Katsuro Yagawa

Lack of Response of Murine Peritoneal Macrophages to in vitro Activation by Muramyl Dipeptide (MDP)

Peritoneal exudate macrophages from mice, rats, and guinea pigs were assessed using six different parameters of macrophage activation to determine whether the cells were stimulated under similar experimental conditions. Peritoneal exudate macrophages from mice, irrespective of strain, were far less responsive to a variety of in vitro stimulatory effects of N‐acetylmuramyl‐L‐alanyl‐D‐isoglutamine than those from rats or guinea pigs, while no significant differences were noted with the reactivity to stimulation by endotoxic lipopolysaccharide. We conclude that macrophage activation by MDP in vitro is species dependent.

Extracellular ATP itself elicits superoxide generation in guinea pig peritoneal macrophages

Extracellular ATP itself elicited the generation of superoxide (O− 2) in guinea pig peritoneal macrophages associated with an increase in cytosolic calcium ([Ca2+]). The ATP‐Induced O− 2 generation was completely inhibited by pretreatment with pertussis toxin (PT) accompanied by the suppression of (Ca2+), mobilization. Pre‐exposure to a small amount of phorbol myristate acetate (PMA) primed the ATP‐induced generation of O− 2 without it change of (Ca2+). The results suggest that ATP‐induced O− 2 generation is mediated by (Ca2+), mobilization and by PT‐sensitive G protein.

Influence of chemotherapy on superoxide anion-generating activity of polymorphonuclear leukocytes in patients with lung cancer

The superoxide anion (O  2− ) generating activity of polymorphonuclear leukocytes (PMN) in peripheral venous blood obtained from 15 patients with lung cancer was measured weekly after drug treatment. The amount of O  2− generated by the cells before chemotherapy was 5.74 ± 0.42 nmol/minute/106 cells (mean ± SE). The value decreased gradually and reached a nadir of 1.98 ± 0.24 at 2 weeks after chemotherapy, at which time PMN count also reached a nadir of 1359 ± 251/μl (mean ± SE). The decreased O  2− generating activity of the PMN recovered gradually thereafter and reached the level seen before the chemotherapy at 4 weeks. Expression of Fc receptors on the plasma membrane in PMN was measured after chemotherapy by the binding of iodine 125 (125I)‐immune complexes to the cells. Almost 40% decrease of the binding was observed in the cells obtained at 2 weeks after chemotherapy, compared with that in the cells before chemotherapy. These results indicate that PMN present in peripheral blood at 2 weeks after chemotherapy are not fully matured, in terms of O  2− generating activity and the expression of Fc receptors.

Species Dependency of In Vitro Macrophage Activation by Bacterial Peptidoglycans

The effect of various bacterial cell wall components on in vitro biological function of murine peritoneal exudate macrophages was evaluated. We examined four different parameters of metabolic activity and monokine secretion. Peritoneal exudate macrophages from rats and guinea pigs, all of the strains tested, were stimulated by whole bacterial cell wall preparations, purified bacterial cell wall peptidoglucans, its water‐soluble peptidoglycan fragments, muramyl dipeptides and amphipathic substances. Murine peritoneal exudate macrophages were activated by amphipathic substances of gram‐positive bacteria. However, macrophages from mice, irrespective of strains, were not stimulated in the in vitro assay systems by purified bacterial cell wall peptidoglycan, water‐soluble bacterial peptidoglycan fragments or muramyl dipeptides. These results suggest that macrophage activation by bacterial peptidoglycan in vitro is animal species specific.

Plasma Thrombosis-Inducing Activity in 120 Patients with Primary Lung Cancer

One hundred and twenty patients with primary lung cancer were examined for the presence of thrombosis-inducing activity (TIA) in their plasma. TIA was identified in plasma from 16 of 38 patients with stage 3 (42%) and 31 of 65 patients with stage 4 (48%) disease. On the other hand, only 1 of 17 patients with stages 1 and 2 (6%) showed TIA in their plasma. Cell type did not seem to correlate with the presence of plasma TIA, since TIA was identified in plasma from patients with all cell types. Survival of 32 patients with inoperable non-small cell lung cancer, all stage 4, was studied. The mean survival time was 7.2 months in the TIA-positive group and 10.3 months in the TIA-negative group. This difference was statistically significant.

Thrombosis-Inducing Activity in Plasma of Patients with Acute Respiratory Tract Infection Disappears after Treatment

Thrombosis-inducing activity (TIA) was identified in plasma from 16 of 27 patients (59%) with acute respiratory tract infections. On the other hand, it was present in only 9 of 79 subjects (11%) with chronic lung diseases and 4 of 49 healthy volunteers (8%). In the patients with acute respiratory tract infections, there were significant elevations in plasma fibrinogen, C-reactive protein and erythrocyte sedimentation rate in the TIA-positive group compared with the negative group. Plasma TIA disappeared in all of the 8 patients who were retested for TIA 2-5 weeks after they became disease free. Pneumonia was induced in rabbits by transbronchial injection of viable Escherichia coli. TIA was not present in plasma from normal rabbits, but it appeared in plasma collected 3 days after injection. It then disappeared after 1-2 weeks of treatment with antibiotics. TIA may serve as a marker for inflammatory responses and be a factor responsible for elevated blood coagulation activity in patients with acute infectious diseases.

Herpes Zoster in Patients with Sarcoidosis

108 patients with sarcoidosis were retrospectively studied for the development of herpes zoster. Five of these patients (4.6%), 2 of whom were in their twenties, developed herpes zoster. Only 1 patient had been treated with an oral steroid. All 5 had extrathoracic lesions. Zoster tended to occur during the inactive stage of sarcoidosis and did not exacerbate the activity of the sarcoidosis. The clinical course of their zoster infection was typically benign. There have been few reports of herpes zoster in patients with sarcoidosis. Further studies are required to determine whether sarcoidosis predisposes to herpes zoster infection.

Superoxide anion generation by polymorphonuclear leukocytes enhanced in a patient with colony-stimulating activity-producing lung cancer

We examined superoxide (O-2)-generating activity of polymorphonuclear leukocytes (PMN) from a patient with lung cancer in whom there was a marked granulocytosis. There was a high level of colony-stimulating activity (CSA) in both the serum and the supernatant of the culture of the primary tumor. The PMN from the patient produced almost three times more O-2 than did the PMN from healthy donors and other patients with lung cancers but with no granulocytosis. The binding of wheat germ agglutinin (WGA), used as a stimulus for O-2 generation, to the surface membrane of cells and the protein content in the cells were about double that noted in the case of healthy donors. Following total extirpation of the tumor, the level of CSA in the serum decreased, as did activity of PMN for O-2 generation, and the binding of WGA to the cells reverted to normal levels.

Transbronchial needle aspiration in the diagnosis of pulmonary sclerosing haemangioma

A 5 l-year-old housewife was referred to our section for the evaluation of an abnormal chest roentgenogram showing a solitary round mass shadow in the left lower peripheral lung field. The mass was characterized as a well circumscribed 22-cm homogeneous nodule without notching. There were no malignant signs on chest tomography and computed tomographic scan. No abnormalities were revealed by physical and haematobiochemical examinations. Inspection of the airways by bronchoscopy yielded normal results. Under fluoroscopic guidance, transbronchial biopsy forceps and a bronchial brush could not pass into the mass. TBNA was performed using a retractable 21-gauge needle. Cytologic smears stained by the Papanicolaou technique revealed hyperplastic alveolar epithelial cells of a somewhat dysplastic nature showing glandular cell arrangement [Plate l(a)] and macrophages, some of which had haemosiderin pigments [Plate l(b)]. Polynucleated giant cells, fibroblasts and endothelial cells were also seen. Cytologic material obtained by TBNA did not show the full range of features of SH, however, we could make the diagnosis of this entity interpreting cytologic features combined with the chest radiograph in a middle-aged woman. As the patient had an operation for colon cancer last year, she underwent left lower

Effect of transmethylation-reaction and increased levels of cAMP on superoxide generation of guinea-pig macrophages induced with wheat germ agglutinin and phorbor myristate

Superoxide (O2 −) generation of guinea‐pig macrophages induced by wheat germ agglutinin (WGA) was suppressed to a great extent by the inhibition of transmethylation with 3′‐deazaadenosine. When macrophages were stimulated with phorbor myristate (PMA) instead of WGA, the suppression of O2 − generation of macrophages was observed to be slight despite the presence of 3′‐deazaadenosine. These results were confirmed under various conditions. Thus the WGA‐stimulated O2 − generation of macrophages is probably associated with transmethylation, but the PMA‐stimulated O2 − generation is not. WGA‐stimulated O2 − generation of macrophages was also inhibited in the presence of dibutyryl cAMP or prostaglandin E2 (PGE2), substances that increase intracellular cAMP, but PMA‐stimulated O2 − generation was only slightly affected by these compounds. These results suggest that the mechanism for O2 − generation of macrophages caused by WGA is different from that for O2 − generation caused by PMA.