Increased serum high mobility group box-1 level in Churg-Strauss syndrome.

Churg-Strauss syndrome (CSS) is a rare form of systemic vasculitis occurring in patients with asthma and hypereosinophilia; however, its mechanisms involved in the severe tissue inflammation with vasculitis are poorly understood. High mobility group box 1 (HMGB1) protein, originally identified as a DNA binding protein, also has potent pro-inflammatory and proangiogenic properties. In this study, we hypothesized that HMGB1 might be associated with CSS, and examined serum HMGB1 levels and compared those of asthma patients and healthy volunteers. We also investigated HMGB1 expression in the lesion, and eosinophil HMGB1 amount in CSS patients. We found that the serum HMGB1 levels in CSS patients were significantly higher than those of asthma patients and healthy volunteers. Eosinophils in the CSS lesion expressed HMGB1 and HMGB1 level in eosinophils from CSS patients was significantly higher than that of asthma patients, while there was no significant difference in HMGB1 levels in peripheral mononuclear cells. The serum HMGB1 level in CSS patients decreased after the steroid therapy, and showed significant positive correlations with several molecules, including soluble interleukin-2 receptor, soluble thrombomodulin, and eosinophil cationic protein in sera. We propose that HMGB1 might contribute to the pathogenesis of CSS.

Clinical investigation: increased serum stromal derived factor 1 alpha levels in pulmonary tuberculosis.

Pulmonary tuberculosis, a granulomatous disease, has few serological markers for its activity. Recently, an increased plasma level of stromal derived factor 1 alpha (SDF-1alpha), which can induce strong chemotaxis of cells through its receptor CXCR4, was detected in patients with tuberculosis. In this study we investigated serum SDF-1alpha levels and CXCR4 expression on peripheral blood mononuclear cells (PBMCs). Fifty-five active tuberculosis patients, 30 resolved tuberculosis patients, 27 acute bronchitis patients and 8 healthy volunteers were examined. Histological expression of SDF-1alpha in the tuberculosis lesion and CXCR4 expression of PBMCs were also analysed. Serum SDF-1alpha levels in active tuberculosis patients were significantly higher than other groups. The sensitivity and specificity for the diagnosis of active tuberculosis was 88.5% and 85.3% (cutoff value = 650 pg/ml), respectively. CXCR4 expression levels on PBMCs showed a significant negative correlation with serum SDF-1alpha levels. Inflammatory cells including multinuclear giant cells in the lesion expressed SDF-1alpha. Measurement of serum SDF-1alpha could be a useful screening marker for the identification of active pulmonary tuberuculosis. We propose that interaction of SDF-1alpha and CXCR4 might be involved in the pathogenesis of pulmonary tuberculosis.

Influence of human T lymphotrophic virus type I on diffuse pan-bronchiolitis.

Human T lymphotrophic virus type-I (HTLV-I), a human retrovirus, infects CD4(+) lymphocytes and is thought to modify their function and a possible association with pulmonary diseases has also been suggested. However, little is known about the influence of HTLV-I on diffuse pan-bronchiolitis (DPB), a chronic inflammatory lung disease with infiltration of lymphocytes and hyperplasia of the bronchus-associated lymphoid tissue. In this study, 35 DPB patients with and without HTLV-I infection were examined. HTLV-I positive DPB patients were likely to have a larger affected area with lower FEV(1). The CD3(+)/CD25(+) lymphocyte percentage was significantly higher in the BALF of HTLV-I positive patients than in negative patients. MIP-1 alpha, IP-10 and levels in BALF were also significantly higher in HTLV-I positive patients than in negative patients. The levels of MCP-1 and IL-8 were not significantly different. In HTLV-I positive patients, the MIP-1 alpha and IP-10 levels showed a significant positive correlation with the percentage of CD3(+)/CD25 lymphocytes. BALF cells of all HTLV-I positive DPB patients showed expression of p40(tax) mRNA. We suggest that HTLV-I infection may modify DPB pathogenesis via activation of T cells. We also found that the frequency of ATL development in HTLV-I positive DPB patients was significantly higher than in all HTLV-I positive patients (OR = 8.22, 95% CI = 2.61-25.9, P < 0.01). The levels of TGF-beta in patients who developed ATL were significantly lower than in patients who did not develop ATL. Sensitivity and specificity were 80% and 85.7%, respectively (cut-off = 20 pg/ml). We also propose that these features should be taken into consideration in the treatment of DPB in HTLV-I infected individuals.

Signaling through the beta2 integrin prolongs eosinophil survival.

BACKGROUND: Recently, adhesion molecules have been suggested to play an important role in allergic inflammatory diseases such as bronchial asthma. It is unclear whether eosinophil activation and paracrine or autocrine synthesis of eosinophilopoietic growth cytokines is mediated through signaling by intercellular adhesion molecule-1 (ICAM-1) and the beta2 integrin family. OBJECTIVE: We examined whether signaling by ICAM-1 and its ligands (beta2 integrins) could prolong eosinophil survival. METHODS: Eosinophils were isolated from patients with hypereosinophilia by modified CD16 negative selection. After culture with or without recombinant soluble ICAM-1, eosinophil viability was measured by trypan blue dye exclusion. RESULTS: Eosinophil survival was prolonged in cultures with recombinant soluble ICAM-1 compared with cultures without it (P <.01 on days 2, 4, and 6); this effect was dose-dependent. Eosinophil survival in cultures with recombinant soluble ICAM-1 was significantly inhibited by antibodies against ICAM-1 (P <.01), complement receptor 3 (P <.01), and lymphocyte function-associated antigen-1beta (P <.01). Anti-IL-3 showed no effect on eosinophil survival, whereas anti-IL-5 caused partial inhibition of survival. Interestingly, anti-granulocyte/macrophage colony-stimulating factor caused the complete inhibition of eosinophil survival in cultures with recombinant soluble ICAM-1. CONCLUSION: These results suggested the importance of the beta2 integrins in eosinophil-mediated allergic inflammation.

[An autopsy case of neurogenic pulmonary edema due to suicide by hanging].

A 43-year-old man died of pulmonary edema after hanging himself. His cardiac function and renal function were normal. No aspiration, trauma, or overhydration was present, and high doses of catecholamine had not been administered. Autopsy findings revealed hypoxic brain damage, cerebral edema and pyknosis of nerve cells in the medulla oblongata. These findings resulted in a diagnosis of neurogenic pulmonary edema. To our knowledge, there have been no previous reports of neurogenic pulmonary edema due to airway obstruction.

[Multiple pulmonary nodules due to ticlopidine-induced pneumonitis].

A 54-year-old man had been treated with Ticlopidine for antithrombotic therapy after a myocardial infarction. Six months after the start of Ticlopidine, pruritic skin rash developed. After 8 months, chest X-ray films revealed multiple nodules, and the patient was admitted to our hospital. Laboratory data indicated liver dysfunction and sputum eosinophilia. Transbronchial biopsy specimens disclosed intraluminal organization and alveolar septal thickening with lymphocyte infiltration. After cessation of Ticlopidine, the cutaneous lesions quickly improved, and multiple nodules completely resolved within 4 months. These findings resulted in a diagnosis of Ticlopidine-induced pneumonitis. The radiographic pattern of multiple pulmonary nodules was very rare. Sputum eosinophilia may be helpful in the diagnosis of drug-induced pneumonitis. To our knowledge, there have been no previous reports of Ticlopidine-induced pneumonitis in Japan.

Adhesion to fibronectin regulates expression of intercellular adhesion molecule-1 on eosinophilic cells.

BACKGROUND: Adhesion molecules may play an important role in eosinophilic activation as well as adherence of extracellular matrix (ECM) including fibronectin (FN) in the inflamed focus. Tissue eosinophils expressed inter-cellular adhesion molecule-1 (ICAM-1, CD54), whereas peripheral blood eosinophils did not. The molecular mechanisms of ICAM-1 expression on eosinophils are not clear. We immunologically examined the effect of adherence to FN on the expression of adhesion molecules in an eosinophilic cell line (EoL-1). METHODS: EoL-1 cells, suspended at a concentration of 2 x 10(6) cells/ml, were cultured at 37 degrees C in BSA or FN-coated 24-well plates. After 4 h, the cells were removed by pipetting, and the expression of adhesion molecules was evaluated by immunofluorescence. RESULTS: Adherence of EoL-1 to FN enhanced ICAM-1 (CD54) expression on EoL-1 (FN vs. BSA: 84.76 +/- 17.90 vs. 43.73 +/- 7.60, mean fluorescent intensity). Regarding other adhesion molecules on EoL-1 (CD11a, CD18, CD49d), the expression was not significantly augmented by adherence to FN. CONCLUSIONS: We concluded that the signal via adhesion of FN regulates the expression of ICAM-1 on eosinophilic cells. This study suggests that eosinophilic adhesion to ECM via adhesion molecules plays an important role in the pathogenesis of allergic inflammation through eosinophilic functional changes, including regulation of expressive adhesion molecules such as ICAM-1.

Eosinophilic pneumonia with eosinophilic gastroenteritis.

A 48-year-old man was admitted to our hospital with cough, fever and dysphagia. He had a past history of bronchial asthma and surgery for nasal polyp. Chest radiograph and computed tomography showed atelectasis in the right lower field and infiltrative shadow in the left lower field and overall thickening of the esophageal wall. Transbronchial lung biopsy (TBLB) specimens revealed infiltration of eosinophils and lymphocytes under the bronchial mucosa. Gastrointestinal tract biopsy specimens showed submucosal infiltration of eosinophils. These findings led to a definite diagnosis of eosinophilic pneumonia associated with eosinophilic gastroenteritis, a disease which has been rarely reported.

RANTES augments eosinophil lucigenin-dependent chemiluminescence.

BACKGROUND: RANTES (regulated on activation, normal T expressed and secreted) has been shown to possess chemotactic activity for eosinophils. Eosinophils have been considered to play a key role in the allergic inflammation through the release of inflammatory molecules such as radical oxygen products. Thus, in this study, we examined the effect of RANTES on radical oxygen products from eosinophils. METHODS: Eosinophils were isolated from heparinized venous blood of patients with bronchial asthma by the modified CD16-negative depletion method. Radical oxygen products were examined in terms of lucigenin-dependent chemiluminescence. To a mixture of 50 microl of eosinophils (2x10(6)/ml) and 50 microl of lucigenin (5x10(-4)M), 50 microl of calcium ionophore A23187 (final concentration 10(-5)M) was added, and radical oxygen products were determined for 600 s. RESULTS: RANTES treatment resulted in the enhancement of peak value (0.64+/-0.23 RLU) and integrated value (119.08+/-20.52 RLU) as compared to untreated cells (0.15+/-0.03 RLU, 29.48+/-8.92 RLU, respectively). CONCLUSIONS We could conclude that RANTES might play an important role in the pathogenesis of allergic inflammation through involvement in selective eosinophil infiltration and eosinophil activation by augmentation of eosinophil oxidative metabolism.

[A case of tuberculous abscess in the chest wall close to the thickening pleural lesion following tuberculous pleuritis].

A 33-year-old woman with a history of right tuberculous pleuritis was successfully treated in December 1992 by administration of anti-tuberculous drugs, she demonstrated residual localized pleural thickening on chest computed tomography (CT) and gradually developed a subcutaneous mass in the right chest which became apparent in March 1993. In September, chest CT revealed a periocostal abscess in the right anterior chest wall close to the localized pleural thickening. The patient was diagnosed with tuberculous abscess in the right chest wall on confirmation of acid-fast bacilli in a needle aspiration material of the abscess, and was referred to our hospital. Anti-tuberculous chemotherapy was continued but the chest abscess grew, so on January 28, 1994 she underwent a resection of the abscess, the third costal cartilage and bone, and the parietal pleural lesion connected to the abscess. Histopathological examination showed that the abscess and parietal pleural lesion were compatible with tuberculosis, i.e. both lesions consisted of caseous necrosis and epitheloid cell granuloma, but acid-fast bacilli were not demonstrated in both lesions. After one year of postoperative anti-tuberculous chemotherapy, she was followed without any therapy for 3 years and there has been no recurrence to date. When a localized thickening pleural lesion remains after tuberculous pleuritis, complication of tuberculous abscess in the chest wall should be considered.

[Clinical features of allergic granulomatosis and angiitis (Churg-Strauss syndrome). Association with asthma symptoms].

We reported clinical and laboratory findings of 5 patients with Churg-Strauss syndrome (CSS), especially association with asthma symptoms. Subjects included 3 males and 2 females with a mean age of 53.8 year-old. In all 5 patients symptoms of neuropathy; mononeuritis multiplex and in some patients, other vasculitic symptoms; fever, diarrhea, abdominal pain and skin eruptions, were noted. These clinical features and laboratory findings; marked peripheral eosinophilia and elevation of serum ECP were normalized after steroid therapy. We investigated the relation between the occurrence of CSS and the symptoms of asthma. The mean duration of asthma in this series was 17.2 years, and 4 cases were atopic and one was non-atopic asthma. In previous publications, asthmatic symptoms were severe at the onset of the disease and progressed thereafter. In our 5 cases, however, the severities of bronchial asthma were mild of two cases, moderate of two and severe of only one, moreover severe asthmatic attacks were shown in only 2 cases when the manifestation of systemic vasculitis occurred. In conclusion, although CSS has been thought that one of complications of bronchial asthma, the occurrence of CSS are not necessarily correlated with symptoms of bronchial asthma.

Eosinophil cationic protein induces insulin-like growth factor I receptor expression on bronchial epithelial cells.

Although the biologic activity of eosinophil-specific granule proteins, e.g. eosinophil cationic protein (ECP) has been attributed to cytotoxic properties, it has also been shown to be attributable to non-cytotoxic activation of various inflammatory cells. In addition, airway epithelial cells have been reported to express insulin-like growth factor I (IGF-I) receptor and proliferate in response to IGF-I. The present study examined the regulation of expression of IGF-I receptor on bronchial epithelial cells by eosinophil granule proteins was examined. ECP significantly induced IGF-I receptor expression on bronchial epithelial cells compared with unstimulated cells. These findings suggest that eosinophils participate in the repair process of injured bronchial epithelial cells in the inflamed focus through augmentation of IGF-I receptor expression by suboptimal degranulation of eosinophil granule proteins, e.g. ECP.

Regulation of eosinophil cell death by adhesion to fibronectin.

Fibronectin is an extracellular matrix (ECM) that binds to very late antigen-4 (a beta 1 integrin molecule) expressed by eosinophils. To investigate the effect of adherence to fibronectin on regulation of eosinophil cell death, survival of eosinophils was examined by trypan blue exclusion and Fas antigen expression on the cell surface using an eosinophilic cell line (EoL-1). Adhesion to fibronectin resulted in prolongation of eosinophil survival (fibronectin vs. bovine serum albumin (BSA), 62.9 +/- 5.10 vs. 53.9 +/- 4.30% viability at 72 h) and the decreasement of Fas antigen expression on EoL-1 (fibronectin vs. BSA, 24.3 +/- 2.15 vs. 74.5 +/- 8.25% positive). These findings suggest that eosinophil adhesion to ECM via adhesion molecules plays an important role in the pathogenesis of allergic inflammation, which involves eosinophil accumulation at the inflammatory site, through regulation of eosinophil cell death.

[Adhesion to fibronectin regulates expression of adhesion molecules on eosinophils].

To elucidate the mechanism for the accumulation of eosinophils through adherence in allergic inflammation, we investigate whether adhesion of eosinophils to extracellular matrix (ECM) regulates the expression of adhesion molecules on eosinophils. Highly purified eosinophils obtained from asthmatic patients were cultured on BSA or fibronectin (FN) coated plates for 4 or 48 hours. After then, removed cells were examined about the expression of each adhesion molecule. Cultured on FN resulted in the increment of the expressive CR 3 (CD11b) (BSA vs FN; MFI ratio 10.50 vs 12.09 p < 0.05), and decrement of the expressive VLA-4 (CD49d) on the eosinophils (3.93 vs 2.91 (4 h) p < 0.05, 2.86 vs 2.09 (48 h) p < 0.05). Thus, we could conclude that eosinophils adhesion to ECM via adhesion molecules might play an important role in the pathogenesis of allergic inflammation through the involvement in the regulation of the expression of adhesion molecules on eosinophils.

Effect of RANTES on intracellular expression of EG2 antigen in eosinophils.

RANTES, which is released from thrombin-stimulated platelets, is a member of the 8-kDa cytokine family that has been shown to possess chemotactic activity for eosinophils. The effect of RANTES on the intracellular expression of EG2 antigen in eosinophils was examined in this study. RANTES augmented the intracellular expression of EG2 antigen in eosinophils. These findings suggest that RANTES not only modulates the chemotactic activity of eosinophils but also intensifies the function of eosinophil activation.

Adhesion to fibronectin augments eosinophil radical oxygen products.

Fibronectin, an extracellular matrix component, is a ligand for very late activation antigen (VLA)-4, which is one of the beta 1-integrin family of molecules expressed by eosinophils. This study examined the effect of adherence to fibronectin on radical oxygen products from eosinophils. Adhesion of eosinophils to fibronectin resulted in enhancement of eosinophil production of radical oxygen species, as determined by luminol-dependent chemiluminescence of eosinophils stimulated with calcium ionophore. It was concluded that eosinophil adhesion to extracellular matrix via adhesion molecules may be important in the pathogenesis of allergic inflammation through eosinophil activation.

Increased eosinophil oxidative metabolism by treatment with soluble intercellular adhesion molecule-1.

Adhesion molecules may play an important role not only in adherence of inflammatory cells (particularly eosinophils) to an inflamed focus but also in activation of these cells. It is therefore of interest to evaluate eosinophil activation via intercellular adhesion molecule-1 (ICAM-1) and the beta 2-integrin family, namely CR3 (Mac-1), lymphocyte function-associated antigen (LFA)-1 alpha and LFA-1 beta, which are ligands for ICAM-1. Reactive oxygen species generated by eosinophils have also been considered capable of causing airway injury at the inflamed focus. This study examined the effect of recombinant soluble ICAM-1 and its ligands on eosinophil-induced radical oxygen products in terms of luminol-dependent chemiluminescence. Recombinant soluble ICAM-1 augmented eosinophil oxidative metabolism. It was concluded that signaling via adhesion molecules might play an important role in the pathogenesis of allergic inflammation through activation of eosinophils, e.g. an increase in oxidative metabolism.

Possible release of eosinophil granule proteins in response to signaling from intercellular adhesion molecule-1 and its ligands.

The presence of a large variety of membrane receptors and the identification of cytotoxic molecules (mainly granule basic proteins) have indicated that eosinophils should br considered as effector cells. It has recently been suggested that adhesion molecules, particularly intercellular adhesion molecule-1 (ICAM-1), play an important role in allergic inflammation, for example in bronchial asthma. This study therefore investigated the possible release of granule protein in response to signaling from ICAM-1 and its ligands. The concentrations of eosinophil cationic protein and eosinophil-derived neurotoxin in supernatants of eosinophils were significantly greater (p < 0.05) in the presence of recombinant soluble ICAM-1 than without it. These results suggest that signaling from ICAM-1 and its ligands might induce eosinophil activation and might be involved in degranulation of eosinophil granule proteins, e.g. eosinophil cationic protein and eosinophil-derived neurotoxin.

Induction of beta 2 integrin expression on an eosinophilic cell line (EoL-1) by the supernatant of mononuclear cells stimulated with specific allergen from asthmatic patients.

Adhesion molecules recently have been considered to play an important role in inflammatory processes in bronchial asthma. Our previous study revealed high expression of beta 2-integrin family (CR3, LFA-1 alpha, CD18) on hypodense eosinophils. Thus, from the point of view of cell-to-cell interaction between mononuclear cells and eosinophils, we examined whether the supernatant of mononuclear cells obtained from mite-allergic asthmatic patients cultured with specific allergen mite-allergen is involved in adhesion molecule expression using an eosinophilic cell line (EoL-1). These characteristics of beta 2-integrin family expression (high expression of beta 2 integrin) were induced by the supernatant of mononuclear cells from mite-allergic asthmatic patients stimulated with mite-allergen as well as with a combination of the recombinant eosinophilopoietic growth cytokines (IL-3, GM-CSF and IL-5). Thus, we could conclude that some cytokines produced by specific allergen stimulated mononuclear cells in asthmatics might be involved in allergic inflammation through the induction of adhesion molecule expression on eosinophils in asthma or allergic disorders.

[Effect of platelet-activating factor (PAF) on eosinophil adhesion to plasma-coated glass using eosinophilic cell lines (EoL-1, EoL-3)].

Adhesion molecules on various inflammatory cells, especially on eosinophils are now considered to play an important role in allergic inflammations such as bronchial asthma. In this study, we examined the effect of platelet-activating factor (PAF) on eosinophil adhesion to plasma-coated glass using eosinophilic cell lines (EoL-1, EoL-3), since PAF might be an important mediator as an eosinophil chemotactic agent in bronchial asthma. PAF markedly augmented adherence to plasma-coated plates. This finding suggests that PAF might up-regulate the inflammatory reaction in bronchial asthma through its action as an augmentative agent for eosinophil adhesion as well as its action as an eosinophil chemotactic agent.