Kyong-Yob Min

Activation of lavage lymphocytes in lung injuries caused by radiotherapy for lung cancer

PURPOSE Radiation pneumonitis sometimes extends beyond the irradiated area of a lung and can also affect the opposite lung. Some immunological mechanisms, in addition to simple direct injury of the lungs by radiation, seem to be involved in the onset of radiation pneumonitis. To clarify such mechanisms, the effects of radiation on local inflammatory cells in lungs, in particular, lymphocytes, were examined. METHODS AND MATERIALS A comparison was made of bronchoalveolar lavage fluid (BALF) findings from 13 irradiated patients (RT group) and 15 nonirradiated patients (non-RT group) with lung cancer. Patients who later developed radiation pneumonitis (RP group) and those who did not (RP-free group) were also compared. Using a two-color flowcytometer, radiation-induced changes in local inflammatory cells in lungs were analyzed. This included analyses of human leukocyte-associated antigen (HLADR) and intercellular adhesion molecule-1 (ICAM-1) expression on T-cells, which are though to be involved in cell activation and interactions between cells. RESULTS The following aspects of BALF were higher in the RT group than in the non-RT group: (a) the percentage of lymphocytes and eosinophiles; (b) the incidence of HLADR-positive CD4+T-cells and HLADR-positive CD8+T-cells; and (c) the incidence of ICAM-1--positive T-cells. The following aspects of BALF were higher in the RP group than in the RP-free group: (a) the total cell counts; (b) the percentage of lymphocytes; and (c) the incidence of ICAM-1-positive T-cells. A significant relationship was seen between the incidence of ICAM-1 expression on T-cells and the number of days from the initiation of radiotherapy to the onset of radiation pneumonitis. CONCLUSION These data suggest that irradiation can induce accumulation of activated T-cells (HLADR and ICAM-1--positive T-cells) in the lung. This accumulation may be closely linked to radiation-induced lung injury. It is also suggested that the incidence of ICAM-1--positive T-cells in BALF may serve as a useful clinical marker of radiation pneumonitis.

β2‐adrenergic regulation of ciliary beat frequency in rat bronchiolar epithelium: potentiation by isosmotic cell shrinkage

Single bronchiolar ciliary cells were isolated from rat lungs. The β2‐adrenergic regulation of ciliary beat frequency (CBF) was studied using video‐optical microscopy. Terbutaline (a β2‐adrenergic agonist) increased CBF in a dose‐dependent manner, and it also decreased the volume of the ciliary cells. These terbutaline actions were inhibited by a PKA inhibitor (H‐89) and mimicked by forskolin, IBMX and DBcAMP. Ion transport inhibitors were used to isosmotically manipulate the volume of the terbutaline‐stimulated bronchiolar ciliary cells. Amiloride (1 μm) and bumetanide (20 μm) potentiated cell shrinkage and the CBF increase, and they shifted the terbutaline dose–response curve to the lower‐concentration side. Quinidine (500 μm), in contrast, increased cell volume and suppressed the CBF increase. Moreover, a KCl solution containing amiloride (1 μm) and strophanthidin (100 μm) increased cell volume and suppressed the CBF increase, and then the subsequent removal of either amiloride or strophanthidin decreased cell volume and further increased CBF. NPPB (10 μm) or glybenclamide (200 μm) had no effect on the action of terbutaline. Thus, in terbutaline‐stimulated ciliary cells, cell shrinkage enhances the CBF increase; in contrast, cell swelling suppresses it. However, the results of direct manupulation of cell volume by applying osmotic stresses (hyperosmotic shrinkage or hyposmotic swelling) were the opposite of the findings of the isosmotic experiments: hyposmotic cell swelling enhanced the CBF increase, while isosmotic swelling suppressed it. These results suggest that isosmotic and non‐isosmotic volume changes in terbutaline‐stimulated bronchiolar ciliary cells may trigger different signalling pathways. In conclusion, terbutaline increases CBF and decreases the volume of rat bronchiolar ciliary cells via cAMP accumulation under isosmotic conditions, and the isosmotic cell shrinkage enhances the CBF increase by increasing cAMP sensitivity.

Segmental differences in vasodilatation due to basal NO release in in vivo cat pulmonary vessels

This study was conducted to examine segmental differences in vasodilatation caused by the basal release of nitric oxide (NO) in the serially connected pulmonary vessels and to estimate the relative contributions of endothelial and neuronal NO synthase (NOS), and inducible NOS to the vasodilatation. Using an X-ray TV system on in vivo cat lungs, we measured internal diameter (ID) changes in resistance (100-400 microm ID), small conduit (600-1000 microm) and large conduit (1200-1700 microm) arteries, and veins of the same size. Non-selective NOS inhibitors, L-NAME (30-50 mg/kg i.v.) and L-NMMA (40-60 mg/kg i.v.), decreased the ID of all vessels studied, although their D-isomers had no effect. The decrease was larger in conduit arteries than in resistance arteries, with maximum response of small conduit arteries (25 +/- 2%), while venous segments displayed relatively uniform response (10-12%). L-Arginine completely abolished the ID decrease but hexamethonium bromide and phentolamine had no effect. Selective inhibitors of inducible NOS, L-canavanine (100 mg/kg i.v.) and S-methylisothiourea (10 mg/kg i.v.) did not affect any of the vessels. The data suggest that basal release of NO chiefly derived from endothelial NOS serves to dilate cat pulmonary arteries and veins, particularly small conduit arteries.

Cell shrinkage evoked by Ca2+‐free solution in rat alveolar type II cells: Ca2+ regulation of Na+–H+ exchange

The effects of intracellular Ca2+ concentration, [Ca2+]i, on the volume of rat alveolar type II cells (AT‐II cells) were examined. Perfusion with a Ca2+‐free solution induced shrinkage of the AT‐II cell volume in the absence or presence of amiloride (1 μm, an inhibitor of Na+ channels); however, it did not in the presence of 5‐(N‐methyl‐N‐isobutyl)‐amiloride (MIA, an inhibitor of Na+–H+ exchange). MIA decreased the volume of AT‐II cells. Inhibitors of Cl−–HCO3− exchange, 4,4′‐diisothiocyanostilbene‐2,2′‐disulfonic acid (DIDS) and 4‐acetamido‐4′‐isothiocyanatostilbene‐2,2′‐disulfonic acid (SITS) also decreased the volume of AT‐II cells. This indicates that the cell shrinkage induced by a Ca2+‐free solution is caused by a decrease in NaCl influx via Na+–H+ exchange and Cl−–HCO3− exchange. Addition of ionomycin (1 μm), in contrast, induced cell swelling when AT‐II cells were pretreated with quinine and amiloride. This swelling of the AT‐II cells is not detected in the presence of MIA. Intracellular pH (pHi) measurements demonstrated that the Ca2+‐free solution or MIA decreases pHi, and that ionomycin increases it. Ionomycin stimulated the pHi recovery after an acid loading (NH4+ pulse method), which was not noted in MIA‐treated AT‐II cells. Ionomycin increased [Ca2+]i in fura‐2‐loaded AT‐II cells. In conclusion, the Na+–H+ exchange activities of AT‐II cells, which maintain the volume and pHi, are regulated by [Ca2+]i.

Delayed shrinkage triggered by the Na+–K+ pump in terbutaline‐stimulated rat alveolar type II cells

Terbutaline (10 μm) induced a triphasic volume change in alveolar type II (AT‐II) cells: an initial shrinkage (initial phase) followed by cell swelling (second phase) and a gradual shrinkage (third phase). The present study demonstrated that the initial and the third phases are evoked by the activation of K+ and Cl− channels and the second phase is evoked by the activation of Na+ and Cl− channels. Ouabain blocked the third phase, although it did not block the initial and second phases. This suggests that the third phase is triggered by the Na+–K+ pump. Tetraethylammonium (TEA, a K+ channel blocker) decreased the volume of AT‐II cells and enhanced the terbutaline‐stimulated third phase, although quinidine, another K+ channel blocker, increased the volume of AT‐II cells. The TEA‐induced cell shrinkage was inhibited by ouabain, suggesting that TEA increases Na+–K+ pump activity. Ba2+, 2,3‐diaminopyridine and a high [K+]o (30 mm) similarly decreased the volume of AT‐II cells. These findings suggest that depolarization induced by TEA increases Na+–K+ pump activity, which increases [K+]i. This [K+]i increase, in turn, hyperpolarizes membrane potential. Valinomycin (a K+ ionophore), which induces hyperpolarization, decreased the volume of AT‐II cells and enhanced the third phase in these cells. In conclusion, in terbutaline‐stimulated AT‐II cells, an increase in Na+–K+ pump activity hyperpolarizes the membrane potential and triggers the third phase by switching net ion transport from NaCl entry to KCl release.

Multiple Cytokines-Producing Pleomorphic Carcinoma of Lung with Metastasis to the Small Intestine

A 58-year-old man underwent upper lobectomy for primary pleomorphic carcinoma of the lung. Nine months later, the pleomorphic carcinoma was recurred with marked peripheral leukocytosis and an elevated C-reactive protein. Chest and abdominal computed tomography (CT) revealed enlarged mediastinal lymph nodes and a bulky tumor in the small intestine. An enterectomy was performed and the intestinal tumor was removed. Immunostaining revealed tumor cells positive for G-CSF and TNF-α as well as an increased level of serum G-CSF and TNF-α. We describe a rare case of G-CSF and TNF-α producing pleomorphic carcinoma of the lung with metastasis to the small intestine.

Pleural vasculitides of microscopic polyangiitis with asbestos-related plaques

A 69‐year‐old man who had been exposed to asbestos for approximately 40 years presented with the complaint of fever and pleuritic chest pain on the right side on deep inspiration. Chest X‐ray films showed pleural effusion in the right side. Initial antibiotic treatment was ineffective. The hyaluronic acid level was high in the pleural effusion but no malignant mesotheliomal cells were seen with blind pleural biopsy. Blood chemistry showed a remarkable high titer of myeloperoxidase anti‐neutrophil cytoplasmic antibody (MPO‐ANCA) and open renal biopsy suggested crescentic glomerulonephritis. The precise pathological examination on the pleura obtained by the open pleural biopsy showed vasculitides and plaque leading to diagnosis of microscopic polyangiitis (MPA). This is a rare case of MPA seen in the pleural arteries.

Waldenström's macroglobulinemia: report of an autopsy case presenting with a pulmonary manifestation

An autopsy case of Waldenströms macroglobulinemia is reported, in whom an abnormal pulmonary shadow had already existed 2 years before the diagnosis of the disease and was proved to be pulmonary involvement. Immunoelectrophoresis demonstrated a monoclonal increase in immunoglobulin M with kappa light chain. A chest X-ray film showed a reticulo-nodular shadow in the right lower lobe of the lung. A bronchial biopsy specimen revealed a diffuse and dense lymphocytic infiltration. Bone marrow aspirate revealed no remarkable change except for a slight increase in plasma cells (1.7%) and an appearance of atypical lymphocytes (0.5%). At autopsy, more than half of the right lower lobe of the lung was occupied by a pale whitish, viscid and glossy tumour mass. Heptosplenomegaly and lymph node enlargement were not observed. Histological findings of the tumour tissue were similar to those of the biopsy specimen. Lymphocytic infiltration was observed also in the liver, kidneys, spleen, bone marrow and lymph nodes, but was of minor degree. Other reported cases of Waldenströms macroglobulinemia accompanied by pulmonary involvement are reviewed.