Serum YKL-40 as predictor of outcome in hypersensitivity pneumonitis.

YKL-40, a chitinase-like protein mainly secreted by macrophages, neutrophils and epithelial cells, is increased in patients with idiopathic interstitial pneumonia and sarcoidosis. We aimed to investigate the role of YKL-40 as a biomarker in hypersensitivity pneumonitis (HP).72 HP patients, 100 interstitial lung disease (ILD) controls and 60 healthy controls were studied. YKL-40 was measured by ELISA in serum and bronchoalveolar lavage fluid (BALF) at baseline and follow-up. The relationship between YKL-40 levels, clinical variables and disease outcome was evaluated.Baseline serum YKL-40 levels were significantly higher in HP patients than in healthy controls (p<0.001), but lower than in patients with other ILDs. Baseline BALF YKL-40 levels in HP patients were the highest among ILD patients. In HP patients, serum YKL-40 correlated with the diffusing capacity of the lung for carbon monoxide at baseline (p<0.01) and over time (p<0.001). HP patients whose disease progressed or who died had higher baseline YKL-40 levels than those who remained stable and survived (p<0.001). At a cut-off of 119 ng·mL-1, the baseline serum YKL-40 level predicted disease progression (hazard ratio 6.567; p<0.001), and at a cut-off of 150 ng·mL-1 was associated with mortality (hazard ratio 9.989; p<0.001).Serum YKL-40 may be a useful prognostic biomarker in HP patients.

CCL18 in serum, BAL fluid and alveolar macrophage culture supernatant in interstitial lung diseases.

BACKGROUND: CCL18 is a CC chemokine produced mainly by antigen-presenting cells, and is chemotactic predominantly for T-lymphocytes. CCL18 can stimulate pulmonary fibroblasts and increase the collagen production in vitro. OBJECTIVES: This study aimed to compare the CCL18 levels in a variety of human biological fluids between various interstitial lung diseases (ILDs), and to reveal potential correlations with BAL cell differentials. METHODS: Serum and bronchoalveolar lavage fluid (BALF) samples were collected from 199 patients with idiopathic pulmonary fibrosis (IPF), idiopathic non-specific interstitial pneumonia (iNSIP), respiratory bronchiolitis interstitial lung disease/desquamative interstitial pneumonia (RB-ILD/DIP), cryptogenic organizing pneumonia (COP), hypersensitivity pneumonitis (HP) or sarcoidosis. Alveolar macrophage (AM) culture was performed in 44 patients with IPF, iNSIP, COP, HP, sarcoidosis or non-ILDs. The CCL18 levels in serum, BALF and AM culture supernatant were measured with ELISA. RESULTS: Both serum and BALF CCL18 levels in all ILDs were higher than in controls (all p < 0.005). In HP, CCL18 serum levels were the highest of all ILDs, and its BALF levels were significantly higher than in other ILDs except iNSIP. The BALF CCL18 levels markedly correlated with BAL cell differentials, especially with the percentage of BAL lymphocytes. In AM culture supernatant, the spontaneous CCL18 production was higher in HP and COP than in IPF and controls. CONCLUSION: CCL18 levels in serum, BALF and AM culture supernatant are markedly increased in various inflammatory and fibrotic ILDs. However, the CCL18 level being highest in HP among the investigated ILDs suggests that CCL18 may be more profoundly involved in inflammatory immune responses.

Macrolides inhibit cytokine production by alveolar macrophages in bronchiolitis obliterans organizing pneumonia.

BACKGROUND AND OBJECTIVE: Bronchiolitis obliterans organizing pneumonia (BOOP) is a distinct clinicopathological entity histologically characterized by intra-alveolar granulation tissue and absence of extensive fibrotic lesions. Effective macrolide treatment of BOOP has been reported anecdotally. This study aimed to investigate whether alveolar macrophages (AMs) produce aberrant proinflammatory cytokines in BOOP and whether this can be inhibited by clarithromycin (CAM) or azithromycin (AZM). METHODS: AMs collected by bronchoalveolar lavage (BAL) from 6 BOOP patients and 8 non-ILD controls were cultured for 24h in the presence or absence of CAM, AZM, lipopolysaccharide (LPS), or dexamethasone (DEX). Tumor necrosis factor alpha (TNF-α), soluble TNF receptor 1 (sTNFR1), sTNFR2, interleukin 1beta (IL-1ß), IL-6, IL-8, IL-10, interferon gamma inducible protein 10 (IP-10) and CC chemokine ligand 18 (CCL18) were measured in the culture supernatant by ELISA. RESULTS: The spontaneous and LPS-stimulated production of all investigated cytokines by AMs was significantly increased in BOOP compared to controls. CAM and AZM induced a dose-dependent suppression of spontaneous TNF-α, sTNFR2, IL-6, IL-8 and CCL18 production (p<0.05). CAM also inhibited the IL-1ß production. CAM and AZM significantly and dose-dependently attenuated the LPS-stimulated production of sTNFR1, sTNFR2, IL-8 and CCL18 (p<0.05). CAM also inhibited the LPS-stimulated TNF-α, IL-1ß, IL-6 and IL-10 production. CONCLUSIONS: AMs from BOOP patients produce abundant proinflammatory cytokines which may be pivotal in the disease pathogenesis. Macrolides inhibit this cytokine production, CAM more efficiently than AZM.

Increased expression of tumor necrosis factor receptors in cryptogenic organizing pneumonia.

BACKGROUND: TNF receptors (TNFR1 and TNFR2) and Fas belong to the system of apoptosis-signalling receptor molecules and may play a role in the pathogenesis of interstitial lung disease. Patients with cryptogenic organizing pneumonia (COP) usually respond well to corticosteroids, in contrast to those with idiopathic pulmonary fibrosis (IPF). This may be due to the different pathogenesis. METHODS: The expression of TNFR1, TNFR2 and Fas on bronchoalveolar lavage (BAL) macrophages and lymphocytes was analysed in 9 patients with COP, 10 with IPF and 12 controls. The production of soluble TNFR1, 2 and TNF-α by alveolar macrophages was measured by ELISA. RESULTS: TNFR1 and Fas expression on alveolar macrophages was significantly higher in COP than in controls and IPF. The expression of TNFR2 on alveolar macrophages was also increased in COP compared to controls. The expression of TNFR2 and Fas on lymphocytes was significantly higher in COP than in IPF and controls. In addition, the expression of TNFR1, TNFR2 and Fas on BAL cells correlated positively with BAL lymphocytes (p < 0.05 or p < 0.01). The production of sTNFR1 and 2 and TNF-α by macrophages in vitro was significantly increased in patients with COP compared to IPF and controls, spontaneously or with LPS stimulation (p < 0.05 or p < 0.01).There was a positive correlation between the spontaneous production of sTNFR2 and TNF-α (r = 0.494, p < 0.01). CONCLUSIONS: This study showed an increased expression of TNF receptors and Fas on BAL cells in COP that may be indicative of the local inflammatory activity in the lung. The biologic effects of this expression needs further investigation.

Angiogenic and angiostatic chemokines in idiopathic pulmonary fibrosis and granulomatous lung disease.

BACKGROUND: Angiogenesis-angiostasis balance and leukocyte recruitment are influenced by different concentrations of distinct chemokines. OBJECTIVE: To investigate the relative contribution of angiogenic and angiostatic CXC chemokines to the pathogenesis of idiopathic pulmonary fibrosis (IPF) and granulomatous lung diseases, we examined the in vitro production of an angiogenic chemokine (IL-8), and 2 angiostatic chemokines (IP-10 and MIG) by alveolar macrophages. METHODS: Alveolar macrophages from 16 patients with granulomatous lung diseases [8 with sarcoidosis, 8 with extrinsic allergic alveolitis (EAA)], 16 patients with IPF, and 8 control subjects were cultured for 24 h. IL-8, IL-18, IP-10 and MIG in the culture supernatants were measured by a fluorescent bead-based multiplex technique. RESULTS: In IPF patients, IL-8 was increased and correlated with bronchoalveolar lavage (BAL) neutrophils, whereas the levels of IP-10 and MIG were normal. In sarcoidosis and EAA patients, IL-8, IP-10, and MIG were all increased and IP-10 and MIG correlated with IL-18, a Th1 cytokine, and the percentage and number of BAL lymphocytes. CONCLUSIONS: The difference in the expression of CXC chemokines and a Th1 cytokine may contribute to the different immunopathogenesis, clinical course and responsiveness to treatment of these diseases.

Interleukin 12, interleukin 18, and tumor necrosis factor alpha release by alveolar macrophages: acute and chronic hypersensitivity pneumonitis.

BACKGROUND: Hypersensitivity pneumonitis (HP) is characterized by a granulomatous inflammation and may show various forms of clinical presentation, such as the acute, subacute, and chronic forms. The TH1-associated cytokines interleukin (IL) 12 and IL-18 and tumor necrosis factor alpha (TNF-alpha) may be involved in the pathogenesis of both the acute and chronic forms of HP. OBJECTIVE: To compare the release of IL-12, IL-18, and TNF-alpha from bronchoalveolar lavage (BAL) macrophages in these 2 forms of HP. METHODS: Patients underwent BAL 0 to 6 days after the last antigen exposure. Alveolar macrophages (AMs) from BAL in 6 patients with acute HP, 16 with chronic HP, and 11 controls were cultured for 24 hours. Cytokines in the culture supernatants were measured by enzyme-linked immunosorbent assay. RESULTS: The production of IL-12, IL-18, and TNF-alpha by AMs was increased in patients with both acute and chronic forms in either the absence or presence of lipopolysaccharide compared with controls. The levels of IL-12, IL-18, and TNF-alpha showed no difference between patients with acute and chronic HP. The spontaneous production of IL-12, IL-18, and TNF-alpha did not correlate with the CD4/CD8 ratio in BAL. The spontaneous and lipopolysaccharide-stimulated release of IL-12 showed a positive correlation with the percentage of lymphocytes (r = .470, P = .03; r = .496, P = .02; respectively) in BAL. CONCLUSIONS: This study demonstrates that an increased release of IL-12, IL-18, and TNF-alpha by AMs is associated with both the acute and chronic forms of HP.

[Transitional bladder carcinoma TaG1: role of the EGFr tyrosin kinase activation in the cellular proliferation]. / Carcinoma transicional de vejiga no invasivo y bien diferenciado: papel de la activación tirosincinasa del receptor del factor de crecimiento epidérmico en la proliferación celular.

BACKGROUND AND OBJECTIVE: The purpose of this work was to study if the expression of EGFr oncoprotein and the rate of tumoral cell proliferation in TaG1 transitional bladder carcinoma were related and if this relationship influences the potential aggressivity of the neoplasia. PATIENTS AND METHOD: Twenty-eight patients with TaG1 (non-invasive papillary and well differentiated) transitional bladder carcinoma were randomly selected. EGFr was determined by a semiquantitative immunohistochemical method in three intensity levels. The proliferative tumoral cell activity was determined by the PCNA index, that is, the relation between the total number of tumor cells and the number of tumor cells with nuclear PCNA-immunoreactivity expressed in percentage. RESULTS: The rate of cellular tumour proliferation (PCNA index) was significantly higher in those tumors with EGFr's greater expression (p=0.005). The EGFr expression was associated with tumor recurrence (p=0.012). CONCLUSIONS: The major expression of EGFr oncoprotein in tumours with higher rate of cell proliferation indicates that it may be influenced by the activity of such oncoprotein. The binding of a ligand to the EGFr leads to intracellular tumor proliferation, hence possibly favouring a more aggressive biological behavior.

Carcinoma transicional de vejiga no invasivo y bien diferenciado: papel de la activación tirosincinasadel receptor del factor de crecimiento epidérmico en la proliferación celular / Transitional bladder carcinoma TaG1: role of the EGFr tyrosin kinase activation in the cellular proliferation

FUNDAMENTO Y OBJETIVO: El objetivo de este trabajo es estudiar si la expresión de la oncoproteína transmembrana EGFr (receptor del factor de crecimiento epidérmico) y las variaciones en la tasa de proliferación celular tumoral en el carcinoma transicional de vejiga papilar no invasivo y bien diferenciado (TaG1) tienen relación entre sí e influyen, modificando en cada caso, la agresividad potencial de la neoplasia. PACIENTES Y MÉTODO: Se analizó a 28 pacientes con carcinoma transicional de vejiga TaG1 seleccionados aleatoriamente. La determinación de EGFr se realizó mediante un sistema de gradación semicuantitativo en tres niveles de intensidad de inmunotinción. La determinación de la actividad proliferativa celular tumoral se determina mediante índice de PCNA (antígeno de proliferación celular nuclear): relación entre el número total de células tumorales y el número de células con núcleos inmunorreactivos al PCNA expresado en porcentaje. RESULTADOS: La tasa de proliferación celular tumoral (índice de PCNA) fue significativamente más elevada (p = 0,005) en los tumores con mayor expresión de EGFr. La expresión de EGFr se asoció (p = 0,012) con recurrencia tumoral. CONCLUSIONES: La mayor expresión de la oncoproteína EGFr en los tumores con elevada actividad proliferativa celular indica que estaría influida por la actividad de dicha oncoproteína. La unión del ligando con el EGFr produciría la activación de la actividad tirosincinasa intracelular, que desencadenaría una cascada de señales que conducirían a la proliferación celular tumoral y favorecerían un comportamiento biológico más agresivo (AU)