Evidence for the involvement of interleukin 10 in the differential deactivation of murine peritoneal macrophages by prostaglandin E2.

Among other effects, prostaglandins (PG) of the E series are known to inhibit several acute and chronic inflammatory conditions in vivo and proinflammatory cytokine production by activated macrophages in culture. The research presented here demonstrates that the inhibitory effect of PGE2 on tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) production by lipopolysaccharide (LPS)-stimulated murine peritoneal macrophages involves IL-10. In a dose-dependent manner, PGE2 inhibits LPS-induced release of TNF-alpha and IL-6, but not of lactate or nitric oxide. The decrease in the level of these cytokines is inversely proportional to the increase in immunoreactive IL-10. This differential inhibitory effect of PGE2 is mimicked by agents that elevate intracellular levels of cAMP, but not cGMP. Neutralizing anti IL-10 antibody but not neutralizing antibodies against other macrophage secretory products (IL-6, leukemia inhibitory factor, and transforming growth factor beta [TGF-beta]), significantly reverse the potent inhibitory effect of PGE2. In vivo, the administration of PGE2 before LPS challenge significantly reduces circulating TNF-alpha and IL-6 levels. Anti-IL-10 antibody substantially enhanced the LPS-induced TNF-alpha and IL-6 levels in mice that received either LPS alone or LPS plus PGE2. These results suggest that the anti-inflammatory effect of PGE2 on mononuclear phagocytes is mediated in part by an autocrine feedback mechanism involving IL-10.

cAMP-elevating agents suppress dendritic cell function.

The administration of cAMP-elevating agents affects a number of autoimmune and inflammatory conditions. Because dendritic cells (DCs) play a pivotal role in autoimmunity and inflammation, the isolated effects of cAMP-elevating agents on the function of DCs was examined. In a dose-dependent manner, 8-Bromo cAMP, prostaglandin E(2), and 3-isobutyl-1-methylxanthine inhibited tumor necrosis factor alpha release and suppressed antigen presentation by DCs. The same effect was observed with rolipram, a specific inhibitor of phosphodiesterase type 4, but not with inhibitors of other phosphodiesterases. The decreased antigen presentation by DCs was associated with an enhanced production of interleukin (IL)-10 and with lower major histocompatibility complex type II (MHC II) expression. Furthermore, the inhibition of antigen presentation and MHC II expression was significantly reversed by treatment of DCs with neutralizing antibody against IL-10, suggesting the involvement of an IL-10-dependent mechanism. Taken together, these results might explain why certain cAMP-elevating agents such as rolipram are effective in blocking autoimmunity and inflammation.

[Giant hemangioma in the neck extending into the mediastinum required a second operation after 39 years; report of a case].

An asymptomatic 69-year-old man was admitted to our hospital for an abnormal shadow on the chest X-ray of a medical examination. He had undergone an operation for a hemangioma in the neck (incomplete resection) 39 years before admission. Computed tomography (CT) of the neck and chest revealed an giant cystic mass in the neck extended into the mediastinum, severe deviation of the trachea to the right and fluid collection in the mediastinum. It was considered to be a giant hemangioma with mediastinal hemorrhage by the rupture of the tumor. After the large afferent and efferent vessels were identified by angiography and venography, we performed the resection of the tumor. Pathological examination confirmed the diagnosis of arteriovenous hemangioma.

[Experience with thoracoscopy for rifle gunshot penetrating trauma of the chest; report of a case].

A 57-year-old man came to our hospital by ambulance for a chest injury by a rifle gunshot. He had a penetrating injury of the chest wall, hemopneumothorax and pulmonary laceration. He was managed with chest drainage, oxygen inhalation. His respiratory and cardiac status was stable. However, for the purpose to prevent the development of empyema or pneumonia, and to check the existence of damage of intrathoracic structures by the gunshot injury, thoracoscopy was performed next day. He discharged without postoperative complications 17 days after the injury. Open thoracotomy is reported to be required in only about 10-15% of patients with chest injuries. However, operative indication of the chest injuries may spread in the future with the spread of thoracoscopy and its low invasiveness.

Neutrophils augment the release of TNFalpha from LPS-stimulated macrophages via hydrogen peroxide.

We examined the effect of polymorphonuclear cells on the release of tumor necrosis factor (TNFalpha) in endotoxin-treated macrophages. Human peripheral blood neutrophils were co-cultured with mouse peritoneal macrophages stimulated with lipopolysaccharide (LPS). In a dose-dependent manner, FMLP (n-formyl-methionyl-leucyl-phenylalanine) augmented the release of TNFalpha by LPS-stimulated macrophages in the presence, but not in the absence, of neutrophils. The stimulating effect of neutrophils on macrophages was reversed by catalase, suggesting that the release of hydrogen peroxide from neutrophils was responsible for augmenting macrophage TNFalpha. Moreover, the direct addition of hydrogen peroxide to macrophages resulted in an increased secretion of TNFalpha. In addition, insertion of a porous membrane between the neutrophils and macrophages cancelled the effect, indicating that adherence of neutrophils may be necessary for augmentation of TNFalpha release. In summary, the data suggest that hydrogen peroxide released from stimulated neutrophils may act as an activator of macrophage function by increasing their release of TNFalpha.

The lack of an effect by insulin or insulin-like growth factor-1 in attenuating colon-2-mediated cancer cachexia.

In several studies, the anabolic hormones insulin-like growth factor-1 (IGF-1) and insulin attenuated several metabolic changes associated with cancer cachexia. In the present study, we evaluated the effect of these hormones on the cachexia associated with colon-26 (C-26) tumor. Healthy age-matched and tumor-bearing mice were treated with two daily doses of IGF-1 (50 micrograms/kg in toto), or insulin (1 U in toto). Determinants of cachexia were body and tumor weight, epididymal fat pad and serum glucose concentrations. Neither IGF-1 nor insulin treatment had a significant effect on the cachectic parameters of C-26-bearing mice. These hormones were biologically active, being capable of inducing weight gain in hypophysectomized mice and hypoglycemia, respectively. Although IGF-1 and insulin have been used to treat cancer-related weight loss, the research presented here suggests that the beneficial effect of these hormones is not universal.

Effect of increased systemic venous pressure on thoracic duct and peripheral lymph flow in dogs.

In congestive heart failure, lymph flow from the cannulated thoracic duct is greatly increased. However, there has been scant data on lymph flow in the intact lymphatic system with systemic circulatory congestion. In the present study, thoracic duct and peripheral lymph flow were qualitatively determined using heated cross-thermocouples in seven mongrel dogs. Central venous pressure was raised artificially by infusing large volumes of crystalloid solution equivalent to a maximum of 30% of body weight. Although both thoracic duct and peripheral lymph flow increased with an intact (closed) lymphatic system, the increase was notably less than with a transected (opened) cervical thoracic duct. With systemic circulatory congestion, cannulated thoracic duct lymph flow circumvents a major lymph impediment to lymph flow (i.e. high central venous pressure) and therefore considerably overestimates in vivo central lymph flow in this condition.

[Ultrasonically guided puncture of the prostate and seminal vesicles].

Ultrasonically guided puncture of the prostate was carried out in 10 patients with prostatic disease. Biopsy of the aiming portion of the prostate was performed with more certainty and safety by this method than the conventional blind procedure. In the 15 cases having the suspicion of seminal vesicle disorder, the seminal vesicle was punctured under ultrasound control. By this procedure, the seminal vesicle fluid was safely and accurately obtained.

[Clinical use of an extracorporeal shock wave lithotriptor, Biolithos MARK III, for upper urinary tract stone].

The results of clinical use of an extracorporeal shock wave lithotripter (Biolithos MARK III) are presented. From May 1991 through February 1992, a total of 50 sessions were carried out on 33 patients with upper urinary tract stones. Treatments were performed on an outpatient basis, and none of the patients needed anesthesia or analgesia. One month after the last session, 18 patients (54.5%) were free from stone fragments and 6 (18.2%) had stone fragments equal to or smaller than 4 mm. The over-all successful rate obtained by these categories was 72.7%. After treatment no serious complications were observed. Although gross hematuria appeared in almost all patients, pain was noted in only 5 patients. Laboratory data after treatments showed slight and transient changes. It is concluded that Biolithos MARK III is useful and safe in the management of upper urinary tract stones on an outpatient basis.

[Two siblings with 2,8-dihydroxyadenine urolithiasis].

We treated two children with 2,8-dihydroxyadenine urolithiasis for over 7 years. The male prepositus was admitted to the hospital because of anuria when he was 10 months old. Bilateral urinary stones had caused the anuria. The stones were 2,8-dihydroxyadenine and his APRT activity was low. He has been treated with about 5.0 mg/kg/day of allopurinol without purine diet restriction. His sister, 3 years old at that time, also was found to have a renal stone. She has been treated with about 3.3 mg/kg/day of allopurinol without restricting purine. The allopurinol therapy without purine-restriction resulted in normal growth of both children with neither the recurrence of stone nor renal impairment.

[Clinical effect of allylestrenol on benign prostatic hypertrophy].

Allylestrenol at the daily dose of 50 mg was administered to 45 patients with benign prostatic hypertrophy. The treatment was performed for more than 12 weeks in 40 patients, and improvements, marked and moderate, were observed in 22 patients (55%) in the overall judgement. As side effects of this drug, decrease of potency was observed in 3 cases, decrease of libido in 1 case, pigmentation on breasts in 2 cases, palpitation in 2 cases, short breath in 1 case, and gastrointestinal symptoms in 1 case. However, these side effects were not serious. Our trial suggests that the treatment of benign prostatic hypertrophy with allylestrenol can be useful in urological clinics.

[Results of the treatment of prostatic cancer].

Two hundred and twenty patients with prostatic cancer were treated in our clinic during the past ten years between April, 1977 and March, 1987. The age distribution was from 45 to 91 years old and more than half of patients were in seventies. Stages A, B, C and D were 3.5%, 19.7%, 21.2% and 55.6%, respectively. Hormonal therapy was given in 175 cases (79.5%) as an initial treatment. The first therapy showed effectiveness in 181 (83.8%) of 216 cases; in 153 (87.4%) of 175 cases treated by hormonal therapy. Reactivation after the initial treatment was observed in 59 (32.6%) of 181 cases; in 48 (31.4%) of 153 cases treated by hormonal therapy. The interval between the start of treatment and reactivation for the stage D was significantly shorter than that for the other stages. Elevation of serum alkaline-phosphatase levels, accelerated erythrocyte sedimentation rate and hydronephrosis were significant risk factors for reactivation. Of the 220 cases, 51 (23.2%) died of advanced cancer. The overall 5-year survival rate was 41.2%. High grade and high stage were significantly related to the poor prognosis. In our studies, as hormonal therapy, maintenance on 100 mg of diethylstilbestrol diphosphate daily was found effective for the treatment of prostatic cancer.

[A case of fulminant hepatitis after reexposure to halothane six years later].

We reported a case of halothane-induced fulminant hepatitis with acute renal failure which developed 6 days after reexposure to halothane. The patient was a 58-year-old female. She had a history of liver dysfunction after exposure to halothane 6 years previously. She had surgical treatment of clubfoot under halothane anesthesia in other hospital. Preoperative physical examination and laboratory data were normal. On the 6th post-operative day she abruptly developed high fever and general fatigue. Next day, she was transferred to our hospital. At admission, fulminant hepatitis complicated with acute renal failure was diagnosed with severe liver and renal damage. She was immediately treated with plasma exchange, glucose-insulin therapy, and hemodialysis. Serum transaminase level returned to normal value within a week. However, despite repeated hemodialysis, renal function did not improve, and she died of P. aeruginosa sepsis on 28th day after the operation. It may be suggested that in this patient hypersensitivity to halothane has persisted during the six years.

Crystal structures and transistor properties of phenyl-substituted tetrathiafulvalene derivatives.

The crystal structures, thin-film properties, and field-effect transistor (FET) characteristics of tetrathiafulvalene (TTF) derivatives with two phenyl groups are systematically investigated. The highest mobility, 0.11 cm(2) V(-1) s(-1), is observed in biphenyl-substituted TTF (1). The correlation between the crystal structures and the FET properties demonstrates that good transistor properties are associated with two-dimensional intermolecular interaction, which is achieved when the molecules are standing nearly perpendicular to the substrate. Since these TTF derivatives are strong electron donors, the use of a metallic charge-transfer salt (TTF)(TCNQ) as the source and drain electrodes has resulted in a considerable reduction of the off current (TCNQ: tetracyanoquinodimethane).

Inhibition of experimental cancer cachexia by anti-cytokine and anti-cytokine-receptor therapy.

Cachexia consists of a constellation of metabolic changes that occur in cancer patients, including the reduction of muscle and fat tissue, asthenia, anorexia, hypoglycemia and hypercalcemia. These syndromes complicate therapeutic intervention and decrease the quality of life of the patient. This review discusses the involvement of cytokines in cancer cachexia and describes the contribution of IL-6 and other cytokines to the wasting of C-26-bearing mice. The neutralization of IL-6 by antibody, or IL-6 receptor antagonism by suramin, significantly reduce the severity of key parameters of cachexia. The participation of several other factors (PGE2, IL-1, IL-10 and TNF-alpha) in the cellular communication between the C-26 tumor cell and tumor-infiltrating macrophages is also described.

IL-4 and IL-13 modulate IL-10 release in endotoxin-stimulated murine peritoneal mononuclear phagocytes.

Several recent reports presented conflicting data on the action of IL-4 and IL-13 in regulating the release of proinflammatory cytokines by human monocytes. Here we show that the regulation of cytokine release by IL-4 and IL-13 could be either inhibitory or stimulatory in LPS-treated murine peritoneal macrophages. When macrophages were treated with IL-13 or IL-4, between 6 and 24 hr prior to endotoxin challenge, TNF alpha and IL-6 levels were significantly augmented. On the other hand, when the cells were cotreated with LPS plus IL-13 or IL-4, the release of TNF alpha and IL-6 was inhibited. These effects of IL-4 and IL-13 were associated with the modulation of IL-10; pretreatment resulted in a decrease, whereas cotreatment gave rise to a dramatic increase in IL-10 levels. The inhibitory effect of IL-4 and IL-13 on the release of TNF alpha was partially reversed by neutralizing anti-IL10 antibody, and the inhibition of IL-6 release was completely reversed by the antibody. These data suggest that the mechanism of action of IL-13 and IL-4 in modulating macrophage TNF alpha and IL-6 release partially involves IL-10.

The immunomodulator AS-101 inhibits IL-10 release and augments TNF alpha and IL-1 alpha release by mouse and human mononuclear phagocytes.

AS-101 is a tellurium-based compound with known immunomodulating properties. The ability of AS-101 to potentiate the effects of chemotherapeutic drugs and augment cytokine production in vivo has led to clinical trials on AS-101 which are currently being carried out in cancer patients. In the present study we show that AS-101 selectively augments the release of TNF alpha and IL-1 alpha and inhibits the release of IL-10 by lipopolysaccharide (LPS)-stimulated mouse peritoneal macrophages and human monocytes. It does not significantly affect the release of IL-6 or leukemia inhibitory factor (LIF). By itself AS-101 does not induce the release of any of these cytokines. Analysis of IL-10 and TNF alpha RNA levels using semiquantitative PCR reveals that AS-101 blocks the transcription of IL-10 mRNA, but does not significantly affect TNF alpha mRNA. Although both AS-101 and interferon (IFN)-gamma inhibit IL-10, AS-101, unlike IFN-gamma, does not prime macrophages for LPS-induced nitric oxide release and does not appear to significantly affect monocyte HLA-DR expression. Our data suggest that AS-101 is a partial IFN-gamma agonist and may explain the shift toward the release of Th-1 type cytokines observed in AS-101-treated patients.

Potential involvement of IL-10 in suppressing tumor-associated macrophages. Colon-26-derived prostaglandin E2 inhibits TNF-alpha release via a mechanism involving IL-10.

The administration of murine TNF-alpha to colon (C)-26 bearing mice, significantly protects the host against the catabolic effects of the tumor. This effect of exogenous TNF-alpha can be primarily attributed to tumor lysis rather than to a direct anticachectic action. Murine peritoneal macrophages cultured with the C-26 line or with C-26 culture supernatant do not release TNF-alpha in response to LPS stimulation. The reduction in TNF-alpha levels is associated with a significant increase in IL-10 levels. Single cell suspension of freshly disaggregated C-26 tumor (which contains host macrophages), do not produce TNF-alpha but contain significant levels of PGE2 and IL-10. In contrast, PGE2 but not TNF-alpha or IL-10 can be detected in the C-26.IVX cell line that is used to generate tumors in vivo. Neutralizing anti IL-10 Ab but not isotype-matched Ab, significantly reverses the inhibitory effect of the tumor cells and their culture supernatant on macrophage TNF-alpha release. Additional evidence is presented to indicate that the C-26-derived inhibitory activity is related to PGE2. Taken together, these results support the hypothesis that tumor-derived PGE2 prevents tumor-infiltrating macrophages from producing TNF-alpha, in part by augmenting macrophage IL-10 synthesis.

Cyclic nucleotide phosphodiesterase type IV participates in the regulation of IL-10 and in the subsequent inhibition of TNF-alpha and IL-6 release by endotoxin-stimulated macrophages.

We have recently shown that PGE2 inhibits the release of TNF-alpha from LPS-stimulated murine peritoneal macrophages via a feedback mechanism involving IL-10. Here we demonstrate that a rolipram-sensitive phosphodiesterase (PDE) type IV participates in the regulation of IL-10 synthesis. Selective PDE IV inhibitors (rolipram and RO-20-1724), but not selective inhibitors of other types of PDE, significantly augment marcrophage IL-10 production and contribute to the inhibition of TNF-alpha and IL-6 release. The addition of rolipram to LPS-stimulated macrophages results in the accumulation of cAMP and in the significant augmentation of IL-10 release. Competitive PCR analysis reveals that the drug dramatically increases IL-10 mRNA, but does not affect TNF-alpha mRNA. The inhibitory effect of rolipram on TNF-alpha can be significantly but incompletely reversed by anti-IL-10 Ab, whereas the effect of the drug on IL-6 can be completely reversed by anti-IL-10. In endotoxemic mice, the administration of rolipram increases serum IL-10 and reduces TNF-alpha and IL-6 levels. Northern blot analysis of spleens from these mice shows that rolipram increases IL-10 mRNA, whereas TNF-alpha mRNA remains largely unchanged. These results suggest that a rolipram-sensitive PDE type IV is involved in the production of IL-10 and in turn contributes to the inhibition of TNF-alpha and IL-6 release.

Vesnarinone is a selective inhibitor of macrophage TNF(alpha) release.

Vesnarinone is an experimental drug that has been used successfully in the treatment of congestive heart failure patients. In this report we investigate the effect of vesnarinone on the cytokine secretory products of mononuclear phagocytes. In a concentration-dependent manner, the drug inhibits the endotoxin(LPS)-stimulated release of tumor necrosis factor (TNF) alpha and suppresses interleukin(IL)-6 release, but does not affect the release of IL-1 alpha, IL-10 and leukemia inhibitory factor (LIF) by mouse peritoneal macrophages. Using competitive polymerase chain reaction (PCR) analyses, we find that vesnarinone significantly reduces TNF(alpha), but not IL-10 mRNA. In addition to LPS, the drug inhibits TNF(alpha) release induced by several other stimuli. The inhibitory effect of the drug on the TNF(alpha) biosynthesis can be observed in differentiated human monocytes, in macrophage cell lines, and in synovial adherent cells from rheumatoid arthritis patients. Although the precise mode of action of vesnarinone in the signal transduction pathway leading to the selective inhibition of TNF(alpha) is not known, the drug might be useful in the treatment of diseases involving that cytokine.