome uptake by the intact liver. 14C-labelled liposomes and, an internal reference, 3H-H2O were injected as a bolus into portal vein. Liver uptake index (LUI) was calculated from the ratio of the extraction of 14C to that of 3H. The effect of liposome size (mean diameter of 0.8, 0.4, 0.2, and 0.05 micron) and opsonization (preincubation with fresh blood for 5 min) on liposomal hepatic uptake were investigated using this method. LUI increased with size significantly (p < 0.001), and opsonization enhanced LUI only for the large liposomes (0.8 micron). This result suggests that the critical diameter of opsonization for these liposomes lies between 0.4 and 0.8 micron.

Animals↗

Reticulo-endothelial function in obstructive jaundice.

A study of reticulo-endothelial function was performed in 30 patients with obstructive jaundice. Reticulo-endothelial phagocytic function, measured by the clearance of intravenous human micro-aggregated albumin, was significantly depressed in jaundiced patients compared with non-jaundiced controls (P less than 0.001). There was a significant correlation (P less than 0.001) between phagocytic function and plasma bilirubin level but not with transaminase or bile salt level. Phagocytic function was not related to the presence of malignancy, but was markedly reduced in patients with cholangitis. There was no reduction in hepatic sinusoidal blood flow, opsonin levels (fibronectin, IgG, complement) or serum opsonic activity to account for the reticulo-endothelial dysfunction.

Aged↗

Markers of macrophage heterogeneity. I. Studies of macrophages from various organs of normal mice.

A unique subpopulation of macrophages (M phi) was identified among the spleen and bone marrow M phi of normal mice. After 24 h of culture, approximately 2.5% of the adherent cells cluster into "foci" of 10-30 cells. On the basis of their phagocytic and morphologic characteristics, these focus-forming M phi (FF-M phi) appeared to be highly activated. Uncoated sheep erythrocytes (E) were ingested by FF-m phi indicating that opsonization was not a prerequisite for phagocytosis. However, IgM-coated E (EIgM) were more readily phagocytosed by FF-M phi than were E suggesting that IgM is recognized as an effective opsonin by these cells. EIgM and E coated with IgM and complement (C) (EIgMC) were ingested by approximately the same percentage of FF-M phi; thus, if these cells possess complement receptors in addition to structures which bind EIgM, the C receptors do not enhance the ability of FF-M phi to ingest opsonized particles. The non-focus-forming M phi, e.g. individual M phi (I-M phi), in the spleen and bone marrow can, themselves, be divided into various subpopulations distinguished by their ability to bind and ingest E, EIgM ana EIgMC. These may represent various subpopulations of M phi or M phi at various stages of activation or differentiation. While spleen and bone marrow M phi contained FF-M phi and I-M phi which vary in their ability to ingest E, EIgM and EIgMC, theM phi of the peritoneum and blood of normal mice were far more homogeneous. Peritoneal and blood M phi did not form foci, ad did not ingest E or EIgM in significant amounts although a small percentage were able to ingest EIgMC. These data suggest that the population of M phi in the spleen and bone marrow are far more heterogenous than those found in the peritoneum or blood and that binding and phagocytosis of various coated and uncoated erythrocytes can be studied to elucidate this heterogeneity.

Animals↗

The role of extracellular matrix proteins in the control of phagocytosis.

The phagocytic function of human-peripheral-blood-derived mononuclear and polymorphonuclear leukocytes can be regulated by external stimuli. In particular, the extracellular matrix (ECM) proteins fibronectin and laminin and serum amyloid P component can increase the phagocytic activity of these cells. Phagocytosis enhancement by the ECM requires two distinct signals to the ingesting cell. First, a ligand for interaction of the target particle with phagocytic cells is required. Generally this occurs because of opsonins such as antibody or complement on the phagocytic target and is independent of the ECM proteins. The phagocytosis-enhancing effect of the connective tissue proteins also requires direct interaction of these proteins with phagocytic cells and occurs through cell surface receptors for these molecules. In the case of neutrophils, sensitivity to the phagocytosis-enhancing effects of connective tissue proteins requires prior stimulation with the chemotactic peptides C5a or f-met-leu-phe. The present state or knowledge of the mechanism of phagocytosis enhancement by ECM proteins and the implications of the phenomenon for host defense are discussed.

Animals↗

Involvement of complement receptor 3 (CR3) and scavenger receptor in macrophage responses to wear debris.

The ability of prosthetic wear debris to induce pro-inflammatory responses in macrophages is widely appreciated, but little is known about the molecular mechanisms involved in particle recognition. Specifically, the nature of the cell surface receptors that interact with wear debris is poorly understood. Elucidating the identities of these receptors and how they interact with different types of wear debris are critical to understanding how wear debris initiates periprosthetic osteolysis. We examined the involvement of opsonization, complement receptor 3 (CR3), and scavenger receptor A (SRA), in responses to polymethylmethacrylate (PMMA) and titanium wear particles. Serum dependence of pro-inflammatory responses to PMMA and titanium was tested, and serum proteins that adhered to these two types of particles were identified. Several serum proteins, including known opsonins such as C3bi and fibronectin, adhered to PMMA but not titanium, and serum was required for pro-inflammatory signaling induced by PMMA, but not by titanium. Phagocytosis of PMMA and titanium by macrophages was demonstrated by flow cytometry. Blocking CR3 specifically inhibited phagocytosis of PMMA by macrophages, whereas blocking SRA specifically inhibited titanium uptake. Direct involvement of CR3 and SRA in cell-particle interaction was assessed by expression of these receptors in nonphagocytic HEK293 cells. CR3 specifically induced cell binding to PMMA particles and adhesion to PMMA-coated plates, while SRA specifically induced binding to titanium particles and adhesion to titanium-coated plates. Taken together, these results suggest involvement of opsonization, complement, and integrin receptors, including CR3 and fibronectin receptors, in PMMA action, and an involvement of scavenger receptors in responses to titanium.

Fibronectins↗