Yasuharu Abe

The cytokines NAP-1 (IL-8), MCP-1, IL-1 beta, and GRO in rabbit inflammatory skin lesions produced by the chemical irritant sulfur mustard

Developing and healing dermal inflammatory lesions were produced in rabbits by the topical application of dilute sulfur mustard (SM),9 the military vesicant. In tissue sections of such lesions, cells containing the mRNA of important cytokines were identified with in situ hydridization techniques. These cytokines were neutrophil attractant/activation protein-1 (NAP-1 (also called IL-8)), monocyte chemoattractant (activating) protein 1 (MCP-1), interleukin 1 (beta) (IL-1 (beta)), and GRO (a growth factor and chemokine). Mononuclear cells (mainly macrophages and activated fibroblasts) contained the mRNA of all four of these cytokines. A higher percentage of cytokine-producing mononuclear cells (macrophages and activated fibroblasts) was present in lesions at 2 days (their peak size) than at 6 days, when they were almost healed. Granulocytes emigrated from the bloodstream, passed through the lesions, and were the major constituent of the protective crust. This sequence correlated with the distribution of cells able to produce NAP-1: At 2 days and 6 days, the mononuclears that contained messenger RNA for this granulocyte chemoattractant were found mainly in the upper part of the dermis. At 2 days and 6 days, cells containing the mRNA of IL-1, a primary cytokine, were also found predominantly in the upper dermis, i.e., nearest the site of injury. In contrast, mononuclears containing the mRNA of MCP-1 (a monocyte chemoattractant), and the mRNA of GRO (a granulocyte chemoattractant) were more equally distributed throughout the dermis. SM stimulated hair follicle epithelial cells to up-regulate GRO mRNA and, to a lesser degree, NAP-1 mRNA. Apparently, the irritation produced by SM directly or indirectly induces such epithelial cells to manufacture these growth factors. In the rabbit, hair follicles are known to be the main source of new epithelial cells after the covering epithelium has been destroyed. Therefore, GRO is probably a major autocrine-paracrine stimulus for such repair. A brief review of the role of cytokines in dermal inflammation is presented.

A case of malignant pleural mesothelioma following exposure to atomic radiation in Nagasaki

Abstract We report the case of a 75‐year‐old Japanese man who developed malignant mesothelioma in the left hemithorax 50 years after the dropping of the atomic bomb on Nagasaki in 1945. This may be the first reported case of malignant mesothelioma following exposure to atomic radiation. Asbestos is the leading cause of malignant mesothelioma, but radiation therapy is the primary non‐asbestos‐related cause. In the case of radiation therapy, the interval between exposure and the occurrence of malignant mesothelioma tends to be many years. This patient was at a high risk of malignant mesothelioma as he had been exposed to radiation from the atomic bomb and may also have had a history of asbestos exposure at the munitions factory where he was employed as a shipbuilder for 2 years. It has been suggested that combined exposure to atomic radiation and asbestos is associated with an increased incidence of malignant mesothelioma. If thickening of the pleura or pleural effusion is found in atomic bomb survivors, malignant mesothelioma should be considered as one of the options in the differential diagnosis, even although the atomic bomb attacks occurred several decades ago.

In vitro angiotensin-converting enzyme and interleukin-1 production by epithelioid cells isolated from induced rabbit lung granuloma

We attempted in this experiment to culture epithelioid cells isolated from granuloma induced in the lung of female rabbits that had been injected intravenously with Freunds complete adjuvant. The morphologic findings of isolated cells included abundant rough-surfaced endoplasmic reticulum, mitochondria, Golgi complexes, electron-dense lysosomes, and numerous cytoplasmic processes on the cell surface. Enzymatically these cells were positive for acid alpha-naphthyl acetate and beta-galactosidase staining. These findings coincided with the expected morphologic and enzymatic characteristics of epithelioid cells examined in vivo. Cells isolated from this granuloma are thought to be more than 86% epithelioid cells. Additionally, 14.4 +/- 4.3% of the epithelioid cells showed phagocytosis of latex beads, low when compared with the 83.9 +/- 5.2% value of alveolar macrophages obtained from rabbits injected with adjuvant 4 weeks before sacrifice. The isolated epithelioid cells were incubated for 24 h without decrease in cell population. Their culture supernatants showed 1.86 +/- 0.38 units/ml of angiotensin-converting enzyme (ACE) activity, which was inhibited by cycloheximide. The culture supernatants also showed interleukin-1 (IL-1) activity levels of 6411 +/- 914 cpm without lipopolysaccharide (LPS) stimulation. This increased to 21,766 +/- 3026 cpm with LPS stimulation. In view of these results, we believe it is possible to incubate epithelioid cells for up to 24 h during which time they will produce ACE and IL-1 in the culture supernatants.

Identification of Il-1 Inhibitory Activity as Il-1 Receptor Antagonist (Il-1Ra) in Vitro and Kinetics of Il-1RA and Il-1Bt in Experimental Rabbit Lung Granuloma

This study investigated whether interleukin-1 (IL-1) inhibitory activity in LPS-stimulated culture supernatants of alveolar macrophages and epithelioid cells obtained from rabbit lung granulomas induced by complete Freunds adjuvant (CFA) was identical to IL-1 receptor antagonist (IL-1ra) and examined the changes of IL-1ra and IL-1 beta levels in lung tissue during the natural course of granulomatous inflammation. In the thymocyte proliferation assay, prostaglandin E2 (PGE2)-free culture supernatants from each cell population revealed a bell-shaped IL-1 titration curve with IL-1 activity suppressed at dilutions of 1:1 to 1:2, and gel chromatography of serum-free culture supernatants showed an IL-1 inhibitory peak at 21-25 kD. Suppression of IL-1 activity in the supernatants at lower dilutions and the gel-purified IL-1 inhibitory activity both almost disappeared after IL-1ra depletion with an anti-rabbit IL-1ra immunoaffinity column, indicating that IL-1ra was responsible for this in vitro IL-1 inhibitory activity. Pulmonary tissue levels of IL-1 beta pecked at 24 h (52.0 +/- 9.5 pg/mg) after CFA injection, whereas IL-1ra levels peaked at 4 weeks (23.1 +/- 4.0 ng/mg) when granuloma development was maximal, and the molar excess of IL-1ra to IL-1 beta peaked from 4 weeks onward at over 800-fold. These observations suggest that IL-1ra may be effective for IL-1 regulation, especially in the later stage of granulomatous responses. Immunohistochemical studies demonstrated that the cellular source of IL-1ra within the pulmonary granulomas was mainly epithelioid cells.

α-Naphthyl Acetate Esterase-1 (ANAE-1) Secreted by Epithelioid Cells from Induced Rabbit Lung Granuloma Showed MIF Activity

The function of alpha-naphthyl acetate esterase-1, whose isoelectric point values range from 5.15 to 5.45, was examined. A higher value of alpha-naphthyl acetate esterase-1 was detected in the extracts of epithelioid cells isolated from rabbit lung granuloma at 4 weeks after injection of Freunds complete adjuvant, compared to those values of alveolar macrophages isolated from the same lungs described above and of the normal lungs. Additionally, this enzyme activity was observed to be prominent in the culture supernatants of epithelioid cells. alpha-Naphthyl acetate esterase-1 was purified from lung granuloma as a single 62-kDa band by SDS-PAGE analysis. The purified enzyme showed a macrophage migration inhibition activity at concentrations over 20 nM, and its activity was dose-dependent. Moreover, when various amounts of the purified enzyme were added to lymphocyte-derived macrophage migration inhibitory factor, macrophage migration inhibition was significantly enhanced with a dose-dependent manner. The results suggest that alpha-naphthyl acetate esterase-1 secreted by granuloma macrophages, particularly by epithelioid cells, contributes to granuloma formation.