C5a stimulates secretion of tumor necrosis factor from human mononuclear cells in vitro. Comparison with secretion of interleukin 1 beta and interleukin 1 alpha.

We have demonstrated that purified C5a is a potent stimulus to human PBMC secretion of TNF-alpha, IL-1 beta, and IL-1 alpha, which proceeds in a dose-dependent fashion. At a given concentration of C5a, TNF-alpha and IL-1 beta secretion did not differ significantly; both were secreted in significantly greater quantity than IL-1 alpha. Clinical conditions such as Gram-positive and Gram-negative bacterial infections, trauma, and immune complex diseases activate complement. Through the mediation of TNF and IL-1 secreted in response to C5a, these diverse disorders can share common features of fever, coagulopathy, acute phase protein production, and disordered metabolism.

Interleukin 1 induces a shock-like state in rabbits. Synergism with tumor necrosis factor and the effect of cyclooxygenase inhibition.

In addition to activating T and B lymphocytes, interleukin 1 (IL-1) induces several hematologic and metabolic changes typical of host responses to infection and injury. We now report a new biological property, namely, the induction of hypotension. Rabbits given a single intravenous injection of recombinant human IL-1-beta (5 micrograms/kg) rapidly developed decreased systemic arterial pressure, which reached the lowest levels after 50-60 min and slowly returned to pre-IL-1 values after 3 h. Associated with the hypotension, systemic vascular resistance and central venous pressure fell, while cardiac output and heart rate increased. These responses were prevented by ibuprofen given 15 min before the IL-1. A bolus injection of IL-1 followed by a 2-h infusion sustained the hypotension and was associated with leukopenia and thrombocytopenia. Ibuprofen given at the mid-point of the infusion reversed the changes in all hemodynamic parameters, but had no effect on the leukopenia or thrombocytopenia. Tumor necrosis factor (TNF) also induced a shock-like state in rabbits. When the dose of IL-1 or TNF was reduced to 1 microgram/kg, no hemodynamic changes were observed; however, the combination of these low doses of both cytokines resulted in a profound shock-like state including histological evidence of severe pulmonary edema and hemorrhage. Pretreatment with ibuprofen prevented the hemodynamic, leukocyte, and platelet changes induced by the low-dose cytokine combination, and ameliorated the pulmonary tissue damage. These results demonstrate that IL-1, like TNF, possesses the ability to induce hemodynamic and hematological changes typical of septic shock, and that the combination of IL-1 and TNF is more potent than either agent alone. These effects seem to require cyclooxygenase products, and suggest that intravenous cyclooxygenase inhibitors may be of therapeutic value in patients with IL-1/TNF-mediated shock.

Concentrations of immunoreactive human tumor necrosis factor alpha produced by human mononuclear cells in vitro.

The concentrations of tumor necrosis factor (TNF) produced by human peripheral blood mononuclear cells (MNC) were measured using a radioimmunoassay (RIA) for human TNF. This was developed using a rabbit antiserum against human recombinant TNF (Hu rTNF), and Hu rTNF labeled with Na125I by a modification of the chloramine T method. This RIA does not detect human lymphotoxin, interleukin-1 alpha or beta, interleukin 2, interleukin 6, interferon alpha or gamma, granulocyte-macrophage-colony stimulating factor, and C5a des arg. A good correlation (r = 0.89) was found between the RIA and the cytolytic bioassay for TNF. The sensitivity of the RIA is between 3 and 78 pg/ml (median 11 pg/ml). The mean concentration of TNF in 24-h culture supernatants of human MNC exposed to different concentrations of lipopolysaccharide (LPS) was found to increase in dose-dependent fashion and then level off between 50 and 100 ng/ml. The concentrations of IL-1 beta and alpha detected by specific RIAs in these supernatants were between 0.2 and 19 ng/ml and 0.04 and 1 ng/ml, respectively. The amount of TNF produced by human MNC in vitro was determined in a cohort of 50 normal volunteers. Without exogenous stimuli, TNF concentrations were almost always below the detection limit; with 0.5 ng/ml LPS, the median concentration of TNF was 2 ng/ml, and with PHA the median was 3.8 ng/ml. In cultures performed in the presence of indomethacin significantly (p less than 0.005) more TNF was produced. Using this RIA, we could detect TNF in the circulation of mice injected with Hu rTNF. When plasma samples of patients with febrile illnesses were added directly to the RIA, TNF was not detectable, with the exception of patients with malaria. These studies demonstrate the range and sensitivity of LPS-induced and mitogen-induced production of immunoreactive TNF by human MNC in vitro without interference of similar cytokines in bioassays.

[Hemodynamic changes in acute pancreatitis].

Hemodynamics studies were carried out from days 1-5 following the onset of illness in 7 patients with severe pancreatitis (group A) and in 7 patients with moderate pancreatitis (group B). Patients in both groups had a higher cardiac index (CI) and a lower systemic vascular resistance (SVR) than normal patients during 5 days of illness, and patients in group A had a higher CI of 5.38 +/- 0.841 x min/m2 and lower SVR of 889 +/- 234dyn.sec/cm5 than those in group B on day 4. Patients in group A had a lower pulmonary vascular resistance than normal patients on days 1, 3, and 4, but those in group B did not show this hemodynamic change. Group A patients had a higher pulmonary wedge pressure of 11.9 +/- 8.4mmHg and depressed left ventricular stroke work index of 59.8 +/- 17.8g.m/m2 as compared with group B (5.6 +/- 3.4mmHg, 77.7 +/- 23.6g./m2, respectively). These findings indicate that a hyperdynamic hemodynamic state may exist in the early stages of moderate and severe pancreatitis and myocardial depression may be evident in severe pancreatitis.

[Evaluation of splenectomy and possible metastasis from gastric cancer, with reference to nonspecific immunological parameters and postoperative survival].

For the complete removal of metastasized lymph nodes at splenic hilus in the operation for gastric cancer, the splenectomy has been widely accepted. In order to reveal justifiable of such splenectomy, nonspecific immunological parameters and postoperative survival were compared between the groups of splenectomized (S) and nonsplenectomized cases (N) in the same stage of gastric cancer. The immunological parameters in N were more stable after surgery than those in S. As to the postoperative survival there was no significant difference. The survival rates showed no definite difference. The postoperative platelet count of S was higher than that of N. It may be concluded that the splenectomy should be avoided unless the metastases are remarkable.

Postoperative metabolic alkalosis following general surgery: its incidence and possible etiology.

A prospective clinical study was performed on 293 patients, in order to elucidate the abnormalities in acid-base balance following general surgery. Six arterial blood gas and pH determinations were taken from each patient before surgery and on postoperative days zero, one, three, five and seven. A total of 1699 determinations were obtained. Although the majority of patients (87.5 per cent) had a normal acid-base balance before surgery, a postoperative metabolic alkalosis was seen in 50.5 per cent of the patients. However, there was an extremely low incidence of other postoperative acid-base abnormalities, apart from a transient increase in metabolic acidosis on the operative day. A significantly high mortality rate (32.3 per cent) was observed in 31 patients who had continuous metabolic alkalosis during the postoperative period. An excessive bicarbonate load resulting from the administration of fresh frozen plasma following surgery was strongly suggested as one of the major causes of postoperative metabolic alkalosis. Further investigation is required to elucidate the mechanism of the generation of metabolic alkalosis induced by the postoperative bicarbonate load in surgical patients.

Interleukin-1 induces a shock-like state in rabbits: synergism with tumor necrosis factor and the effect of cyclooxygenase inhibition.

In addition to activating T and B lymphocytes, interleukin-1 (IL-1) induces several hematologic and metabolic changes typical of host responses to infection and injury. We now report a new biological property, namely, the induction of hypotension. Rabbits given a single intravenous injection of recombinant human IL-1-beta (5 micrograms/kg) rapidly developed decreased systemic arterial pressure with the lowest levels after 50-60 min. Associated with the hypotension, systemic vascular resistance and central venous pressure fell while cardiac output and heart rate increased. These responses were prevented by intravenous ibuprofen given 15 minutes prior to the IL-1. A bolus injection of IL-1 plus a 2 hour infusion sustained the hypotension and was associated with leukopenia and thrombocytopenia. Ibuprofen given at the mid-point of the infusion reversed the changes in all hemodynamic parameters. Tumor necrosis factor (TNF) induced a more profound shock-like state in rabbits. When the dose of IL-1 or TNF was reduced to 1 microgram/kg, no hemodynamic changes were observed; however, the combination of these low doses of both cytokines resulted in a profound shock-like state. Ibuprofen prevented the hemodynamic, leukocyte and platelet changes induced by the low-dose cytokine combination. These results demonstrate that IL-1, like TNF, possesses the ability to induce hemodynamic and hematological changes typical of septic shock, that the combination of IL-1 and TNF is more potent than either agent alone, and that cyclooxygenase products are involved in IL-1/TNF-mediated shock.

Induction by toxic-shock-syndrome toxin-1 of a circulating tumor necrosis factor-like substance in rabbits and of immunoreactive tumor necrosis factor and interleukin-1 from human mononuclear cells.

A shock-like syndrome was induced in rabbits by administering toxic-shock-syndrome toxin-1 (TSST-1); tumor necrosis factor (TNF)-like activity was detected in sera of rabbits 3.5 h after injection, as measured by cytotoxic effects on the tumorigenic L929 murine fibroblast cell line. Appearance of this activity in sera coincided with onset of significant shock-related hemodynamic changes. TSST-1 stimulated release of TNF-like material from rabbit mononuclear cells in culture. Human mononuclear cells also secreted a cytotoxic substance shown to be TNF by radioimmunoassay. Maximal TNF secretion was higher in human mononuclear cells stimulated with TSST-1 than in those stimulated with bacterial lipopolysaccharide. Lipopolysaccharide, however, was a more potent inducer of interleukin-1 alpha and interleukin-1 beta from the same cells than was TSST-1. Because TNF and interleukin-1 act synergistically during induction of a shock-like state, these results suggest that part of the TSST-1-induced shock is due to production of interleukin-1 and TNF.

Interleukin-1 induces tumor necrosis factor (TNF) in human peripheral blood mononuclear cells in vitro and a circulating TNF-like activity in rabbits.

Recombinant human interleukin-1 (IL-1), injected into rabbits, induces the synthesis of endogenous IL-1. Also, IL-1 induces its own gene expression and synthesis in human peripheral blood mononuclear cells (PBMC). In this study, tumor necrosis factor-alpha (TNF-alpha) production by PBMC of 40 individuals stimulated with IL-1 alpha or IL-1 beta was determined by specific radioimmunoassay (RIA). After 3 h of PBMC incubation with IL-1, TNF-alpha mRNA was detected. IL-1 alpha stimulated both IL-1 beta and TNF-alpha, but there was no correlation in the amount of TNF-alpha or IL-1 beta synthesized in the PBMC of 29 individuals. IL-1-stimulated adherent cells produced approximately 50% more TNF-alpha than did unfractionated PBMC. Coincubation with interferon-gamma (IFN-gamma) did not change the amount of IL-1-induced TNF-alpha, whereas in the same culture IFN-gamma inhibited (greater than 70%) IL-1-induced IL-1 production. Endogenous pyrogen and TNF-like activity were detected in the sera of rabbits 3.5 h after injection of either IL-1 alpha or -1 beta. These studies demonstrate that IL-1 induced TNF-alpha production in vivo and in vitro.

Role of interleukin 1 and tumor necrosis factor on energy metabolism in rabbits.

A study of the combined effects of intravenous infusion of the recombinant cytokines beta-interleukin 1 (IL-1) and alpha-tumor necrosis factor (TNF) on energy substrate metabolism in awake, conditioned, adult rabbits was performed. After a 2-h basal or control period, 48-h fasted rabbits were administered TNF and IL-1 as a bolus (5 micrograms/kg) followed by a continuous intravenous infusion (25 ng.kg-1.min-1) for 3 h. Significant increases in plasma lactate (P less than 0.01), glucose (P less than 0.01), and triglycerides (P less than 0.05) occurred during the combined infusion of IL-1 and TNF, whereas neither cytokine alone had no effect. There was a 33% increase in the rate of glucose appearance (P less than 0.05), but glucose clearance was not altered compared with the control period. Glucose oxidation increased during the combined cytokine infusion period and glucose recycling increased by 600% (P less than 0.002). Lactic acidosis and decreased oxygen consumption, as a result of the cytokine infusions, indicated development of anaerobic glycolytic metabolism. A reduction in the activity state of hepatic mitochondrial pyruvate dehydrogenase (65 vs. 82% in control animals, P less than 0.05) was consistent with the observed increase in anaerobic glycolysis. Thus the combined infusion of IL-1 and TNF in rabbits produces metabolic manifestations seen in severe injury and sepsis in human patients and, as such, may account for the profound alterations of energy metabolism seen in these conditions.

C5a induction of human interleukin 1. Synergistic effect with endotoxin or interferon-gamma.

Interleukin-1 (IL-1) is a potent cytokine which possesses the ability to mediate systemic acute phase responses as well as local tissue inflammation. In these studies, we have examined the ability of C5a and C5a des Arg to induce IL-1 production in vitro. Human C5a and C5a des Arg were purified to homogeneity and were found to stimulate IL-1 release from freshly obtained human mononuclear cells into the extracellular medium. Only 2 hr of exposure to the purified complement components were necessary in order to stimulate IL-1 production. The minimal concentration of C5a required was 25 ng/ml, whereas 125 ng/ml of C5a des Arg induced comparable amounts of IL-1. This dose relationship was maintained at higher concentrations (150 ng/ml vs 750 ng/ml, respectively). That the effect was due to the anaphylatoxins themselves, and not endotoxin contamination, was shown by negative Limulus amebocyte lysate tests and employing preincubation of C5a/C5a des Arg with polymyxin B. The latter blocked a wide dose range of endotoxin-stimulated IL-1 production. However, when endotoxin was added to C5a or C5a des Arg, significant synergism in the stimulation of IL-1 production was observed, occurring at various concentrations of either agent. A similar synergism with C5a/C5a des Arg was seen with interferon-gamma. In these studies, IL-1 production was measured by bioassay employing cloned D . 10 . G4 . 1 murine T cells and by radioimmunoassay for human IL-1 beta; using C5a/C5a des Arg as stimulants, there was a high degree of correlation (r = 0.82) between the two assays. Since traumatic, infectious, and inflammatory diseases may result in the simultaneous appearance of these stimuli, the synergism described herein is likely to be clinically relevant.

Usefulness and limitations of ultrasonography in the initial evaluation of blunt abdominal trauma.

BACKGROUND: In the assessment of blunt abdominal trauma, the reliability of ultrasonography (US) in identifying individual organ injuries remains uncertain, in spite of its usefulness in detecting hemoperitoneum. This study was designed to evaluate the overall diagnostic value of US, including identification of individual organ injuries. METHODS: The accuracy of US in the detection of intra-abdominal injuries and the identification of individual organ injuries was evaluated in 1,239 patients seen during a 15-year period. Accuracy was based on detection of intraperitoneal fluid, free air, or irregular parenchymal lesions. RESULTS: For the detection of injuries, US was 94.6% sensitive, 95.1% specific, and 94.9% accurate. Individual organ injuries were identified with sensitivities of 92.4, 90.0, 92.2, 71.4, and 34.7% for the liver, spleen, kidneys, pancreas, and intestine, respectively. CONCLUSION: US is reliable for the detection of injuries and the identification of solid-organ injuries despite its poor sensitivity for intestinal injuries.