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We investigated the effect of interaction between lung fibroblasts and macrophages on macrophage inflammatory protein 1alpha (MIP-1alpha) production by macrophages. In a co-culture system consisting of WI-38 lung fibroblasts layered over THP-1 macrophages stimulated with lipopolysaccharide (LPS), MIP-1alpha production by THP-1 was significantly lower in co-culture with WI-38 than in THP-1 alone. Treatment with conditioned medium generated from WI-38 (CM-WI-38) suppressed MIP-1alpha production and mRNA expression in THP-1 cells. Such inhibitory effect of CM-WI-38 on MIP-1alpha production was abrogated by treatment with indomethacin, NS-398 (a specific COX-2 inhibitor), or anti-prostaglandin E(2) antibody. Furthermore, even in a transwell filter system separating both types of cells, co-culture-induced reduction of MIP-1alpha production was observed. Therefore, soluble factors such as prostaglandin E(2) released from lung fibroblasts are responsible for the co-culture-induced inhibition of macrophage-derived MIP-1alpha production, suggesting that immune and inflammatory cell interactions can contribute to the modulatory mechanisms involved in the regulation of the inflammatory or fibrotic process.

We investigated the expression of Toll-like receptors (TLRs) and associated signaling molecules in inflammatory stimuli-activated murine primary alveolar macrophage (AM) in vitro, and in a murine model of acute lung injury. The results demonstrated three patterns of gene expression: the TLR2 and myeloid differentiation factor 88 (MyD88) gene expressions were induced in AM in response to lipopolysaccharide (LPS), interleukin (IL)-1beta, or tumor necrosis factor-alpha or in the lung tissue of an LPS-induced acute lung injury model; the gene expressions of TLR1, -3, -6, CD14, and MD2 were unchanged; and the TLR4 and TLR5 gene expressions were downregulated in AM following inflammatory stimuli. Furthermore, the LPS-induced upregulation of the TLR2 gene was mediated via the activation of NF-kappaB. These results indicate that the TLR2 expression upregulated in AM following bacterial respiratory infections may render AM responsive to TLR2 ligands, which may enhance innate immunity against pathogens in the lung.

Since the ability of alveolar epithelial cells to ingest inhaled fine particles has not been characterized in detail, the present study seeks to evaluate this physiological activity. We used a 0.2% suspension of intact or lecithin-coated polystyrene latex beads (240 nm in diameter). A 5-ml suspension of intact or lecithin-coated latex beads was intratracheally administered to rats using a compressor nebulizer. Thereafter, the lungs were perfused intratracheally with glutaraldehyde solution and cut into small pieces. The samples were postfixed with osmium tetroxide, embedded in epoxy resin and examined under an electron microscope. Both lecithin-coated and uncoated beads were incorporated into alveolar macrophages. Some of the ingested beads in the alveolar macrophages were sequestered within lysosomes. Types I and II alveolar epithelial cells selectively incorporated only lecithin-coated beads, which were also observed within the cytoplasm of monocytes in the capillary lumen. These findings suggest that alveolar epithelial cells can incorporate exogenous particles, which are then transferred from the alveoli to intravascular spaces by transcytosis.

This is a report of a patient with occupational hypersensitivity pneumonitis (HP) caused by Konjak flour and powdered Hijikia fusiforme. A 56-year-old man had worked as a Konnyaku manufacturer for 38 years, and suffered from dyspnea on exertion. Konnyaku is a paste made from Konjak flour, powdered Hijikia fusiforme, quicklime, and powdered scallop shell. The diagnosis of HP was confirmed immunologically by the detection of serum precipitins to powdered Hijikia fusiforme, and by the positive result of in vitro lymphocytic proliferative response for Konjak flour using peripheral blood lymphocytes. To our knowledge, this is the first case report of Konnyaku manufacturer's lung.

Summary Background We have recently reported that soluble ST2 protein levels are elevated in the sera of patients with asthma, and correlate well with the severity of asthma exacerbation. However, the role, function, and kinetics of soluble ST2 expression in asthma remain unclear. Objective The objective of the present study was to clarify the function and kinetics of soluble murine (m) ST2 expression in a murine asthma model. Methods We analyzed the kinetics of gene and protein expression of mST2 in sera or lung tissue after allergen (ovalbumin; OVA) challenge in a murine model of allergic airway inflammation, the effects of mST2 protein on OVA-induced Th2 cytokine production in vitro from splenocytes of sensitized mice, and the effects of soluble mST2 on Th2-dependent allergic airway inflammation by in vivo gene transfer of mST2. Results Serum mST2 protein levels increased to the maximal level 3 h after the allergen challenge, before serum IL-5 levels peaked. The mRNA expression of mST2 in lung tissue was induced after the allergen challenge, while that in the spleen was constitutively detected. Furthermore, pre-treatment with mST2 protein significantly inhibited the production of IL-4 and IL-5, but not IFN-γ, from OVA-stimulated splenocytes in vitro, and intravenous mST2 gene transfer resulted in a drastic reduction in the number of eosinophils and in the levels of IL-4 and IL-5 in bronchoalveolar lavage fluid, compared with those in response to transfer of non-coding plasmid vector or of lipid alone. Conclusion These results suggest that increases in endogenous mST2 protein after allergen exposure may modulate Th2-mediated airway inflammation, and that in vivo gene transfer of mST2 can be applicable to use in a novel immunotherapy for allergic diseases.

Previous studies have reported that ST2 is preferentially expressed on Th2 cells and plays a critical part in controlling airway inflammation in murine models of asthma. However, the clinical role of ST2 in patients with bronchial asthma remains unclear. In our study, we examined 56 patients with atopic asthma in a nonattack phase and 200 nonatopic normal volunteers for healthy control, and analyzed the relationship of their serum ST2 levels to asthma severity, pulmonary function, and laboratory data. Of the 56 patients with atopic asthma, 30 exhibited asthmatic exacerbation, and their serum ST2 levels were also analyzed. The serum ST2 levels were low, but a statistical difference was found between patients with nonattack asthma and the healthy control group (p0.05). We also found a differential rise of serum ST2 level that correlates well with the severity of asthma exacerbation. Furthermore, the serum ST2 levels during asthma exacerbation statistically correlated with the percentage of predicted peak expiratory flow (r = -0.634, p = 0.004) and Pa(CO(2)) (r = 0.516, p = 0.003). These results suggest that soluble human ST2 protein in sera may be related to Th2-mediated allergic inflammation inducing acute exacerbation in patients with atopic asthma.

The human ST2 gene can be specifically induced by growth stimulation in fibroblastic cells, and can also be induced by antigen stimulation in Th2 cells. The gene encodes a soluble secreted protein, ST2, and a transmembrane protein, ST2L, which are closely related to the interleukin-1 receptor. To gain insight into the biological roles of the ST2 gene, three monoclonal antibodies (MAbs) against human ST2 gene products were obtained. To obtain these antibodies, immunization was carried out using two different immunogens: purified soluble human ST2 protein (hST2), and COS7 cells, which express the extracellular portion of human ST2L. 2A5 and FB9 MAbs were derived from the immunization with soluble hST2, and HB12 was derived from the COS7 cell immunization. All three antibodies were shown to detect native forms of the human ST2 gene products by immunoprecipitation, flow cytometry, and enzyme-linked immunosorbent assay (ELISA). In the competitive ELISA using biotinylated and nonlabelled MAbs, neither FB9 nor HB12 affected the binding of 2A5 to ST2 gene products. Based on this result, we constructed a sandwich ELISA system using 2A5 and FB9 to measure the concentration of soluble hST2 in sera. The ELISA, combined with the flow cytometry using these antibodies, will be a useful tool for elucidating the functions of human ST2 gene products in individuals.

We report here that a cancer gene therapy protocol using a combination of IL-12, pro-IL-18, and IL-1β converting enzyme (ICE) cDNA expression vectors simultaneously delivered via gene gun can significantly augment antitumor effects, evidently by generating increased levels of bioactive IL-18 and consequently IFN-γ. First, we compared the levels of IFN-γ secreted by mouse splenocytes stimulated with tumor cells transfected with various test genes, including IL-12 alone; pro-IL-18 alone; pro-IL-18 and ICE; IL-12 and pro-IL-18; and IL-12, pro-IL-18, and ICE. Among these treatments, the combination of IL-12, pro-IL-18, and ICE cDNA resulted in the highest level of IFN-γ production from splenocytesin vitro, and similar results were obtained when these same treatments were delivered to the skin of a mouse by gene gun and IFN-γ levels were measured at the skin transfection sitein vivo. Furthermore, the triple gene combinatorial gene therapy protocol was the most effective among all tested groups at suppressing the growth of TS/A (murine mammary adenocarcinoma) tumors previously implanted intradermally at the skin site receiving DNA transfer by gene gun on days 6, 8, 10, and 12 after tumor implantation. Fifty percent of mice treated with the combined three-gene protocol underwent complete tumor regression.In vivodepletion experiments showed that this antitumor effect was CD8+T cell-mediated and partially IFN-γ-dependent. These results suggest that a combinatorial gene therapy protocol using a mixture of IL-12, pro-IL-18, and ICE cDNAs can confer potent antitumor activities against established TS/A tumors via cytotoxic CD8+T cells and IFN-γ-dependent pathways.

To clarify the bronchoscopic findings in metastatic spread to the bronchi, we analyzed the records of 65 cases of metastatic pulmonary disease in which fiberoptic bronchoscopy had been done. Forty-five patients (69.2%) had abnormal bronchoscopic findings. These patients could be divided into three groups, according to bronchoscopic findings and route of metastatic spread to the bronchi: endobronchial metastasis (n=15), bronchial involvement (that is, direct extension to the bronchi from adjacent metastatic foci, n=15), and lymphangitis carcinomatosa (n=15). Breast cancer and colon cancer were common in cases of endobronchial metastasis, and the bronchial tumor often presented as a polypoid or nodular lesion covered with necrotic material. Submucosal swelling with an irregular margin and narrowing of the bronchial lumen were seen in cases of bronchial involvement. In conclusion, each type of primary extrapulmonary tumor is associated with characteristic endobronchial findings of pulmonary metastases such as endobronchial metastasis and bronchial involvement, which should be discriminated if possible, because of their different metastatic process.

The authors have investigated gene expression of ST2 in the lung tissue of a bleomycin (BLM)-induced lung fibrosis model in vivo and in a human lung fibroblast cell line, WI38, and a human type II alveolar epithelial cell line, A549, reacting to proinflammatory and type 2 helper T cell (Th2)-type cytokine stimuli in vitro. The lung mRNA expression of interleukin (IL)-4, IL-5, IL-1beta, and tumor necrosis factor (TNF)-alpha increased significantly at day 7 after instillation of BLM, whereas interferon (IFN)-gamma mRNA expression did not increase. ST2 and transforming growth factor (TGF)-beta1 mRNA expression of the lung increased significantly between days 7 and 21, and increased to maximal levels at day 14 post-BLM challenge. ST2 mRNA expression statistically correlated with TGF-beta 1 mRNA expression. In addition, the combination of IL-1 beta, TNF-alpha, and IL-4 had an additive effect on ST2 mRNA expression from A549 cells and WI38 cells. These findings suggest that soluble ST2 gene may increase, possibly reflecting the development of the inflammatory process and the Th2-type immune response in the fibrotic lung tissue, and may modulate a process of pulmonary fibrosis.

We report a rare case of multiple pulmonary metastases with cavitation from gallbladder cancer. A 77-year-old woman was admitted to our hospital complaining of productive cough and exertional dyspnea. Chest X-ray film showed multiple nodular shadows with some cavitation. Computed tomography showed multiple cavities, up to 2 cm in diameter, as well as nodules, in bilateral lung fields. Under a survey of primary focus, the ultrasonographic test of the abdomen revealed a hypoechoic mass in the hepatic hilum. The patient died of respiratory failure. Autopsy findings revealed that that multiple lung tumors had metastasized from papillary adenocarcinoma of the gallbladder and that cavitation of the lung was formed by bronchioloectasis.

Object The serum levels of KL-6, surfactant protein A (SP-A), and SP-D are useful biomarkers and prognostic factors for the activity of interstitial pneumonias. The aim of this study was to determine the clinical roles of the levels of KL-6, SP-A, and SP-D in the serum and bronchoalveolar lavage fluid (BALF) of patients with acute eosinophilic pneumonia (AEP).Materials and Methods We researched 5 cases of AEP. The levels of KL-6, SP-A, and SP-D in the sera and BALF of those patients were measured by enzyme-linked immunosorbent assay.Results KL-6 levels in BALF did not differ between AEP patients and the healthy control group, while SP-A and SP-D levels in BALF were significantly higher in the AEP patients than in the healthy control group. In sera, AEP patients had significantly higher than normal levels of SP-A and SP-D, but not of KL-6. Only in sera there was a positive correlation between SP-A and SP-D, but no apparent correlations in BALF and also between KL-6 and the others. Furthermore, the BALF levels of SP-D, but not of SP-A or KL-6, statistically correlated with the concentration of albumin in BALF. After clinical improvement, the elevated levels of serum SP-A or SP-D in AEP patients decreased until normal levels were reached within 2 months.Conclusion These results suggest that the serum or BALF levels of SP-D appear to be more sensitive than those of SP-A or KL-6 at reflecting the inflammatory response in AEP lungs.

We report two cases of lung cancer in the third decade of life. One patient was a 27-year-old man complaining of persistent cough and chest pain. He was referred for examination of an abnormal shadow found on chest radiography. A diagnosis of adenocarcinoma consisting of bronchial gland cells was made by bronchial biopsy. The clinical stage was T4N2M1. Although he was treated with cisplatin-based chemotherapy and gefitinib, he died of lung cancer. The other patient was a 26-year-old Chinese woman complaining of hemosputum. Since histologic examination at thoracotomy revealed lung cancer, left lower lobectomy and lymph node dissection were performed. Histological examination of the resected specimen revealed pleomorphic carcinoma. The pathological stage was pT2N2MX. Although the absolute number of young adults with lung cancer has been increasing, the relative incidence among all cases of lung cancer has been decreasing. In some cases, young adults with lung cancer have been misdiagnosed as having bronchial asthma or pulmonary tuberculosis.

We report a case of intrathoracic desmoid tumor without familial adenomatous polyposis and demonstrate beta-catenin mutation of exon 3. A 15-year-old male presented with a desmoid tumor after having sustained an assault. In an examination for a mutation of the beta-catenin gene, an activating mutation from ACC (Thr) to GCC (Ala) at codon 41 was found. Immunohistochemical staining showed that accumulated beta-catenin protein was predominantly localized in the nuclei of desmoid cells, and cyclin D1 protein was also overexpressed. These findings might suggest that an activating mutation of the beta-catenin gene affected regulation of the cyclin D1 gene, resulting in the generation of intrathoracic sporadic desmoid tumor, which arose at the site of posttraumatic injury.

The human ST2 gene can be specifically induced by growth stimulation in fibroblastic cells, and the soluble ST2 protein (ST2) is expressed preferentially in T-helper type 2 (Th2) cells. Furthermore, ST2 is induced by proinflammatory stimuli such as tumor necrosis factor-alpha and interleukin-1 beta. It has been reported that the inflammatory response in idiopathic pulmonary fibrosis (IPF) is thought to be associated with proinflammatory cytokines and Th2 immune response.The objective of this study was to evaluate the relevance of the serum ST2 levels in the pathogenesis of IPF.Retrospective study.Inpatients in a college hospital.Forty-nine patients with IPF admitted to our hospital 64 times: 36 patients were admitted once, 11 patients were admitted twice, and 2 patients were admitted three times. The participants also included 200 healthy control volunteers.Among 64 events in 49 patients with IPF, 50 of the events occurred in a stable state, and 14 events occurred during acute exacerbation. An acute exacerbation of IPF was defined as an accelerated phase of IPF. The serum ST2 levels were measured by enzyme-linked immunosorbent assay. The serum levels of ST2 in the stable state group did not differ from those in the healthy control group, while the serum levels of ST2 in the acute exacerbation group were significantly higher than those in the stable state group or the healthy control group (p0.001, acute exacerbation group vs stable state group or healthy control group; acute exacerbation group, 2.76 +/- 0.56 ng/mL; stable state group, 0.44 +/- 0.07 ng/mL; healthy control group, 0.42 +/- 0.03 ng/mL). Furthermore, serum ST2 statistically correlated with lactate dehydrogenase (r = 0.344, p = 0.005) and C-reactive protein (r = 0.496, p0.001), and inversely correlated with PaO(2) (r = - 0.356, p = 0.018) and the percentage of predicted vital capacity (r = - 0.346, p = 0.026).These results suggest that ST2 protein may increase in the serum, reflecting severity in the inflammatory process and Th2 immune response in the IPF lung.

Macrolides have been shown to be effective in treating diffuse panbronchiolitis (DPB), although the precise modes of action remain unclear. At sites of airway inflammation, respiratory epithelium is considered an active participant in regulating neutrophil survival. We therefore examined the effect of erythromycin, clarithromycin, azithromycin, and josamycin on both neutrophil survival and on epithelial-derived factors, which influence neutrophil longevity. Media conditioned with transiently tumor necrosis factor (TNF)-alpha-stimulated A549 human airway epithelial cells prolonged neutrophil survival compared with control media. The presence of dexamethasone during neutrophil culture led to further prolongation of neutrophil survival. In contrast, none of the tested macrolides modulated neutrophil survival, suggesting a lack of direct effect of these drugs. On the other hand, pretreatment of TNF-alpha-stimulated A549 cells by erythromycin, clarithromycin, azithromycin, or dexamethasone, but not josamycin, decreased the neutrophil survival-enhancing effects in a dose-dependent manner. Neutralizing antibodies to granulocyte macrophage colony-stimulating factor (GM-CSF) dampened the prolonged neutrophil survival observed in TNF-alpha-stimulated A549 conditioned media. Erythromycin, clarithromycin, azithromycin, and dexamethasone inhibited TNF-alpha-induced GM-CSF expression in A549 cells at both the protein and messenger RNA levels. These results suggest that macrolides inhibit epithelial cell-mediated neutrophil survival by modulating GM-CSF release, which may, at least in part, explain the effectiveness of this family of drugs on DPB.

Most cases of thoracic actinomycosis (TA) have been surgically diagnosed and treated, since it is difficult to distinguish this condition from lung cancer. To clarify the features that are useful for the radiological diagnosis of TA, we analyzed the correlation between the radiological and pathological findings. TA was histologically diagnosed in the 11 patients of the present study, who were admitted to our hospital between 1976 and 2001. In the chest CT, consolidation was observed in 5 cases, a mass-like shadow in 4, a central low attenuation area (LAA) in 7, and bronchial and bronchiolar dilatation in 6. In the pathological findings, all cases showed bronchial dilatation and abscess formation in the bronchi, as well as peribronchial granulation. The evaluation of radiological and pathological correlation revealed that the central LAA in the chest CT corresponded to the abscess formation composing of actinomyces, and the cavity corresponded to the bronchial dilatation. These results suggest that radiological findings, such as LAA and bronchial and bronchiolar dilatation, are characteristic for TA, and can be useful in making a diagnosis of TA.

The authors investigated the regulation of toll-like receptor (TLR) TLR2 and TLR4 gene expression in alveolar macrophage (AM) in response to lipopolysaccharide (LPS) or proinflammatory cytokines in vitro. Treatment of a murine AM cell line, MH-S, with LPS, tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta significantly increased TLR2 mRNA expression, whereas TLR4 mRNA expression remained constant. LPS-mediated TLR2 mRNA up-regulation was attenuated by inhibition of p38 kinase (with SB203580) or nuclear factor (NF)-kappaB (with sulfasalazine or SN-50), but not by inhibition of extracellular signal-regulated kinase (with PD98059) or c-Jun N-terminal kinase (with SP600125), suggesting that LPS may induce TLR2 mRNA expression through p38 kinase and NF-kappaB activation. These results indicate that TLR2 expression up-regulated in AM in response to bacterial respiratory infections may render AM responsive to TLR2 ligands, which may accelerate the innate immunity against pathogens in the lung.

Effective eradication of established tumors and generation of a lasting systemic immune response is an important goal for cancer gene immunotherapy. The method of gene delivery may also be critical for the generation of an effective antitumor response. We compared the level of transgene expression and antitumor activity of two different interleukin (IL)-12 DNA preparations (naked DNA versus DNA lipid complex). Established murine adenocarcinoma (CT26) and renal cell carcinoma (Renca) tumors in BALB/c mice were treated by direct intratumoral injection of a nonviral plasmid DNA vector encoding the murine IL-12 (mIL-12) gene, either alone (naked) or in complexes with cationic lipid. Both treatments resulted in the same percentage (87%) of mice undergoing a complete tumor regression of the CT26 tumor. For the Renca tumor model, complete tumor regression was observed in 67 and 75% of animals treated with naked mIL-12 DNA and mIL-12 DNA plus lipid, respectively. Mice that were rendered tumor free for50 days by mIL-12 gene therapy rejected a subsequent challenge of parental tumor cells but not of an unrelated, syngeneic tumor. The marked reduction of tumor growth in tumor-bearing mice treated with mIL-12 cDNA was associated with the augmentation of tumor-specific cytotoxic T cells, enhanced production of IFN-gamma in spleen and lymph node cells, and increased splenomegaly and lymphadenopathy. The CD8+: CD4+ ratio in tumor-infiltrating lymphocytes was significantly increased in the tumor-bearing mice treated with mIL-12 DNA alone and mIL-12 cDNA plus lipid as compared with a control vector-treated group. These results indicate that direct intratumoral gene transfer with naked nonviral IL-12 DNA provides an effective and simple method for the treatment of murine tumors, suggesting an approach for clinical application.

We have investigated gene and protein expression of ST2/ST2L in a murine alveolar macrophage (AM) cell line, MH-S, reacting to inflammatory stimuli in vitro and in the lung tissue of an acute lung injury model in vivo. We have also analyzed the effect of soluble ST2 protein on inflammatory response of MH-S cells. Lipopolysaccharide (LPS) and proinflammatory cytokines such as IL-1beta, IL-6, and TNF-alpha induced ST2 mRNA expression in MH-S cells. In an acute lung injury model, protein and mRNA expression levels of ST2 increased to the maximal level at 24-72h after the LPS challenge. Furthermore, pretreatment with ST2 protein significantly reduced the protein production and gene expression of IL-1alpha, IL-6, and TNF-alpha in LPS-stimulated MH-S cells in vitro. These results suggest that increases in endogenous ST2 protein in AM, which is induced by inflammatory stimuli, such as LPS and proinflammatory cytokines, may modulate acute lung inflammation.

The patient was a 56-year-old man who had received diagnoses of psoriasis vulgaris at the age of thirty-three and of nephrotic syndrome at forty-five, and had been treated with prednisolone. He consulted the outpatient clinic because of a cough, and interstitial pneumonia was diagnosed. He was admitted to our hospital because his symptoms and chest radiographic findings continued to worsen under treatment with antitussives. Chest radiography revealed a linear-reticular shadow in the lower lung fields. Chest computed tomography also revealed micro-cystic lesions, reticular shadows and traction bronchiectasis underneath the pleura at the back of both lower lobes. His serum titer of antinuclear antibody was increased, but he had no other symptoms or autoantibodies. Surgical lung biopsy under thoracoscopy revealed usual interstitial pneumonia (UIP). The association of interstitial pneumonia with psoriasis vulgaris is rare.

In sarcoidosis, unknown antigen(s) causes Th1-mediated granulomatous inflammation with cytokines such as IFN gamma and IL-12, initially. IL-16, IL-8, IP-10 and RANTES are participated in the accumulation of CD4+ T cell. For the chemotaxis of macrophages and monocytes, MCP-1, MIP1-alpha and RANTES are participated. Local proliferation of T cell is induced by IL-2 and IL-15 and that of macrophage/monocyte lineage is done by M-CSF, GM-CSF and G-CSF. Removal of the causative antigen(s) allows immune-suppressive cytokines such as TGF beta to downregulate the immune response and granuloma formation. Failure of removal of causative antigen(s) can induce prolonged existence of granuloma and irreversible fibrosis.

We describe a 15-year-old asymptomatic girl with multiple pulmonary nodules which turned out to be cavernous hemangiomas. The radiological findings of other organs revealed no abnormalities, excluding the liver, where multiple low-density areas were located. Thoracoscopy and laparoscopy revealed cavernous hemangiomas of the lung and liver. While liver cavernous hemangiomas are common, pulmonary hemangiomas are fairly rare. Although cavernous hemangiomas are classified as malformations, some of these lesions have been reported to grow. In pulmonary hemangiomas, some cases have an inexorable clinical course with hemoptysis, respiratory distress, and heart failure. In our case, the lesions have not remarkably progressed under observation for more than 2 years.

The objective of this study was to test the hypothesis that accumulated helper T lymphocytes in malignant pleural effusions may shift to T-helper type 2 (Th2) and produce soluble ST2 protein. We took samples of serum and pleural effusions (p-) from patients with carcinomatous pleurisy (CA, n = 17), tuberculous pleurisy (TB, n = 8), and congestive heart failure (HF, n = 5) and compared the concentration of cytokines or ST2. Ex vivo production of interleukin (IL)-4 and IL-10, though not that of interferon (IFN)-gamma or IL-12, from CD4+ T cells isolated from pleural effusions was higher in the CA group than in the TB or HF group. The p-ST2 concentrations were significantly higher in the CA group than in the TB or HF group, positively correlated with the percentage of pleural effusion CD4+ T cells (r = 0.432, p = 0.016) and inversely correlated with p-IFN-gamma concentrations (r = -0.423, p = 0.019). Furthermore, mRNA expression of ST2 in CD4+ T cells isolated from group CA was upregulated, compared with that in those isolated from the TB group. These results suggest that CD4+ T cells in CA shift to Th2, which can produce soluble ST2 protein, resulting in increased concentrations of p-ST2 in malignant pleural effusion.

lymphocytes andtheir released cytokines can contribute to eosinophilicaccumulation in the lung (1–4). ST2 is an orphan receptorexpressed preferentially on the surface of murine helper Tcell type 2 (Th2) (5,6), and has been demonstrated to playan essential role in the development of Th2 responses in amurine model of eosinophilic airway inflammation (7,8).Here we describe a case of AEP with increased levels ofsoluble ST2 protein and Th2 cytokines such as interleukin(IL)-4, IL-5, IL-6 and IL-10 in bronchoalveolar lavage fluid(BALF), all of which then decreased with corticosteroidtreatment in parallel with disappearance of BALF eosino-philia. Such results suggest that ST2 expression may beinvolved in Th2-mediated eosinophilic inflammation in thepathogenesis of AEP.

We have reported that particle-mediated interleukin 12 (IL-12) gene transfer into the skin overlying the local tumor inhibits systemic metastases. To further characterize this effect, we compared the antitumor and antimetastatic effects of IL-12 cDNA delivered at the local tumor site versus at a site distant from the primary tumor, in a spontaneous metastasis model of LLC-F5 tumor. Local IL-12 gene delivery into the skin overlying the intradermal tumor (local IL-12 treatment) on days 7, 9, and 11 after tumor implantation resulted in the most suppression of the growth of the primary LLC-F5 tumor, whereas IL-12 gene transfer into the skin distant from the tumor (distant IL-12 treatment) was less effective. In contrast, both local IL-12 and distant IL-12 treatment, followed by tumor excision, inhibited lung metastases to a similar extent, resulting in significantly extended survival of test mice. The results of in vivo studies using depleting anti-asialo GM1 antibody and anti-CD4/anti-CD8 monoclonal antibodies, or neutralizing anti-interferon gamma (IFN-gamma) monoclonal antibody demonstrated that natural killer (NK) cells, CD8(+) T cells, and IFN-gamma contributed to the antimetastatic effects in both treatment groups. Furthermore, the levels of mRNA expression of vascular endothelial growth factor and matrix methalloproteinase 9 at the tumor microenvironment were suppressed after both local and distant IL-12 treatment. These results suggest that the current particle-mediated IL-12 gene delivery in the spontaneous LLC-F5 metastasis model can confer antimetastatic activities, irrespective of the gene transfection site, via a combination of several mechanisms involving CD8(+) T cells, NK cells, IFN-gamma, and antiangiogenesis.

Pier Giuseppe Agostoni, Milano Marina Aiello, Parma Nicolino Ambrosino, Gussago Eleftherios Anevlavis, Nea Ionia Veena B. Antony, Indianapolis, Ind. Lorenzo Appendini, Veruno Hormoz Ashtyani, Hackensack, N.J. I. Azziz, Edmunds Philip Bardin, Clayton, Melbourne Harry Bassaris, Patra Eric Bateman, Groote Schuur, Cape Town Robert P. Baughman, Cincinnati, Ohio Egmont Baumgartner, Hall, Tirol F. Xaver Baur, Hamburg Robert Berent, Wels Sten Erik Bergstrom, Huddinge Andrea Bianco, Napoli Sebastiano Bianco, Milano Roland Bittner, Berlin Francesco Blasi, Milano Annette Boehler, Zurich Thomas Boehm, Zurich Josef R.M. Bolitschek, Linz T. Bombeli, Zurich Louis-Philippe Boulet, Sainte-Foy Henri Bounameaux, Geneve Demosthenes E. Bouros, Athens Homer A. Boushey, San Francisco, Calif. Lennart Braback, Sundsvall Thomas Brack, Zurich Aidan Bradford, Dublin Christian Brambilla, Grenoble Antonio Braschi, Pavia Otto M. Braun, Horn O. Brandli, Faltigberg Vita Brusasco, Genova Martin H. Brutsche, Basel Jean-Luc Burgaud, Sophia Antipolis Andrew Bush, London Cecilia Calabrese, Napoli Philippe Camus, Dijon Pier Aldo Canessa, Sarzana, La Spezia Laura Carrozzi, Pisa Antonio Castagnaro, Mantova Sergio Cavaliere, Brescia M. Cazzola, Napoli I. Cerveri, Pavia Pascal Chanez, Montpellier Neil S. Cherniack, Newark, N.J. Alfredo Chetta, Parma Gianfranco Chiari, Parma Luke Clancy, Dublin Enrico Clini, Gaiato dei Pavullo Henri Colt, La Jolla, Calif. J.H. Coote, Birmingham Giuseppe M. Corbo, Roma Antonio Corrado, Firenze Paolo Coruzzi, Parma U. Costabel, Essen J. Costello, London Nunzio Crimi, Catania Isidor Dab, Bruxelles Roberto Dal Negro, Bussolengo Filiberto Dalmasso, Torino Peter Dalquen, Basel Peter Davies, Liverpool Fernando De Benedetto, Chieti Francesco de Blasio, Arco Felice Marc Decramer, Leuven P.N. Richard Dekhuijzen, Nijmegen Jose Pablo Diaz Jimenez, Barcelona Robert Dinwiddie, London Peter Dorow, Berlin Keith Dorrington, Oxford Roberto Duranti, Firenze Michael Dusmet, London J. Eckmayr, Wels B. Emmerich, Munchen Armin Ernst, Boston, Mass. Santiago Ewig, Bonn H. Fabel, Hannover A. Fairfax, Stafford Franco Falcone, Bologna Rudolf Ferlinz, Bad Gastein Jean-William Fitting, Lausanne Pierre Fournel, St. Etienne Lutz Freitag, Hemer Dimitris Georgopoulos, Heraklion Vassilis Georgoulias, Heraklion Helen Giamarelou, Marousi, Athens Simon Godfrey, Jerusalem A. Greening, Edinburgh Paul Haber, Wien Dean Handley, Marlborough, Mass. Patricia L. Haslam, London W. Heindl, Wien Arthur Helbling, Bern Bill Hesselmar, West Perth Richard Hodder, Ottawa J.C. Hogg, Vancouver Rudolf Maria Huber, Munchen Gerard Huchon, Paris David W. Hudgel, Detroit, Mich. Stasia Jastrzembski, Newark, N.J. Amal Jubran, Maywood, Ill. Frank Kanniess, Grosshansdorf Jack Kastelik, Cottingham, East Yorkshire Philipp Kaufmann, Zurich Roland Keller, Aarau Frank Kelly, London Paul Kemp, Leeds Ok Hwa Kim, Kyonggi-do Carl M. Kirsch, San Jose, Calif. Nikolaus Konietzko, Essen D. Kohler, Schmallenberg-Grafschaft Richard Kraemer, Bern Claus Kroegel, Jena Friedrich Kummer, Wien Boris Lams, London Irene Lang, Wien Didier Lardinois, Zurich R. Lazor, Geneve Ph. Leuenberger, Lausanne Brian Lipworth, Dundee H. Lode, Berlin Stelios Loukides, Athens Maurizio Luisetti, Pavia Piero Maestrelli, Padova H. Magnussen, Grosshansdorf Katerina Malagari, Papagou, Athens Carlo Manca, Parma Milan Marel, Praha Charles-Hugo Marquette, Lille C. Marriott, London Praveen N. Mathur, Indianapolis, Ind. Heinrich Matthys, Freiburg/Br. Harald Mauch, Berlin Thomas Meisl, Wien Giovanni Migliori, Tradale M. Miniati, Pisa Anna Maria Moretti, Bari A.H. Morice, Cottingham Gianna Moscato, Pavia Michael Rolf Mueller, Wien