Low complement levels and opsonic activity in hepatic hydrothorax: its relationship with spontaneous bacterial empyema.

GOALS: To analyze the pleural fluid factors that might cause spontaneous bacterial empyema (SBEM) in patients with cirrhotic hydrothorax. BACKGROUND: Pathogenic mechanism of SBEM of cirrhotic patients is probably similar to that of spontaneous bacterial peritonitis, but local factors affecting pleural fluid have not been studied. STUDY: Determination of C3, C4, and opsonic activity levels of pleural fluid in a cohort of patients with pleural effusions of different causes. RESULTS: Forty-eight patients had hepatic hydrothorax; 8, heart failure and 45, exudates (9, tuberculosis; 21, malignancies; 10, other). Of the 48 cirrhotic patients, 15 developed SBEM on admission. The pleural fluid of cirrhotic patients showed significantly lower levels of total protein, complement, and opsonic activity than did the fluids of patients with other causes of SBEM. Patients who developed SBEM had lower concentrations of pleural fluid total protein and C3 and had a higher Child-Pugh score than patients who did not develop the infection. CONCLUSION: Cirrhotic patients with hepatic hydrothorax have lower pleural fluid opsonic activity and C3 levels than those found in the pleural fluid of patients with other causes. Patients who develop SBEM have lower levels of pleural fluid C3, pleural fluid total protein, and a higher Child-Pugh score than those who do not develop SBEM.

Efecto in vitro de la tuftsina (L-prolil-L-arginina) en la capacidad oxidativa de los polimorfonucleares en niños recién nacidos pequeños para su edad gestacional / Tuftsin (L-prolil-L-arginina) has the function of activating cellular macrophages and polymorphonucleras in newborns

Se comunican los resultados de un trabajo realizado para determinar el efecto de la tuftsina sintética en la capacidad oxidativa de células polimorfonucleares de niños recién nacidos pequeños para su edad gestacional, ya que se ha demostrado que está disminuida la actividad fagocítica de macrófagos y polimorfonucleares (PMNs) comparada con las células de niños con peso adecuado para su edad gestacional. En los resultados se observó un efecto de estimulación de la capacidad oxidativa de los PMNs con tuftsina sintética a través de incrementar la reducción de nitroazul de tetrazolio, lo que sugiere que la baja capacidad oxidativa de las células de recién nacidos pequeños para su edad gestacional no se relaciona con un defecto intrínseco celular

Chemoattractant and opsonic activity in ascitic fluid. A study in 47 patients with cirrhosis or malignant peritonitis.

We studied prospectively the ascitic fluid of 47 patients. Thirty-five were cirrhotics (group A) and 12 had malignant peritonitis (group B). All ascitic fluid samples were initially uninfected. We measured opsonic activity by a chemiluminescent assay, and chemoattractant activity by the under agarose technique. We also measured ascitic concentrations of C3, C4, fibronectin, C-reactive protein, immunoglobulins G, A and M and total proteins. All patients were followed throughout the presence of ascites. None of the group B patients developed peritoneal infection, nor did 23 of the group A patients (group A2). Twelve group A patients (group A1) developed spontaneous bacterial peritonitis (SBP), four of them with recurrence. All indices except immunoglobulins A and M were significantly different between group A and group B patients. Comparing group A1 and group A2, only chemoattractant activity and concentrations of total proteins and C3 were significantly lower in group A1. Using a multivariate analysis with Cox's model, only C3 concentration had an independent predictive value for occurrence of SBP in cirrhotic patients.

Pseudomonas hyperimmune globulin passive immunotherapy for pulmonary exacerbations in cystic fibrosis.

We studied the effect of an intravenously administered gamma globulin [Ps-ivIG] enriched fivefold over conventional ivIG for Pseudomonas aeruginosa lipopolysaccharide [PA LPS] antibodies on ten patients with cystic fibrosis [CF] aged 19-32 years during hospitalization for pulmonary deterioration. All were colonized with greater than or equal to 1 PA phenotype resistant to all antibiotics at the time of admission and they received 500 mg/kg Ps-ivIG intravenously as a single dose in addition to conventional treatment, including antibiotics and chest physiotherapy. No adverse effects occurred. Circulating immune complexes and complement levels remained unchanged from baseline. Serum levels of anti-PA LPS IgG, as measured by ELISA for eight PA LPS immunotypes, increased to 244 +/- 65% (mean +/- SE) of baseline levels 1 hour post-infusion (P less than 0.01), remained significantly elevated during a mean hospital stay of 17 days, and returned to near baseline by follow-up 4 weeks after hospital discharge. Plasma half-life and clearance values were similar to those of other subjects receiving conventional ivIG. Sputum PA density declined from 3.0 to 1.2 x 10(8) cfu/mL 1 week post-infusion (P approximately equal to 0.05), and returned to baseline at follow-up. Serum anti-PA opsonic activity increased after infusion (P less than 0.01), but returned to baseline by 72 hours. Clinical scores improved from admission to discharge (P less than 0.005) without decline at follow-up. Forced vital capacity [FVC] and forced expiratory volume in one second [FEV1] increased from admission to discharge (P less than 0.01 and P less than 0.05, respectively) without decline at follow-up. Using autologous historical control data, standard hospital therapy without Ps-ivIG resulted in no improvement in FVC or FEV1, and a subsequent decline in these parameters (P less than 0.05 for each) during a similar follow-up period. This occurred despite the fact that half the patients did not have antibiotic-resistant PA on the control admission. We conclude that Ps-ivIG is a safe adjunctive therapy for pulmonary exacerbations in moderately ill cystic fibrosis patients colonized with resistant PA, and may be associated with both greater and more prolonged improvement in pulmonary function than standard therapy alone.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) and opsonization synergistically enhance leukotriene B4 (LTB4) synthesis induced by phagocytosis in human neutrophils.

Normal human blood neutrophils were studied for their capacity to synthesize leukotriene B4 (LTB4) and its omega-oxidized metabolites after phagocytosis of zymosan. Phagocytosis of serum-opsonized particles led to a higher release of LTs than did unopsonized zymosan. The most striking effect of phagocytosis was observed when neutrophils were primed with granulocyte-macrophage colony-stimulating factor (GM-CSF): opsonization and GM-CSF synergistically increased LTB4 synthesis by neutrophils.

[Plasma fibronectin in surgical patients].

Plasma fibronectin (PFN) was measured to clarify the role in patients who underwent major surgery. There was no statistical difference between male and female, benign and malignant disease, or more than 60 years old and less. Compared to the preoperative values of PFN, the postoperative ones showed significant decrease immediately after operation, this decrease was attributed to massive bleeding and prolonged surgical procedure. In contrast to the previous papers reporting as an opsonin, no such results were obtained. However, there was close relationship between PFN and antithrombin III, alpha 2-plasmin inhibitor, factor XIII, retinol binding protein and transferrin, which represent hemostasis, fibrinolytic system and nutritional status, respectively. To determine whether PFN acts as an opsonin, heparin (opsonin cofactor) was given to Wistar rats in various conditions. There was no significant difference between PFN in those rats and in controls. These results indicate that postoperative PFN is a good parameter of glycoprotein in hemostasis, fibrinolytic system, and rapid turnover protein, not of an opsonin.

[Opsonic cooperation in the neutrophil phagocytosis system in acute myeloblastic leukemia]. / Opsonicheskaia kooperatsiia v sisteme neitrofil'nogo fagotsitoza pri ostrom mieloblastnom leikoze.

Investigation of opsonic activity of the serum (OAS) and indices of neutrophil phagocytosis in 35 patients with acute myeloblastic leucosis (AML) and 20 healthy subjects indicates that reduction of OAS in AML results in disturbances of the processes of reception and absorption of microorganisms by granulocytes while insufficient activation of oxygen-dependent metabolism favours disorders of bactericidal activity of neutrophils.

The three different phases of reticuloendothelial system phagocytic function in rats with liver injury.

In the present study, reticuloendothelial system (RES) phagocytic function of rats with partial hepatectomy or experimentally induced liver cirrhosis was investigated by determining the phagocytic index, the opsonic index, and uptake rate in liver, spleen, and lung of a 51Cr-labeled endotoxin-injected rat. In both the partially hepatectomized and the cirrhotic rats, all three indicators varied markedly according to the elapsed period since liver injury. The changes in RES phagocytic function were classified into three different phases: compromised, compensatory, and enhanced. The compromised phase, consisting of a decrease in the phagocytic index, was observed during the first 24 hr after 67% hepatectomy and in advanced liver cirrhosis. This represented the failure of RES phagocytic function. The compensatory phase, in which the phagocytic index was maintained at nearly normal levels mainly by a compensatory enhancement in the opsonic index, was seen during the first to second postoperative day and in moderate liver cirrhosis. The enhanced phase, with a high phagocytic index, was observed from Day 4 to approximately Day 14 after surgery, and in the cases of mild liver damage. In the compromised and compensatory phases, the liver uptake rate was significantly decreased compared with the control. However, the uptake in the spleen and lung were markedly increased. In conclusion, the phagocytic function of the RES was significantly affected to a degree which changed with the extent of liver damage.

Response of CAPD patients with a high incidence of peritonitis to intraperitoneal immunoglobulin therapy.

This study has demonstrated that in the majority of high-peritonitis-incidence (PI) CAPD patients the defective opsonic activity levels in the peritoneal dialysis effluent (PDE) are restored for 3 weeks by a 12 g IP injection of immunoglobulins (Ig). Further studies showed that in a minority of high-PI CAPD patients who also had low PDE IgG and opsonic activity levels, IP Ig therapy did not significantly reduce the PI. To evaluate this phenomenon we utilized this therapy in 20 high-PI CAPD patients undergoing IP Ig therapy for an average of 24 months daily for 3 weeks (12 g every 3 weeks) and analyzed: 1) PDE IgG levels; 2) PDE opsonic activity; 3) peritoneal macrophage (PM0) membrane-bound IgG; 4) PDE Interleukin-1 (IL-1) levels; 5) PM0 membrane Fc receptor number. The results showed that in the 15 patients in whom Ig therapy reduced PI, there were long-lasting increases in the PDE IgG, opsonic activity and IL-1 levels, as well as a normal PM0 Fc receptor number and PM0 reversibly bound infused Ig. Conversely, the five patients in whom the IP Ig did not reduce the PI showed only transient increases in PDE IgG and opsonic activity levels, no PDE IL-1 increase, and the PM0 were deficient in Fc receptors and, therefore, unable to take up the infused Ig. We conclude that in high-PI CAPD patients there are different peritoneal immune defense abnormalities and that their identification is, therefore, important for the correct choice of therapy to improve these defects.

[Postoperative debasement of the host-defense system and wound healing in elderly patients, and nutritional support as a therapeutic implication].

Plasma opsonic activity and several kinds of plasma protein levels in each pre- and post-operative days were compared between older 25 cases (70 years or older) and 31 younger cases (59 years or younger) which both underwent elective abdominal surgery. The activity to opsonize E. coli 075 as a host-defense capacity was reduced on postoperative days 1,3 and returned gradually in either young or elderly. But opsonic activity levels in elderly patients in each postoperative days were significantly lower than in younger group. Plasma albumin level and rapid turnover protein levels in elderly patients tended to be lower than those in younger group through the postoperative period but statistically not significant. Plasma levels of C3 and fibronectin overshot the each preoperative value on days 5-7, after sharp reduction on day 1 in younger patients, but those in elderly didn't. Opsonic activity levels were closely correlated with plasma protein levels. Based on these clinical results, the experiments were designed to evaluate the effect of nutritional support in the early postoperative period on opsonic activity and protein synthesis in aged rats. These studies elucidated that early nutritional support brought beneficial effects on opsonic activity and plasma protein levels on postoperative days 7, but not on days 3 in aged rats. This may be because in aged animals, protein synthetic function has not fully recovered in the early postoperative period.

Immunity to malaria in young Gambian children after a two-year period of chemoprophylaxis.

A cohort of 48 Gambian children was protected against malaria by fortnightly administration of Maloprim (pyrimethamine and dapsone) for 2 years between their 3 and 5 birthdays. A matched cohort of 47 children received placebo. During the year following the termination of prophylaxis there was no increase in the frequency of clinical attacks of malaria in the protected children compared with the control children. Antibody levels to circumsporozoite protein were measured by a radioimmunoassay and that to blood-stage antigens by a variety of techniques including an ELISA to whole blood-stage Plasmodium falciparum antigen, immunofluorescent assays (IFAT) to acetone fixed, glutaraldehyde fixed and unfixed parasites, a merozoite inhibition test and an opsonizing assay. Antibody levels were, in general, lower in protected than in control children and several differences between the two groups were statistically significant. When antibody levels were measured by ELISA and IFAT at the end of the following rainy season, when malaria transmission was intense, those in protected children had increased to comparable levels to those found in control children. Our findings suggest that chemoprophylaxis given for 2 years lowers malaria antibody levels but that it does not interfere with the development of protective immunity.

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.

Correlation of serum opsonic activity in cystic fibrosis with colonization and disease state: measurement of opsonins to Pseudomonas aeruginosa by neutrophil superoxide anion generation.

Serum from patients with cystic fibrosis and normal controls was used to opsonize mucoid and nonmucoid Pseudomonas aeruginosa particles. Opsonic activity was then determined by measuring the production of superoxide anion (O2-) from normal neutrophils stimulated with the opsonized particles. Without any opsonization, mucoid P. aeruginosa stimulated significantly more O2- than nonmucoid P. aeruginosa. Responses to nonmucoid P. aeruginosa observed with heat-inactivated serum from patients with cystic fibrosis were significantly higher (p = 0.008) than those observed with heat-inactivated control sera. Comparisons made between patients who were colonized with P. aeruginosa and those who were not showed that heat activated serum from colonized patients had significantly higher levels of opsonic activity than heat inactivated serum from patients who were not colonized. These differences were observed with either mucoid or nonmucoid P. aeruginosa. A negative correlation was also observed between opsonic activity and clinical status measured by Schwachman scores of colonized patients. These data indicate that in patients colonized with P. aeruginosa the deterioration of their clinical status correlated with increased opsonic activity reflected in the oxidative burst response of neutrophils.

[Experimental and clinical studies on the effect of reticuloendothelial system (RES) potentiator in the depressed RES function in cirrhotics].

Among the patients with liver cirrhosis (LC) who undergo the operation, the postoperative complications are not infrequent and sometimes prove fatal. The impaired hepatic function, especially the impaired reticuloendothelial system (RES) function, has been claimed to be a possible pathogenic factor for these complications. The present experimental and clinical studies were undertaken to investigate the RES function and the effect of preoperative OK-432 administration as an RES potentiator in LC. The results are as follows: 1) CCl4-induced LC rats were evaluated for RES global phagocytic function, Kupffer cell phagocytic function, plasma opsonic activity and plasma opsonic substances such as fibronectin, C3 and IgG. All parameters except IgG showed significant depression compared to those values in normal rats. However, the administration of OK-432 (0.1 KE/rat, ip) improved all these depressed parameters. The OK-432 administration also significantly improved the survival following panperitonitis in LC rats. 2) Among 18 LC patients with hepatocellular carcinoma undergoing partial hepatectomy, the RES global phagocytic function, plasma opsonic activity and plasma opsonic substances were evaluated. Same as the experimental study, all parameters except IgG were significantly depressed among the LC patients compared to those values in the patients with normal liver. However, the preoperative OK-432 administration (5 KE/day sc for 4 days) significantly improved these parameters and consequently decreased the postoperative complications. These results indicate that the preoperative RES activation by the OK-432 was effective and useful for the prevention of the postoperative complications in the LC patients.

[Chemiluminescence of whole blood--analysis of the kinetics and application of clinical examination of phagocytic functions of whole blood from various type of diseases].

Luminol-dependent chemiluminescence of whole blood (whole blood CL) was developed to estimate the phagocytic function of granulocytes and the serum opsonin activity simultaneously. Whole blood (0.1 ml) was examined directly and results were obtained within 20 minutes. Phagocytic function of granulocytes can be estimated from the peak CL of whole blood and the number of granulocytes in a specimen, and the opsonin activity from the amount of time the peak CL is shown after the addition of nonopsonized CL inducer (nonopsonized zymosan). Subsequently, whole blood CL was measured to evaluate phagocytic functions in children with disease. Patients with chronic granulomatous disease showed no CL, and one of their mothers (1/3) showed low CL, suggesting she is a carrier. Two patients with hypocomplementemia (SLE and chronic nephritis) showed low serum opsonin activity, and phagocytic function of their granulocytes was enhanced. In cord blood and newborn infants serum opsonin activity was low, and phagocytic function of granulocytes was slightly decreased in cord blood but not in newborn infants. Patients with systemic bacterial infections showed an increased phagocytic function of granulocytes. Anti-cancer drugs decreased serum opsonin activity in children with leukemia or lymphoma. The children treated with L-asparaginase had very low opsonin activity, suggesting the drug inhibits complement synthesis. The measurement of whole blood CL was useful for monitering the phagocytic functions of blood after granulocyte transfusion.

Postnatal maturation of pulmonary antimicrobial defense mechanisms in conventional and germ-free lambs.

The phagocytic and bactericidal capacities of ovine bronchoalveolar lavage (BAL) cells were investigated as a function of postnatal age. In addition, age-related changes in the elaboration by alveolar macrophages of chemotaxins for neutrophils, concentrations of BAL fluid and serum immunoglobulins, and serum opsonic capacity were determined. BAL cells exhibited major changes in morphology, composition, and in vitro proliferation during the 1st postnatal wk. Studies in germ-free lambs indicated that the antigenic burden of the ambient environment markedly influenced the concentration of BAL neutrophils but had no effect on the influx, phagocytic, and proliferative activities of alveolar macrophages. Phagocytic and bactericidal functions of BAL cells improved rapidly during the 1st postnatal wk, then declined, and did not reattain adult levels until day 180. The capacity of alveolar macrophages to elaborate chemotaxins for neutrophils was deficient at day 8, but not at subsequent ages. The concentration of BAL IgG1 increased until day 8, fell at day 21, and then continued to increase gradually. IgA was not detected in BAL until day 21 and increased rapidly thereafter. Serum opsonic capacity at days 1 and 4 was comparable to that of adult serum, but sera from days 8 to 42 showed a marked reduction in opsonic capacity. Pulmonary antimicrobial defenses in neonatal sheep were thus found to be deficient to some degree throughout the first 3 months of life. It was not until day 180 that the parameters investigated in this study approximated those of adult sheep.

Intracellular survival of Candida albicans in peritoneal macrophages from chronic peritoneal dialysis patients.

Candidal peritonitis is a tenacious infection in patients undergoing chronic peritoneal dialysis. Since little is known about host defenses of the human peritoneal cavity against fungi, we investigated the interaction of peritoneal macrophages (PM phi) from uninfected dialysis patients with Candida albicans blastospores. Chemiluminescence (CL) techniques were used to assess the respiratory burst activity of these cells, and candidacidal activity was evaluated with a fluorochrome microassay. In sharp contrast to peripheral blood polymorphonuclear leukocytes (PMNs) from healthy donors, which gave a brisk luminol-enhanced CL response to opsonized blastospores and killed 35% of cell-associated organisms, PM phi produced barely detectable luminol-enhanced CL and killed only 13% of intracellular Candida. These findings appeared to be associated with a decreased level of myeloperoxidase in PM phi. The mechanism of intracellular survival of C albicans also appeared to be related to relatively poor triggering of superoxide production during phagocytosis of viable blastospores. The CL response of PMNs to C albicans was opsonin-dependent, and peritoneal dialysis effluent was devoid of opsonic activity. These studies suggest that local cellular and humoral mechanisms of defense are inadequate for protection of peritoneal dialysis patients against candidal peritonitis.

Técnicas para evaluar la capacidad fagocítica, opsonica y bactericida del leucocito polimorfonuclear / Techniques to evaluate the phagocytic, opsonizing and bactericidal ability f the polymorphonuclear leukocyte

Para estudiar la función in vitro del leucocito PMN se estandarizaron las técnicas inmunológicas de capacidad fagocítica, opsónica y bactericida. La capacidad fagocítica se midió en sangre total, incubando leucocitos del sujeto en estudios en presencia de su propio plasma con estafilococos aureus muertos y la actividad opsónica se determinó incubando células de un individuo sano con plasma del sujeto en estudio. Los frotis teñidos se leyeron al microscopio de luz, y los resultados se expresaron como índice fagocítico, esto es número de bacterias ingeridas por 100 PMN. La capacidad bactericida se estudió, incubando una suspensión de leucocitos con bacterias (E. coli) en rotación continua a 37-C, basada en la técnica de Quie y cols. Se tomaron alícuotas para recuento de colonias por dilución en placa a los tiempos 0,20, 60 y 120 minutos. Los resultados se expresaron como índice bactericida, esto es, número de bacterias vivas a los 120 minutos/número de bacterias vivas a los 20 min. Para optimizar las técnicas se analizaron los siguientes parámetros: número de leucocitos/ml de sangre o suspensión, concentración y cepa de referencia de E. coli/ml de suspensión, período de incubación, reproducibilidad de la técnica, variabilidad intraindividual e interindividual y en el tiempo. Una vez estandarizados dichos parámetros, los índices de capacidad fagocítica y opsónica fueron de 9.2 ñ 0.85 y 8.1 ñ 0.6 (x ñ ESM), respectivamente. El índice promedio de la actividad bactericida fue de 0.14 ñ 0.05. Las pruebas fueron discriminatorias en pacientes con sospecha clínica de alteración de la capacidad funcional del leucocito

In-vivo effects of clindamycin on neutrophil function.

Neutrophil functions were evaluated in 13 normal subjects who had received 300 mg of clindamycin orally four times each day for two days. The mean serum concentration of clindamycin was 1.6 mg/l. Intracellular killing of a clindamycin-resistant strain of Staphylococcus aureus increased from 38% to 45%, P less than 0.005, during clindamycin therapy. In contrast, clindamycin therapy did not significantly alter chemotaxis, phagocytosis, chemiluminescence of neutrophils, or the ability of serum to generate chemotactic factor and opsonize particles of yeast. The potentially synergistic relationship between clindamycin and neutrophils may prove to be valuable for the treatment of staphylococcal infections in patients with defects in oxygen-dependent mechanism of neutrophil-mediated bacterial killing such as in chronic granulomatous disease.

Impairment of opsonic function in children with hematologic malignancy during remission induction therapy.

The activity of complement-mediated opsonin was measured by the whole blood chemiluminescence method in 17 children with hematologic malignancy (including 6 with ALL, 7 with ANLL and 4 with non-Hodgkin's lymphoma) during remission induction therapy. The activity of opsonin, which was at the normal level before chemotherapy, decreased in all of the children during the therapy. This phenomenon was especially marked in the children treated with L-asparaginase. Although no clear relationship was found between the decrease in opsonin activity and the susceptibility to infection, it was confirmed that in 4 children having an episode of sepsis or septic fever, the infection started when the granulocyte decreased to the nadir, and simultaneously the activity of opsonin decreased. Therefore, it may be reasonable to suspect the decrease in opsonin activity when treating children with such infections.