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Circulating immune complexes and renal function in canine leishmaniasis.

A third component of complement (C3) capture enzyme-linked immunosorbent assay was used to determine the concentration of IgG circulating immune complexes (CIC) in 91 dogs with naturally acquired leishmania infection and in a control group of 24 healthy dogs. Results were expressed as a percentage of a reference standard. Mean concentrations of CIC were significantly (P < 0.001) higher in leishmania-infected dogs (228.725 +/- 14.283%) than in controls (74.542 +/- 12.614%). An increase in CIC concentration was found in 57.1% of the leishmania-infected dogs. No significant differences could be recorded in CIC levels between males and females in either group. Infected dogs showing hypercreatininemia rendered a statistically significant (P < 0.030) higher serum CIC concentration than sick dogs with normal creatininemia. When hypercreatininemia (> or = 1.30 mg/dl) was used as an indicator for CIC increase, the positive predictive value obtained was 0.9 indicating that renal function impairment was associated with high serum CIC concentration in 90% of the infected dogs.

Animals↗

Levels of complement factor C3 and its activated product, C3a, in operatively salvaged blood.

In intra-operative blood salvage the collected blood is exposed to traumatized tissue and a synthetic circuit. Because of this exposure it was expected that the complement system, which is a contact system present in plasma, would be activated in salvaged blood. To determine whether this occurs, the plasma levels of the complement factor C3 and its activated fragment, C3a, were measured in intra-operative salvaged blood before and after washing. Samples were obtained from patients undergoing aortic surgery where intra-operative salvage was used. In the unwashed salvaged blood, the level of C3 fell (mean C3: 0.33 g/L) and the level of C3a increased (mean C3a: 1994 ng/mL) compared with the patient circulating levels of C3 and C3a (mean C3: 0.70 g/L, mean C3a: 855 ng/mL) respectively. Washing of the collected blood reduced the C3a level (mean: 346 ng/mL) to the patient's level and reduced C3 to the lower detection limit of the test (less than 0.2 g/L). The raised level of C3a and the reduced level of C3 confirm that the complement system is activated and imply that other complement factors are also activated.

Blood Loss, Surgical↗

Unusual variants of pemphigoid: from pruritus to pemphigoid nodularis.

We report three patients with pemphigoid nodularis. Patients were females aged 76, 71 and 50 years, and all had features of bullous pemphigoid together with prurigo-like lesions at some stage of their illness. In two cases, nodular lesions preceded the onset of blistering by some months. Blisters arose on normal skin and in one patients also at sites of prurigo lesions. Routine histology of bullous lesions revealed the presence of subepidermal blisters. Electron microscopy performed in two cases confirmed the level of split to be through the lamina lucida. Direct immunofluorescence in all cases was positive, with linear basement membrane zone deposition of IgG and C3. Circulating IgG anti-basement membrane antibody was also detected in all patients, and in two, immunoblotting revealed a single antigen of 220 kD.

Aged↗

Serum clusterin and vitronectin in alcoholic cirrhosis.

Clusterin and vitronectin are multifunctional regulatory proteins which both serve as complement lysis inhibitors. Previous data have strongly suggested that serum vitronectin is mainly produced in the liver, whereas the biosynthetic origin for serum clusterin has not been determined. In the present study we aimed to determine the role of the liver in producing these proteins and to evaluate the proteins as possible markers of liver failure. We therefore quantified clusterin and vitronectin in serum from patients suffering from alcoholic liver cirrhosis (n = 83), and in serum-free culture supernatants from the hepatoma cell line HepG2. The median clusterin concentration was 0.20 g/l in cirrhosis and 0.37 g/l in the controls, whereas corresponding vitronectin values were 0.19 and 0.26 g/l, respectively. The concentration of both proteins showed significant correlation (p < 0.0001) with disease severity and with established plasma markers of hepatic synthetic function, such as albumin and prothrombin complex. The clusterin level, but not the vitronectin level, correlated with survival (p = 0.005). The rates of synthesis of clusterin, vitronectin and C3 from HepG2 cells were 0.02, 0.21 and 1.9 micrograms/10(6) cells/24 h, respectively. From the present data we conclude that clusterin (as vitronectin and C3) is mainly produced in the liver and may be a useful marker in the evaluation of severity of liver disease and prognosis of patients with alcoholic cirrhosis.

Adult↗

Vascular events in placentae and organ allografts.

This investigation explores the transplantation analogy of placentae with allografted human organs. Biopsies of cardiac and renal allografts and placentae were studied immunocytochemically with antibodies to components of the immunological, coagulational, anticoagulational, and fibrinolytic systems. Cellular rejection of cardiac and renal allografts was identified by infiltrating lymphocytes and macrophages. This was accompanied by vascular damage characterized by loss of endothelial anticoagulant pathways, vascular deposits of fibrin, and depletion of arterial tissue plasminogen activator (tPA). Failing allografts, including placentae from abnormal pregnancies, demonstrated coagulation/fibrinolytic changes consistent with vascular rejection, regardless of the presence of cellular infiltrates. An IgM autoantibody to allogeneic endothelium was associated with vascular protection. Its presence in cardiac and renal transplant biopsies was associated with an absence of fibrin deposits, and its absence was associated with vascular damage. Atherosclerosis commonly was identified in allograft biopsies (including placentae from abnormal pregnancies). These changes were shown in serial biopsies of transplanted hearts to be preceded by or associated with diminished IgM autoantibody, impaired endothelial anticoagulant pathways, vascular fibrin deposits, and depleted arterial reactivity for tPA. These results indicate the transplantation analogy of pregnancy should be viewed more specifically as vascular smooth muscle cell and endothelial responses to as yet unidentified microenvironmental stimuli.

Arteriosclerosis↗

A reference procedure to study chemiluminescence induced in polymorphonuclear leukocytes by Neisseria meningitidis.

Luminol-enhanced chemiluminescence (CL) was used to study the ability of various strains of Neisseria meningitidis (MC) to induce oxidative metabolism of polymorphonuclear leukocytes (PMNL); an indirect measure of phagocytic activity. To circumvent variations related to different PMNL donors, a MC serogroup X strain was used as a control for indexing the CL responses induced by other MC strains. This procedure, with pooled serum from healthy blood donors to standardize opsonising conditions, gave reproducible and comparable results, irrespective of PMNL donors. Under these conditions, there was a highly significant difference between pathogenic and non-pathogenic MC strains as regards their ability to induce CL responses (p less than 0.001). The results indicated that the differences were due partly to opsonizing antibodies, partly to other differences related to pathogenicity of tested MC strains. These differences in leukocyte/MC interaction were also confirmed by phagocytic-killing experiments. The index procedure of CL measurements may be a suitable method to study the appearance of natural immunity to MC disease, as well as the pathogenicity of particular MC strains.

Antibodies, Bacterial↗

Immunoelectron microscopy on epoxy sections without deplasticizing to detect glomerular immunoglobulin and complement deposits in renal diseases.

Twenty renal biopsies were studied by immunoelectron microscopy (IEM) after embedding in epoxy resin. Immunogold labeling for immunoglobulins and complement C3 was performed on the epoxy sections, which were not subjected to any kind of etching or deplasticizing prior to the immunolabeling. The concentration of accelerator, DMP-30 (Tri (Dimethyl Amino Methyl) Phenol), was increased in the infiltration and embedding steps far beyond the values normally used to make immunolabeling of these antigens possible on epoxy sections. The sections were stained with tannic acid accompanied by uranyl acetate and lead citrate. Immunofluorescence (IF) for light microscopy was carried out on frozen sections of parallel tissue samples. Some cases with IgA-nephritis demonstrated a higher sensitivity for IEM than IF, in the sense that smaller amounts of antigen were detectable with IEM. Ultrastructural preservation with this method was approximately the same as that usually seen on epoxy-embedded material. By combining excellent immunolabeling with nearly optimal ultrastructural morphology in one procedure, this method is useful particularly in situations where the material available is limited, such as in studies of renal biopsies. As far as we know, this is the first time that immunoglobulins have been satisfactorily immunolabeled on epoxy sections without etching or deplasticizing.

Complement C3c↗

Immunopathogenesis of canine aural haematoma.

Data are presented from 15 dogs with aural haematoma. The series included six Labrador retrievers and four golden retrievers and the mean age was 8.0 +/- 3.02 years. Five dogs had evidence of pruritic skin disease and five further cases had other concurrent disease. Haematology and serum biochemistry were normal in 12 and 13 of the 15 dogs, respectively. All dogs were Coombs' negative and serum antinuclear antibody had negative or low titres in all the 11 cases tested. Histopathological examination of biopsies from the affected ears revealed variable degrees of erosion of auricular cartilage with fibrovascular granulation tissue filling the cartilage defects. There was minimal perichondral inflammation. The biopsies were studied by immunohistochemistry for deposition of immunoglobulin G (IgG), immunoglobulin M (IgM) and complement C3. In one dog there was basement membrane zone deposition of IgG and in another there was focal interepithelial deposition of both IgG and IgM. The findings of this study do not support an autoimmune pathogenesis for canine aural haematoma, but suggest that an early immunological event may underlie the observed cartilage erosion.

Animals↗

Opsonic activity of intravenous immunoglobulin preparations against Staphylococcus epidermidis.

Peritoneal dialysis effluent collected from patients undergoing continuous ambulatory peritoneal dialysis had no opsonic activity against Staphylococcus epidermidis and contained low concentrations of IgG and C3 (roughly equal to those found in 0.5% normal human serum). An intravenous immunoglobulin preparation that showed no opsonic activity against the same organism, on its own or when added to balanced salt solution or peritoneal dialysis fluid, showed good activity when combined with peritoneal dialysis effluent. This was probably due to the presence of low concentrations of C3 in the effluent as prior heat inactivation at 56 degrees C for 30 minutes eliminated any opsonic activity in the immunoglobulin-dialysis effluent mixture. Examination of a range of immunoglobulin preparations showed that their opsonic activity for S epidermidis in the absence of complement varied considerably. Luminol dependent chemiluminescence was unsatisfactory as a method for detecting complement independent immunoglobulin mediated opsonisation. Intravenous immunoglobulin preparations may be useful in boosting the peritoneal defences of those patients undergoing continuous ambulatory peritoneal dialysis (CAPD) who suffer from repeated intraperitoneal infections.

Complement C3↗

Heparin reversal by protamine in humans--complement, prostaglandins, blood cells, and hemodynamics.

Fourteen noncardiac surgical patients received heparin (10,000 IU), which was neutralized by 100 mg protamine injected within 2 min during steady-state anesthesia. After protamine application, plasma complement C3a, thromboxane B2 (TxB2), prostaglandin F2 alpha (PGF2 alpha) and KH2PGF2 alpha increased significantly, whereas prostacyclin (6-keto-PGF2 alpha) levels did not change. This mediator response was associated with transient leukopenia and thrombocytopenia. Arterial pressure, pulmonary arterial pressure, and transpulmonary pressure gradient increased significantly. Heart rate, cardiac output, pulmonary capillary wedge pressure, and arterial PO2 remained constant. Positive correlations of plasma C3a were observed with pulmonary leukosequestration and plasma TxB2. Inverse correlations of C3a were noted with the counts of leukocytes and of platelets. A positive correlation was found between TxB2 and pulmonary arterial pressure. Our results indicate that marked activation of the complement system and the cyclooxygenase pathway is common after heparin reversal by protamine in anesthetized patients. This is in contrast to previous human studies performed after cardiopulmonary bypass but agrees well with results obtained in animal experiments. The mediator response in our patients, however, was not accompanied by hemodynamic instability, suggesting appropriate compensatory mechanisms.

Aged↗

The biocompatibility of silicone, fluorosilicone and perfluorocarbon liquids as vitreous tamponades. an ultrastructural and immunohistochemical study.

PURPOSE: To investigate the interactions occurring at the interface between some currently used vitreous tamponading substances and the ocular tissues, in an early follow-up after surgery. METHODS: Adult albino rabbit eyes underwent vitrectomy and were injected intravitreally with silicone oils at 1,000 or 3,000 cSt, fluorosilicone oil and perfluorodecalin. Different morphological techniques (light microscopy, scanning and electron microscopy, immunohistochemistry) were applied. RESULTS: All the tested materials, although non-toxic, penetrate the ocular tissues also at the anterior segment level, and in the long-term follow-up this can yield functional impairment. No massive inflammation has been detected in the zones in contact with the materials, but immunoglobulins and complement fractions are anyway present in the stroma of various tissues and around the droplets of emulsified materials, suggesting a local immune reaction. CONCLUSIONS: Data from this study confirmed that a permanent vitreous substitute, showing a perfect biocompatibility, still does not exist. The indication to remove the tamponading substance within 2 months after the surgery emerges from this investigation, confirming previous studies. Despite some negative features, silicone oil still appears the most biocompatible material for vitreous replacement.

Animals↗

Etiopathogenesis of otosclerosis.

Otosclerosis is a frequent cause of nonsyndromic hearing loss which affects exclusively the human temporal bone. Various etiopathogenetic hypotheses have been proposed. The major hypotheses considered are genetic factors, immunologic factors and viral infection. Since the familial incidence of otosclerosis is known a recent genetic analysis has given evidence of three otosclerosis genes (OTSC1-3). Mutations in the collagen gene COL1A1 have been found in one large family with several cases of otosclerosis. Concerning an immunologic etiopathogenetic process, the presence of serum antibodies against collagen II and IX in patients with otosclerosis confirms the hypothesis of a collagen autoimmune mechanism. Finally as a possible cause of this chronic inflammatory disease morphologic and biochemical investigations have revealed a measles virus association. In conclusion, various etiopathogenetic factors may contribute to the genesis of otosclerosis.

Bone Resorption↗

C3 variants in Japanese.

By high-voltage agarose gel electrophoresis, seven phenotypes of C3 were found in Japanese. The allele frequencies for C3*S, C3*S025, C3*S02, C3*F, C3*F06, C3*F065, and C3*F08 were 0.9943, 0.0003, 0.0003, 0.0006, 0.0003, 0.0021, and 0.0021, respectively. CH50, C3/C3c protein concentrations, and C3 hemolytic activities in fresh sera with variant C3 phenotypes were within the normal ranges.

Adult↗

GC and C3 serum groups in ulcerative colitis.

The phenotypes and allele frequencies of two serum protein systems (GC and C3) were studied in 91 consecutive patients with ulcerative colitis and compared with healthy controls. No significant differences were found as far as GC was concerned. However, significant differences were observed in C3: the C3*F allele and C3FS phenotype were more frequent in patients than in controls.

Alleles↗

Effects of repeated sampling on the properties of the cervical mucus during one day at midcycle.

Midcycle cervical secretions were collected every hour from 08.00 to 12.00 h in 12 normal women. There was a significant decrease in gram wet weight of mucus in the first four samples followed by an increase in the fifth specimen (trend analysis). Spinnbarkeit decreased significantly during repeated sample collection. Albumin concentration in cervical secretion increased significantly during repeated sample collection, but there was no statistically significant difference in the total amount of albumin recovered during repeated sample collection. The possibility of a hormonal regulation of the water content in the cervical mucus is discussed.

Adult↗

Changes in immunological recovery in patients who received post-transplant G-CSF or GM-CSF after autologous peripheral blood stem cell transplantation (PBSCT).

In this prospective study, the effects of granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) on immunological reconstitution after autologous peripheral blood stem cell transplantation (PBSCT) were investigated for 6 months. Thirty-five patients received G-CSF 5 microg/kg per day and 26 patients received GM-CSF SC 5 microg/kg per day from day 1 to leukocyte engraftment (>1000 per mm3). Peripheral blood samples were obtained on 14, 28, 100, and 180 days after transplantation for immunological evaluation. CD3+, CD4+, CD8+, CD19+, and CD56+ cells were analysed by flow cytometry. Immunoglobulin levels (IgG, IgA, and IgM) and complement levels (C3c and C4) were measured by nephelometry. Both G-CSF and GM-CSF groups were comparable with respect to age, sex, the period from diagnosis to transplantation, total nucleated cells infused, the number of CD34+ cells, conditioning regimens (TBI and non-TBI), and post-transplant infection. CD3+ and CD8+ cells on day 14 following autologous PBSCT + G-CSF were significantly higher than following autologous PBSCT + GM-CSF (p = 0.008 and p = 0.021, respectively). The number of CD4 cells and the CD4/CD8 ratio were not different at several time points between the two groups. CD19+, CD56+ cells and immunoglobulin levels showed a faster recovery pattern in the autologous PBSCT + G-CSF group. The effect of G-CSF on immune reconstitution after autologous PBSCT is more prominent than that of GM-CSF. The possible role of haematopoietic growth factor on immune recovery and its clinical importance should be investigated in further studies.

Adult↗

Impact of endograft material on the inflammatory response after elective endovascular abdominal aortic aneurysm repair.

The purpose of this paper is to examine the impact of endograft material on the inflammatory response after elective endovascular abdominal aortic aneurysm repair. Consecutive patients (n = 22, all men, 53 to 82 years old) were divided into 2 groups according to the graft material used: In group A (n = 12) the endovascular device was made of polyester and in group B (n = 10) the device was made of expanded polytetrafluoroethylene (ePTFE). All patients received antiinflammatory drugs in the perioperative period. Fever, white blood cells and platelet count, serum concentrations of cytokines (interleukin 6 [IL-6], tumor necrosis factor alpha [TNF-alpha], interleukin 8 [IL-8], acute-phase proteins high-sensitivity C-reactive protein [hsCRP] and alpha1-antitrypsin [alpha1-antitrypsin]), and complement protein (C3a) were measured preoperatively and 1, 3, 6, 24, 48, and 72 hours after aneurysm exclusion. One patient in each group had a systemic inflammatory response syndrome with 2 of the systemic inflammatory response syndrome (SIRS) criteria. No other complication associated with inflammation were present in any patient. Fever was more frequent in group A patients. Increases of white blood cells and serum concentrations of IL-6, TNF-alpha, hsCRP, alpha1-antitrypsin, and C3a and decrease of platelet count were recorded in both groups, but no statistically significant difference between them was recorded. However, serum concentrations of IL-8 were significantly higher in group A patients 24 hours postoperatively (p = 0.01). No significant difference was apparent in the biological response between patients receiving a polyester or an ePTFE stent graft, except for fever and serum concentrations of IL-8.

Acute-Phase Proteins↗

Enhancing effect of antitumor polysaccharide from Astragalus or Radix hedysarum on C3 cleavage production of macrophages in mice.

Effects of Astragalus polysaccharide (APS) and Radix hedysari polysaccharide (RHPS) on the third component of complement (C3) cleavage production of macrophages in ICR mouse were investigated by the immunofluorescent method. By two hours after intraperitoneal injection of 500 mg/kg of APS or RHPS, there was an obvious increase in the deposition of C3 on peritoneal macrophages. When APS or RHPS was injected 5 times (1 time/day), the proportion of C3 positive macrophages was more than that of 1-time injection. At the same time, the number of peritoneal macrophages also increased. Since C3 cleavage product occurred with the activation of C3, the results suggest that the immuno-potentiating action of APS and RHPS may be related to the activity of mouse complement C3.

Animals↗