Morio Ohtsuka

Tissue inhibitor of metalloproteinases-2 gene polymorphisms in chronic obstructive pulmonary disease.

Proteinase/antiproteinase imbalance is the most widely accepted theory for development of chronic obstructive pulmonary disease (COPD). Mutations of tissue inhibitor of metalloproteinases-2 (TIMP-2) that downregulate its activity may increase the activities of matrix metalloproteinases and result in the degradation of the lung matrix. Polymorphisms of the TIMP-2 gene were investigated in 88 COPD patients and 40 control subjects. The variations were examined by single-strand conformational polymorphism analysis followed by sequencing. Two polymorphisms were identified, +853 GIA and -418 G/C nucleotide substitutions. There was a significant deviation in the genotypic frequencies at +853 and the allele frequencies for G were significantly higher in the COPD patient group than in the control group. For locus -418, the allele frequencies for C in the COPD patient group also tended to be higher than those in the control group. The +853 G/A nucleotide substitution was a silent variant. The -418 G/C substitution was located in the consensus sequence for the Sp1 binding site. These polymorphisms may be associated with the development of chronic obstructive pulmonary disease, decreasing the transcription and stability of the messenger ribonucleic acid, and available as genetic markers of susceptibility to the disease.

Increased levels of KL-6 and subsequent mortality in patients with interstitial lung diseases.

Objectives.  KL‐6 is a specific marker in patients with interstitial lung diseases (ILDs); however, the relationship between elevated levels of KL‐6 and subsequent mortality is not well defined. To determine if elevated serum levels of KL‐6 are associated with increased mortality, and to identify the most suitable cut‐off level of KL‐6 by which to distinguish between good prognosis and poor prognosis, we evaluated the prognostic significance of serum KL‐6 levels in patients with stable‐state ILDs.

Elimination of Neutrophils by Apoptosis During the Resolution of Acute Pulmonary Inflammation in Rats

Abstract. We evaluated the apoptosis of neutrophils during the resolution of acute pulmonary inflammation induced by exposure to ozone. The inflammatory response was assessed in rat lungs 0, 1, 3, and 7 days after 4-h exposure to air or 2 ppm ozone. Analysis of bronchoalveolar lavage fluid demonstrated significant increases in albumin concentrations on days 0 and 1 and in the number of lavageable neutrophils on days 0, 1, and 3, indicating the presence of acute pulmonary inflammation. These parameters returned to control values on day 7, which suggests that the acute pulmonary inflammation induced by ozone was reversible. On days 1 and 3, but not on day 0, the neutrophils showed morphologic evidence of apoptosis. Based on morphologic analysis, the proportion of apoptotic neutrophils was 23.3 ± 2.2% on day 1 and 55.7 ± 3.2% on day 3. Terminal deoxynucleotidyl transferase-mediated dUTP end labeling (TUNEL), in contrast, revealed that the proportion of apoptotic cells was 59.7 ± 9.1% on day 1 and 68.0 ± 4.3% on day 3. On day 3, light microscopy and electron microscopy demonstrated engulfment of the neutrophils by macrophages. These findings indicate that the apoptosis of neutrophils followed by their engulfment by macrophages contributes to the clearance of neutrophils from the sites of inflammation. Moreover, TUNEL detected apoptotic neutrophils with greater sensitivity compared with morphologic assessment.

Hypersensitivity Pneumonitis Induced by Toluene Diisocyanate: Sequelae of Continuous Exposure

A 41-year-old automobile paint sprayer showed the clinical features of hypersensitivity pneumonitis 1 week after he had begun to work with paint materials containing toluene diisocyanate. His symptoms began 6 to 8 hours after exposure to the agent and spontaneously disappeared by the next morning. He had diffuse, fine reticulonodular shadows on a chest roentgenogram and a restrictive impairment of pulmonary function. Immunoglobulin G antibody to toluene diisocyanate-human serum albumin was present in bronchoalveolar lavage fluid and sera: IgA antibody was present only in bronchoalveolar lavage fluid. Also, the patient had sensitized bronchoalveolar lymphocytes to toluene diisocyanate-human serum albumin. The histologic findings suggested hypersensitivity pneumonitis. The results of bronchoalveolar lavage, which was repeated on four separate occasions, showed lymphocytosis and a predominance of suppressor-cytotoxic T cells. The findings from serial determinations of humoral antibodies showed no changes consistent with the results of clinical and laboratory studies. In contrast, blastogenic responses of bronchoalveolar lymphocytes to toluene diisocyanate markedly decreased, and the patient showed clinical improvement despite continued exposure to the agent.

Liver metastasis at the time of initial diagnosis of lung cancer.

In order to evaluate clinicopathological features associated with liver metastases from lung cancer, we reviewed our experience of lung cancer patients seen in our division. Of the 1073 lung cancer patients diagnosed between October 1976 and May 2002, 62 (5.8%) patients had liver metastasis. The incidence of liver metastasis was 17.5% in small-cell lung cancer (SCLC) patients, whereas the incidence in non-small-cell lung cancer patients was 3.8%. Of the 62 patients, 17 had sole liver metastasis, and the remaining 45 had synchronous spread to the liver and one or more other organs. Six of 12 squamous cell carcinoma patients and 10 of 28 SCLC patients had sole liver metastasis. However, 19 of 20 adenocarcinoma patients showed liver metastasis with one or more other organs. In morphological liver metastasis, 26 of the 28 SCLC patients had multiple nodules, whereas 16 of the 34 non-small-cell lung cancer patients had a solitary liver nodule (p=0.0006). Liver is a possible site of extrathoracic spread of disease for some patients with lung cancer, especially with SCLC. When the histological types are squamous cell carcinoma or SCLC, it would also be considered likely that an isolated liver mass represents a metastasis even though there is no metastatic disease elsewhere.

Cloning of rat eotaxin: ozone inhalation increases mRNA and protein expression in lungs of brown Norway rats.

The C-C chemokine eotaxin is thought to be important in the selective recruitment of eosinophils to the site of inflammation in guinea pigs, mice, and humans. We isolated the rat eotaxin gene to determine whether a similar molecule might play a role in the pulmonary infiltration of eosinophils during acute inflammation in the rat. The cDNA for rat eotaxin encoded a 97-amino acid protein containing a 74-amino acid mature eotaxin protein with 97.3% identity to mouse eotaxin. The recombinant protein encoded by this gene displayed specific chemotactic activity for eosinophils when analyzed with a microchemotactic chamber. The expression of eotaxin mRNA increased approximately 1.6-fold immediately after exposure to ozone and was 4-fold higher after 20 h. The number of lavageable eosinophils at the same time points were 3- and 15-fold greater, respectively, than control eosinophils. Immunocytochemistry revealed that alveolar macrophages and bronchial epithelial cells were positive for eotaxin. These results suggest that eotaxin may be involved in the recruitment of eosinophils into the air spaces during certain inflammatory conditions in rats.The C-C chemokine eotaxin is thought to be important in the selective recruitment of eosinophils to the site of inflammation in guinea pigs, mice, and humans. We isolated the rat eotaxin gene to determine whether a similar molecule might play a role in the pulmonary infiltration of eosinophils during acute inflammation in the rat. The cDNA for rat eotaxin encoded a 97-amino acid protein containing a 74-amino acid mature eotaxin protein with 97.3% identity to mouse eotaxin. The recombinant protein encoded by this gene displayed specific chemotactic activity for eosinophils when analyzed with a microchemotactic chamber. The expression of eotaxin mRNA increased ∼1.6-fold immediately after exposure to ozone and was 4-fold higher after 20 h. The number of lavageable eosinophils at the same time points were 3- and 15-fold greater, respectively, than control eosinophils. Immunocytochemistry revealed that alveolar macrophages and bronchial epithelial cells were positive for eotaxin. These results suggest that eotaxin may be involved in the recruitment of eosinophils into the air spaces during certain inflammatory conditions in rats.

Analysis of Bronchoalveolar Lavage Cells and Fluids in Patients with Hypersensitivity Pneumonitis: Possible Role of Chemotactic Factors in the Pathogenesis of Disease

The current study concerns immunological mechanisms involved in the pathogenesis of hypersensitivity pneumonitis (HP) by an analysis of the cells and chemotactic factors (CF) obtained by bronchoalveolar lavage (BAL). Nine patients at an acute stage (HP acute, 8 summer type and 1 pigeon breeders lung) and 4 patients at a quiescent stage (HP quiescent, 3 summer type and 1 pigeon breeders lung) were included. The results indicate that: CF for polymorphonuclear cells (PMN) in HP acute were significantly more potent than in HP quiescent; CF for mononuclear cells were not significantly different in the groups; the percentage of lymphocytes in HP acute was significantly greater even though HP quiescent revealed greater percentages of lymphocytes as compared to normal controls; determination of T cell subsets employing OKT antibodies revealed the ratio of OKT8+ cells to OKT4+ cells in HP acute was significantly higher than in HP quiescent, and chemotaxis for PMN was marginally correlated with the percentage of OKT8+ cells at the acute stage of disease.

Conserved CDR 3 region of T cell receptor BV gene in lymphocytes from bronchoalveolar lavage fluid of patients with idiopathic pulmonary fibrosis

Idiopathic pulmonary fibrosis (IPF) is an inflammatory lung disease characterized by the accumulation of inflammatory cells and deposition of collagen, resulting in lung remodelling. High numbers of T cells are present in bronchoalveolar lavage fluid (BALF) of IPF patients, although the characteristics of these cells are yet to be determined. To elucidate the pathogenic mechanisms of IPF, we analysed the T cell receptor (TCR) of BALF lymphocytes in three patients with IPF and three healthy subjects as control. TCR repertoire of BALF lymphocytes and T cell clonality were examined by family PCR and Southern blot analysis, and single‐strand conformation polymorphism (SSCP), respectively. We observed that the TCR repertoire in the lung was heterogeneous, both in the control subjects and three patients with IPF. SSCP analysis demonstrated an increase in the number of accumulated T cell clones in BALF of two of the three patients, but not in the healthy subject. Furthermore, junctional sequence analysis showed the presence of conserved amino acid motifs (ETGRSG, LAxG, QGQ, GxQP, GRxG, VAR, PGT, GTI, GGT, TGR, LxLxQ, SGQ) in the TCR‐CDR 3 region of BAL lymphocytes in patients with IPF, whereas only two amino acid motifs (VTTG, GGE) were found in the control. Our findings suggest that T cells in BALF of patients with IPF expand oligoclonally in the lung, suggesting antigen stimulation of these cells.

Simultaneous measurements of KL-6 and SP-D in patients undergoing thoracic radiotherapy.

PurposeRadiation pneumonitis (RP) is a serious complication in patients undergoing thoracic radiotherapy (TRT). Serum KL-6 and SP-D have been shown to increase in several kinds of interstitial pneumonia. To evaluate their clinical usefulness in detecting RP, we serially measured them in patients receiving TRT.Materials and MethodsThirty-nine patients, who received TRT for lung cancer between July 1999 and April 2004, were prospectively studied. Serum levels of KL-6 and SP-D were measured using enzyme-linked immunosorbent assays. Patients were followed up until August 2004 or their deaths.ResultsDuring the period, RP occurred in 19 patients. In five patients with diffust RP extended outside the radiation field, serum KL-6 levels increased, reaching more than 1000 U/mL. Serum KL-6 levels at 40 gy in patients who developed RP were higher than those without it (p=0.0363, Mann-Whitney U test). In addition, serum KL-6 levels at 40 Gy in patients who developed RP were higher than those of pretreatment (p=0.0126, Wilcoxon signed rank test). On the other hand, there were no statistical differences between SP-D at 40 Gy and those before TRT (p=0.1165).ConclusionsIncreased KL-6 at 40 Gy compared with those before treatment in patients undergoing TRT may be of clinical significance. KL-6 proved to be a useful indicator for estimating RP, while usefulness of SP-D was not confirmed in this study.

Thymic and pulmonary mucosa-associated lymphoid tissue lymphomas in a patient with Sjögren’s syndrome and literature review

Abstract:  Mucosa‐associated lymphoid tissue lymphoma (MALToma) has been reported in several organs. Among MALTomas, thymic and pulmonary MALTomas are rare. The present report describes a patient with Sjögren’s syndrome who presented thymic and pulmonary MALTomas. Although the exact pathogenetic relationship between these two tumours is uncertain, it is likely that the underlying immune dysregulation related to Sjögren’s syndrome contributed to the occurrence and the unusual manifestation of MALTomas in this patient.