Circulating levels of both Th1 and Th2 chemokines are elevated in patients with sarcoidosis

BACKGROUND: In sarcoidosis, the T helper type 1 (Th1) response tends to predominate at affected disease sites; however, whether Th1/Th2 polarization occurs in the peripheral circulation is unknown. METHODS: Fifty-two patients with sarcoidosis and 21 healthy volunteers were investigated. The concentrations of interferon-inducible protein 10 (IP-10)/CXCL10 and thymus- and activation-regulated chemokine (TARC)/CCL17 in the serum, bronchoalveolar lavage fluid (BALF) and culture supernatant were measured by an enzyme-linked immunosorbent assay. The circulating CXCR3+ CD4+ T cells and CCR4+ CD4+ T cells were assessed by flow cytometry. RESULTS: The CXCR3- or CCR4-positive ratios among CD4+ T cells were both higher in sarcoidosis than in healthy volunteers. The serum levels of both IP-10 and TARC of the patients with sarcoidosis were significantly higher than those of the healthy volunteers. In patients with sarcoidosis, a larger amount of IP-10 was generated by the BALF cells, whereas IP-10 production by peripheral blood mononuclear cells did not increase in comparison to the control subjects. The TARC levels produced by peripheral blood mononuclear cells of sarcoidosis patients were significantly higher than those of the controls, while no difference existed between the 2 groups regarding TARC production by BALF cells. CONCLUSION: IP-10 is mainly produced at the lung and TARC in the peripheral circulation in sarcoidosis patients. Both IP-10 and TARC cooperatively play a role in the pathogenesis of sarcoidosis.     

Antibody activity to Propionibacterium acnes in bronchoalveolar lavage fluid in sarcoidosis

While increased levels of circulating antibody to various microorganisms have been reported in sarcoidosis patients, the pathogenesis of the disease is still unknown. In this report, the levels of antibody activities against Propionibacterium acnes (P. acnes) were measured in bronchoalveolar lavage fluid (BALF) in patients with sarcoidosis, using an enzyme-linked immunosorbent assay method. Each immunoglobulin class of antibody activity to P. acnes was corrected by albumin concentrations in BALF. The levels of whole immunoglobulin antibody activities to P. acnes in BALF were as follows: 412.3 +/- 443.9 O.D./albumin 1 mg (M +/- SD) in 31 untreated sarcoidosis patients, 556.6 +/- 341.8 in 10 sarcoidosis patients treated with prednisolone, and 231.5 +/- 156.8 in 16 control individuals. The levels of antibody activities were significantly elevated in untreated patients (p less than 0.05) and in treated patients (p less than 0.02) compared to those of controls. However, considering the treated vs. untreated patients, there was no significant difference in levels. The serum levels of whole immunoglobulin antibody activities were 0.484 +2- 0.191 O.D. in 38 untreated patients, 0.410 +/- 0.166 in 13 treated patients and 0.571 +/- 0.254 in 52 controls. The levels of antibody activity were significantly lower in treated patients than in the controls (p less than 0.05). However, there was no significant difference between the untreated patients and controls. To assess the site of antibody production, the secretion ratio was calculated by dividing the levels in BALF to those in serum. For this purpose, each serum level of antibody activity was also corrected by serum albumin concentration as with BALF.(ABSTRACT TRUNCATED AT 250 WORDS)     

Angiotensin Converting Enzyme in Sarcoidosis and in Silicosis

Angiotensin converting enzyme(ACE) activities of broncho-alveolar lavage fluid(BALF) and serum in patients with sarcoidosis and with silicosis were measured. Serum ACE was measured by enzymic assay and radioimmunoassay. There was a close relationship between ACE activity and content (r=0.78). Serum ACE activities in patients with active sarcoidosis (37.5 ± 11.1 nmol/min/ml, mean ± SD) and with silicosis (25.5 ± 9.3) were significantly elevated from the control (18.6 ± 6.0). BALF ACE activities in the control, patients with active sarcoidosis and with silicosis were 0.23 ± 0.19 nmol/min/ml, 0.94 ± 0.97 and 0.38 ± 0.05, respectively. BALF ACE in patients with active sarcoidosis and with silicosis were significantly different from the control. When BALF ACE was corrected by the cell count of alveolar macrophage (per 10⁶ cells), activity was significantly different from control only in the patients with sarcoidosis. Moreover, only the alveolar macrophages in sarcoidosis were stained by immunofluorescence and immunocytochemistry using rabbit anti-human ACE antibody. Induction of ACE in alveolar macrophage might have an important role for the activity or progression of sarcoidosis.     

Tumor necrosis factor receptors (TNFRs) on T lymphocytes and soluble TNFRs in different clinical courses of sarcoidosis

INTRODUCTION: The release of tumor necrosis factor (TNF-alpha) is increased in sarcoidosis patients. TNF-alpha exerts its effect by binding to specific cell surface receptors. There are only fragmentary data concerning the expression of tumor necrosis factor receptors (TNFRs) on bronchoalveolar lavage fluid (BALF) and peripheral blood (PB) lymphocytes. The aim of the study was to evaluate TNFRI (CD120a) and TNFRII (CD120b) expression on T cells and the level of soluble TNFRs in specimens of patients with different clinical manifestation and clinical outcome of sarcoidosis. MATERIAL AND METHODS: We examined 49 patients with newly diagnosed pulmonary sarcoidosis. TNFRI and TNFRII density on CD4+ and CD8+ BALF and PB cells surface was estimated using monoclonal antibodies and a flow cytometry technique. The level of TNFRs in PB serum and BALF cell culture supernatant (CCS) was measured using ELISA. Immunological analyses were also performed on PB samples collected from 10 healthy volunteers. RESULTS: The level of soluble TNFRI (sTNFRI) in PB serum was similar in sarcoidosis patients and healthy subjects, whereas the concentration of sTNFRII in serum was significantly higher in the sarcoidosis group (P<0.001). Patients without acute symptoms of sarcoidosis, patients with radiological stage II/III as well as patients with further disease progression showed a tendency to higher levels of sTNFRs in PB serum and lower levels of sTNFRs in BALF CCS compared to L?fgren syndrome and radiological stage I subjects, and patients with spontaneous resolution of sarcoidosis. More than 80% of BALF and PB lymphocytes of sarcoidosis patients expressed both CD120a and CD120b antigens. The percentage of double-positive CD4+CD120a+ and CD4+CD120b+ cells in PB was significantly higher (P<0.005) in sarcoidosis patients than in healthy subjects. The highest percentage of CD4+CD120a+ and CD4+CD120b+ lymphocytes in BALF was determined in patients with acute disease, and in PB of patients with further spontaneous improvement. CONCLUSION: The evaluation of sTNFRs and TNFRs expression on T-helper cells may be useful in the estimation of sarcoidosis activity.     

Levels of Transferrin in Bronchoalveolar Lavage Fluid in Sarcoidosis

There has been only one report showing high levels of transferrin (Tf) in bronchoalveolar lavage fluid (BALF) in patients with sarcoidosis. This study was designed to assess the levels of Tf in both BALF and serum and to examine the relationship between the levels of Tf and other disease markers in sarcoidosis. Subjects were 64 sarcoidosis and 10 healthy controls. Tf in BALF and serum was measured by nephelometric assay. Median Tf levels in BALF from sarcoidosis was 0.70 (range, 0.00–3.97) mg/dl, which was significantly higher compared with controls (0.36 (range, 0.00–1.02) mg/dl; p = 0.005). In contrast, median Tf levels in serum from sarcoidosis was 258 (range, 171–383) mg/dl, which was significantly lower compared with controls (322 (range, 234–356) mg/dl; p = 0.003). Tf levels in BALF were significantly correlated with both the percentage of lymphocytes (r = 0.617, p = 0.001) and serum angiotensin-converting enzyme activity (r = 0.363, p = 0.003) and serum soluble interleukin-2 receptor (r = 0.450, p = 0.001) in sarcoidosis. Levels of Tf in BALF from patients with sarcoidosis were not influenced by smoking status. The levels of Tf in sarcoidosis are high in BALF, but low in serum. Increased levels of Tf in BALF may reflect the disease activity.     

Elevated Levels of Type II Soluble Tumor Necrosis Factor Receptors in the Bronchoalveolar Lavage Fluids of Patients with Sarcoidosis

Since tumor necrosis factor (TNF) is known to be involved in granuloma formation in sarcoidosis, and soluble TNF receptors (sTNF-Rs) inhibit TNF action in vivo, we evaluated the levels of sTNF-Rs in the bronchoalveolar lavage fluids (BALF) of 31 subjects using an enzyme-linked immunosorbent assay. Our group consisted of 13 patients with sarcoidosis (7 sarcoidosis patients who received no treatment and 6 who received corticosteroid therapy) and 18 control subjects (11 healthy nonsmokers and 7 asymptomatic smokers). Type II (75-kDa), but not type I (55 kDa) sTNF-R in BALF was elevated significantly in patients with sarcoidosis compared with the healthy nonsmokers (type I: 126.7 ± 17.6 pg/ml BALF vs 79.4 ± 16.5 pg/ml BALF, p > 0.05; type II: 98.3 ± 27.8 pg/ml BALF vs 26.7 ± 4.9 pg/ml BALF, p < 0.05). Although levels of type I sTNF-R in BALF from sarcoidosis patients were not correlated with any cellular profiles of BALF, concentrations of type II correlated significantly with the numbers of lymphocytes in BALF. We concluded that sTNF-R is a normal constituent of the epithelial lining fluids and that levels of type II sTNF-R are elevated significantly in the BALF from individuals with sarcoidosis. This suggests that sTNF-Rs may influence the local bioactivity of TNF and may also contribute to the pathogenesis of sarcoidosis. Accepted for publication: 7 November 1996     

Immunoenzymatic labeling of monoclonal antibodies for surface antigens of T-cells using immune complexes of APAAP in patients with interstitial lung disease]

The use of unlabeled antibody bridging technique with alkaline phosphatase monoclonal anti-alkaline phosphatase (APAAP) complexes makes it possible to solve the problem of short durability of immunofluorescent staining and the problem of nonspecific endogenous enzyme interference of blood cells with immunoperoxidase method. The technique of APAAP allows satisfactorily to demonstrate the cytoplasmic and surface membrane antigens of T-cells both in peripheral blood and bronchoalveolar lavage fluid (BALF). With the technique studied, the subsets of T-lymphocytes simultaneously in both peripheral blood and BALF of 26 patients with interstitial lung disease and of 16 apparently healthy subjects. The results showed: (1) In patients with interstitial pulmonary fibrosis (IPF) CD8 cells in BALF were higher in number than those in peripheral blood and BALF of normal subjects (P < 0.01). It is suggested that abnormalities of T-Lymphocytes might also play a role in the pathogenesis of IPF. (2) CD4 cells in BALF of patients with sarcoidosis were significantly higher in number than those in other groups (P < 0.01). However, CD8 cells in BALF of patients with sarcoidosis were lower in number than those in others (P < 0.01). The higher ratio of CD4/CD8 was found in sarcoidosis patients during active stage. The findings suggested that change of the ratio of CD4/CD8, as an immunoregulatory abnormalities in lung, could be regarded as one of parameters in assessing the activity in patients with sarcoidosis.     

Increased procollagen III aminoterminal peptide-related antigens and fibroblast growth signals in the lungs of patients with idiopathic pulmonary fibrosis

Idiopathic pulmonary fibrosis (IPF) is a chronic lung disorder characterized by an increased density of inflammatory cells, fibroblasts, and collagen within the lung parenchyma. To gain insights into the mechanisms leading to the increased density of fibroblasts and altered collagen metabolism in the IPF lung, bronchoalveolar lavage fluid from normal subjects and patients with IPF or sarcoidosis was analyzed for (1) the presence of antigenic material related to the aminoterminal propeptide domain of type III procollagen, and (2) fibroblast growth-promoting activity in the extracellular milieu of the lower respiratory tract. Whereas bronchoalveolar lavage fluid (BALF) type III procollagen aminoterminal peptide-related antigen levels in 59 patients with sarcoidosis were similar to the levels of control subjects (p greater than 0.10), 31 patients with IPF had markedly increased levels (12-fold over controls; p less than 0.025, IPF versus controls; p less than 0.01, IPF versus sarcoidosis). Type III procollagen aminoterminal peptide-related antigen levels correlated with an increase in the ability of BALF to stimulate fibroblast proliferation (p less than 0.05). Furthermore, BALF from patients with IPF markedly stimulated human lung fibroblast proliferation in vitro (199% increase, p less than 0.01), whereas lavage fluid from patients with sarcoidosis and from control subjects did not. The enhanced fibroblast proliferation induced by IPF BALF occurred in the absence of serum and exogenous growth factors, suggesting that both competence- and progression-type growth factors were present in the lavage fluid.(ABSTRACT TRUNCATED AT 250 WORDS)     

间质性肺疾病患者血清和支气管肺泡灌洗液中内皮素1活？…

评价内皮素对肺纤维化发生，发展作用的影响。方法利用同位素放射免疫直接测定法，检测１０例肺结节病和８例特发性肺纤维化患外周血和支气管肺泡灌洗液中内皮素１（ＥＴ－１）的活性，并与８名健康非吸烟进行对照。结论ＥＴ－１在肺结节病和ＩＰＦ发病机制中起着重要作用，并可作为疾病活动性判定的一项重要参考指标。     

肺结节病支气管肺泡灌洗液中肿瘤坏死因子和中性粒细胞趋化 …

OBJECTIVE: To evaluate the role of cytokines released by the inflammatory cells and immunocytes from patients in pathogenesis of developing sarcoidosis. METHODS: With well- microchemotaxis and allergy immunology, the level of tumor necrosis factor-alpha (TNF-alpha) and neutrophil chemotactic factor (NCF) was measured in the serum and BALF of 11 sarcoidosis and 7 IPF and 8 normal subjects (non-smokers). RESULTS: The levels of TNFalpha (11.9 +/- 3.2, 11.7 +/- 3.0 ng x L(-1)) and NCF (191 +/- 51, 203 +/- 44 cell x 10HP(-1)) of BALF in the patients with sarcoidosis and IPF were significantly higher than these in control group (P < 0.01) and were higher than those in serum. The level of TNF-alpha in the BALF of patients with sarcoidosis was positively correlated with the percentage of lymphocytes (r = 0.73, P < 0.01). The activity of NCF in the BALF of patients with IPF was positively correlated with the percentage of neutrophils (r = 0.89, P < 0.01). CONCLUSIONS: It indicated that TNF-alpha and NCF might play an important role in the pathogenetic process of the sarcoidosis and IPF, and can act as the marker of activity of these diseases.     

Protein profiles of bronchoalveolar lavage fluid from patients with pulmonary sarcoidosis

BACKGROUND: Pulmonary sarcoidosis is a multisystem granulomatous disease with various clinical phenotypes. So far, there has been little information on protein patterns (PPs) of bronchoalveolar lavage fluid (BALF) from patients with sarcoidosis and no data are available on PPs in clinical disease subtypes. OBJECTIVES: To investigate the PP of BALF from patients with pulmonary sarcoidosis, to evaluate whether PPs reflect disease course as assessed by chest X-ray (CXR), and to compare PPs between patients with/without L?fgren's syndrome. METHODS: Surface-enhanced laser desorption/ionization-time-of-flight mass spectroscopy was applied to investigate PPs in unconcentrated BALF from 65 patients (CXR stage I, n = 32; CXR stage II, n = 22, CXR stage III, n = 11) and 23 healthy control subjects. The Mann-Whitney U test was used to detect differentially expressed protein peaks. After reversed-phase fractionation, peptide fingerprint mapping and immunodepletion were used to identify deregulated (up-regulated or down-regulated) proteins. RESULTS: Forty differentially expressed protein entities (2.75-185.62 kD) were detected in patients with pulmonary sarcoidosis versus control subjects (p < 0.05). Whereas 13 peaks (33%) were present across all CXR stages, 27 (67%) were specific for particular CXR stages. Comparison of PPs between CXR stage I patients with or without L?fgren's syndrome revealed 25 differentially expressed peaks. The total number of deregulated peaks and also of those associated with sarcoidosis as a whole were markedly lower in patients with L?fgren's syndrome in comparison with other sarcoid phenotypes. Human serum albumin, alpha1-antitrypsin, and protocadherin-2 precursor were identified from sarcoidosis-associated PP. CONCLUSION: Surface-enhanced laser desorption/ionization-time-of-flight mass spectroscopy enables determination of protein patterns in sarcoid BALF and allows detection of protein patterns linked to a particular disease course.     

Increased endothelin-1 levels of BAL fluid in patients with pulmonary sarcoidosis

OBJECTIVE AND BACKGROUND: Pulmonary fibrosis in sarcoidosis is a significant cause of morbidity and mortality. Various factors have been intensely studied to define the pathogenesis of lung fibrosis in sarcoidosis. Endothelin (ET) consists of three isoforms and is known for its potent vasoconstrictor properties. ET plays an important role in the fibroproliferative process of interstitial lung diseases. METHODS: To investigate the role of ET in the progression of pulmonary fibrosis in sarcoidosis, ET-1 and ET-3 concentrations were measured in BAL fluid (BALF) in 22 non-smoking patients with sarcoidosis and in control subjects (n = 12). Immunoreactivity of ET-1 was also evaluated in alveolar macrophages (AMs) from sarcoidosis patients. To assess the effects of ET in BALF on fibroblast proliferation, human foetal lung fibroblasts were cultured with sarcoidosis or control BALFs in the presence or absence of the ET-receptor antagonist TAK-044. RESULTS: ET-1 levels in sarcoidosis BALF were significantly higher than those in control, whereas ET-3 levels were not different between sarcoidosis and control. ET-1 levels were correlated with the number of AMs in BALF. ET-1-immunoreactivity was found mainly in AM of sarcoidosis BALF. Sarcoidosis BALF significantly stimulated fibroblast proliferation, compared with control BALF, and the fibroblast proliferation induced by sarcoidosis BALF was inhibited by TAK-044. CONCLUSIONS: Increased levels of ET-1 in AM could enhance fibrogenesis in pulmonary sarcoidosis.     

Bronchoalveolar lavage fluid D dimer levels are higher and more prevalent in black patients with pulmonary sarcoidosis

BACKGROUND: Abnormalities of lung coagulation and fibrinolysis in sarcoidosis are thought to play a role in the pathogenesis of this disease. OBJECTIVE: We previously showed that bronchoalveolar lavage fluid (BALF) D dimer directly correlated with various measures of severity in sarcoidosis. Here, we analyze our observation that BALF D dimer was more frequently found at higher levels in African-American patients with pulmonary sarcoidosis. METHODS: BALF D dimer was measured in 55 subjects with pulmonary sarcoidosis and 31 healthy volunteers by enzyme immunoassay. The healthy group established a normal range of BALF D dimer with 71 ng/ml as the highest measured level. This was the cut point for comparisons among the patients with sarcoidosis. RESULTS: High BALF D dimer levels (>71 ng/ml) were found in younger patients with sarcoidosis and were associated with a significantly lower percent predicted forced expiratory volume in 1 s and greater numbers of BAL lymphocytes. Black patients with sarcoidosis had higher BALF D dimer levels (median 131, range 0-2,040 ng/ml) than white patients (median 18, range 0-605 ng/ml; p = 0.011). Higher than normal BALF D dimer levels were found in 61% of the black subjects with sarcoidosis, but in only 20% of the white individuals (chi(2) = 5.539, p = 0.019). BALF D dimer was the only disease measure that discriminated black from white individuals with sarcoidosis. CONCLUSION: BALF D dimer is an indicator of lung fibrin formation and degradation in sarcoidosis. The relationship of high D dimer levels with greater BAL lymphocytosis and worse lung function may be a marker of active sarcoidosis, especially in African-Americans who tend to suffer a more serious form of the disease.     

间质性肺疾病患者血清和支气管肺泡灌洗液中内皮素1活性的变化

目的评价内皮素对肺纤维化发生、发展作用的影响。方法利用同位素放射免疫直接测定法,检测１０例肺结节病和８例特发性肺纤维化（ＩＰＦ）患者外周血和支气管肺泡灌洗液（ＢＡＬＦ）中内皮素１（ＥＴ１）的活性,并与８名健康非吸烟者进行对照。结果肺结节病和ＩＰＦ患者血清和ＢＡＬＦ中的ＥＴ１活性分别为（６２±２９）ｎｇ／Ｌ,（１７０±２４）ｎｇ／Ｌ和（７７±７１）ｎｇ／Ｌ、（１０±３）ｎｇ／Ｌ,与正常对照组（２０±８）ｎｇ／Ｌ、（４０±０６）ｎｇ／Ｌ比较,差异有显著性（Ｐ＜００１）;血清中ＥＴ１活性与动脉血氧分压（ＰａＯ２）呈明显负相关（ｒ＝－０５３８,Ｐ＜００１）;结节病组和ＩＰＦ组ＢＡＬＦ中的ＥＴ１水平与ＢＡＬＦ中细胞总数呈正相关（ｒ＝０６４９,Ｐ＜００１）,肺结节病患者、ＩＰＦ患者ＢＡＬＦ中ＥＴ１与淋巴细胞、中性粒细胞呈正相关（ｒ＝０７１２,０８１３,Ｐ均＜００１）。结论ＥＴ１在肺结节病和ＩＰＦ发病机制中起着重要作用,并可作为疾病活动性判定的一项重要参考指标。     

Different angiogenic activity in pulmonary sarcoidosis and idiopathic pulmonary fibrosis

BACKGROUND: Recent evidence has shown that several chemokines--including those involved in angiogenesis--have been implicated in the pathogenesis of idiopathic pulmonary fibrosis (IPF) and sarcoidosis. We speculated that these differences could be attributed to distinct angiogenic and angiostatic profiles. This hypothesis was investigated by estimating the levels of three angiogenic chemokines (growth-related gene [GRO]-alpha, epithelial neutrophil-activating protein [ENA]-78, and interleukin [IL]-8), and three angiostatic chemokines (monokine induced by interferon (IFN)-gamma [MIG], IFN-gamma-inducible protein [IP]-10, and IFN-gamma-inducible T-cell alpha chemoattractant) in serum and BAL fluid (BALF). METHODS: We studied prospectively 20 patients with sarcoidosis (median age, 46 years; range, 25 to 65 years), 20 patients with IPF (median age, 68 years; range, 40 to 75 years), and 10 normal subjects (median age, 39 years; range, 26 to 60 years). RESULTS: The GRO-a serum and BALF levels of IPF patients were found significantly increased in comparison with healthy subjects (799 pg/mL vs 294 pg/mL [p = 0.022] and 1,827 pg/mL vs 94 pg/mL [p < 0.001], respectively) and sarcoidosis patients (799 pg/mL vs 44 pg/mL [p < 0.001] and 1,827 pg/mL vs 214 pg/mL [p < 0.001], respectively). Moreover, ENA-78 and IL-8 BALF levels in IPF patients were significantly higher compared with sarcoidosis patients (191 pg/mL vs 30 pg/mL [p < 0.001] and 640 pg/mL vs 94 pg/mL [p = 0.03], respectively). MIG serum levels in IPF patients were found significantly upregulated in comparison with sarcoidosis patients and healthy control subjects. However, MIG and IP-10 BALF levels (1,136 pg/mL vs 66 pg/mL [p < 0.001] and 112 pg/mL vs 56 pg/mL [p = 0.037], respectively) were significantly higher in sarcoidosis patients compared with IPF patients. CONCLUSIONS: Our data suggest distinct angiogenic profiles between IPF and sarcoidosis, indicating a potential different role of CXC chemokines in the local immunologic response in IPF and pulmonary sarcoidosis.     

The clinical significance of mast cell tryptase in bronchial alveolar lavage fluid in interstitial lung diseases]

Mast cell tryptase plays an important role in fibrosis. Tryptase levels in bronchial alveolar lavage fluid (BALF) from patients with interstitial lung diseases are frequently increased, but little is known of the clinical significance. The study population consisted of 93 patients [38 with sarcoidosis, 23 with collagen vascular disease (CVD), and 32 with idiopathic pulmonary fibrosis (IPF)]. BALF tryptase levels were measured with a newly developed B12 antibody-fluoroimmunocap method (UniCAP method), which can detect an activated form of tryptase. We examined the relationship between BALF tryptase levels and clinical parameters of the diseases. BALF tryptase was detected in 7 (18.4%) patients with sarcoidosis, 7 (30.4%) with CVD, and 14 (45.8%) with IPF. In tryptase-positive group, serum ACE levels and the numbers of BALF-mast cells and lymphocytes were higher than the tryptase-negative group in sarcoidosis, serum LDH levels were higher in CVD, and the number of BALF-lymphocyte and Hugh-Jones grade were higher in IPF. Furthermore, tryptase-positive IPF cases had a poorer outcome than the tryptase-negative group by Kaplan-Meier analysis. Tryptase in BALF detected with the UniCAP method may be associated with disease activity in sarcoidosis and CVD, and with severity and poor prognosis in IPF. BALF tryptase measurement may be useful in the assessment of disease activity and severity in various interstitial lung diseases.     

Increased levels of nerve growth factor in the airways of patients with sarcoidosis

Objectives. Nerve growth factor (NGF) is a potent neuronal growth factor with inflammatory properties that recently has been proposed to be of importance in airway pathology. A role for NGF in the inflammatory granulomatous lung disease sarcoidosis is not well elucidated. The aims of this study were to investigate the secreted levels of NGF in bronchoalveolar lavage fluid (BALF) from sarcoidosis patients compared with patients with resolved disease, patients with another granulomatous disease – chronic beryllium disease (CBD) – and healthy subjects and also to investigate the relationship between NGF levels and markers of inflammation. Methods and results. NGF levels in BALF from 56 patients with active sarcoidosis (22 with Löfgren’s syndrome), nine subjects with resolved sarcoidosis, six patients with CBD, and 31 healthy subjects were compared. A 10‐fold elevation of NGF levels was found in patients with active sarcoidosis compared with subjects with clinically resolved sarcoidosis, patients with CBD and healthy subjects. In sarcoidosis patients, positive correlations between concentrations of NGF and lymphocytes, eosinophils and interferon‐γ, interleukin (IL)‐4, IL‐10, IL‐12 were found. Conclusions. We demonstrate that secreted levels of NGF are markedly enhanced in the airways in active pulmonary sarcoidosis. Furthermore, a relationship between NGF and pulmonary inflammation in sarcoidosis is supported.     

间质性肺疾病支气管肺泡灌洗液中性粒细胞趋化因子和肿瘤坏 …

OBJECTIVE: To investigate the relationship between the level of the neutrophil chemotactic factor(NCF), tumor necrosis factor-alpha(TNF-alpha) in patients with interstitial lung disease(ILD) and the activity of ILD. METHOD: The NCF activities in the BALF and in the serum from 11 patients with sarcoidosis, 7 with idiopathic pulmonary fibrosis (IPF) and 8 normal subjects were determined using the membrane filter and radio-immunoassay. The level of TNF-alpha was also detected. RESULT: In the 7 IPF patients, the level of NCF and TNF-alpha (203 +/- 44 cells/10 HP, 11.7 +/- 2.9 ng/L) in the BALF was higher than that in 8 control patients (83 +/- 45 cells/10 HP, 6.5 +/- 1.4 ng/L, P < 0.01). The level of NCF and TNF-alpha in the BALF from 11 patients with sarcoidosis (186 +/- 50 cells/10 HP, 12 +/- 3 ng/L) was highet than those in 8 control patients (P < 0.01). The level of NCF and TNF-alpha in the BALF from patients with IPF was positive correlated with the percentage of neutrophil (NCF: r = 0.89, P < 0.01; TNF-alpha: r = 0.86, P < 0.05). The level of NCF and TNF-alpha in the BALF of patients with sarcoidosis was positive correlated with the percentage of lymphocyte (NCF: r = 0.78, P < 0.01; TNF-alpha: r = 0.73, P < 0.01. CONCLUSION: The level of NCF and TNF-alpha in the BALF from patients with IPF and sarcoidosis can act as the marker of the activity of alveolitis of IPF and sarcoidosis.     

T‐cell phenotypes in bronchoalveolar lavage fluid,blood and lymph nodes in pulmonary sarcoidosis – indication for an airborne antigen as the triggering factor in sarcoidosis

Abstract. Darlington P, Haugom‐Olsen H, von Sivers K, Wahlström J, Runold M, Svjatoha V, Porwit A, Eklund A, Grunewald J (Karolinska Institutet, Stockholm, Sweden). T‐cell phenotypes in bronchoalveolar lavage fluid, blood and lymph nodes in pulmonary sarcoidosis – indication for an airborne antigen as the triggering factor in sarcoidosis. J Intern Med 2012; 272: 465–471. Background. An increased percentage of CD4+ T cells is usually observed in bronchoalveolar lavage fluid (BALF) from patients with sarcoidosis. In HLA‐DRB1*03‐positive patients, such T cells express the T‐cell receptor (TCR) AV2S3+ gene segment. It is not known whether cells found in BALF reflect those in enlarged regional lymph nodes (LNs). Therefore, the aim of this study was to compare T‐cell phenotypes in BALF, blood and mediastinal LNs. Methods. Fifteen patients underwent clinical investigation including bronchoscopy with bronchoalveolar lavage. Blood samples were drawn, and endoscopic ultrasound‐guided fine‐needle aspiration of enlarged mediastinal LNs was performed via the oesophagus. T cells from all three compartments were analysed by flow cytometry for markers of activity, differentiation and T regulatory function. Results. The CD4/CD8 ratio was significantly higher in BALF compared with regional LNs and was also significantly higher in LNs than in blood. The CD4+ T cells were recently activated and more differentiated in BALF than in blood and LNs. There was an accumulation of T regulatory cells (FOXP3+) in LNs and a correlation between high levels of FOXP3+ cells in BALF and in LNs. In HLA‐DRB1*03‐positive patients, TCR AV2S3+ CD4+ T cells were predominantly localized within BALF. Conclusions. The CD4+ T‐cell phenotype in BALF indicates an active ongoing specific immune response primarily localized to the alveolar space.     

Perforin down-regulation and adhesion molecules activation in pulmonary sarcoidosis: an induced sputum and BAL study

BACKGROUND: Sarcoidosis is thought to be a T-helper type 1 cytokine-mediated disorder. Sputum induction has been proposed as a useful noninvasive method mainly for the assessment of airway diseases. However, it is unknown whether the balance of T-cytotoxic (Tc1) type 1 and Tc2 cells is altered in sarcoidosis. STUDY OBJECTIVES: The primary aim of this study was to characterize the CD8+ T lymphocyte subpopulations in induced sputum from sarcoidosis patients, and to compare these subpopulations to those found in BAL fluid (BALF) from sarcoidosis patients. To further investigate the mechanism of the cytotoxic activity of CD8+ lymphocytes, we measured their perforin expression. Additionally, two adhesion molecules (CD62 and CD71), which are expressed on CD8+ T cells and may serve as novel immunologic markers, were detected. Settings: Department of Thoracic Medicine, University of Crete, and Department of Pneumonology, Democritus University of Thrace, Alexandroupolis, Greece. PATIENTS: We prospectively studied 22 patients with sarcoidosis (median age, 48 years; age range, 25 to 65 years) and 10 healthy subjects (5 female and 5 male; median age, 39 years; age range, 26 to 60 years). INTERVENTIONS: The stimulation of lymphocytes with phorbol 12-myristate 13-acetate was followed by the use of double immunocytochemical methods to identify CD8+ interferon (IFN)-gamma producing cells (ie, Tc1) and CD8+ interleukin-4 producing cells (ie, Tc2). MEASUREMENTS AND RESULTS: We found a significant decrease in the prestimulation percentage of IFN-gamma-positive CD8+ T cells in the BALF (p = 0.001) and induced sputum (p = 0.001) of sarcoidosis patients compared to the number in samples from healthy control subjects. However, no significant difference was documented between lymphocyte subsets poststimulation. Decreased levels of perforin expression were found in BALF (p = 0.001) and induced sputum (p < 0.001) of sarcoidosis patients compared to those in control subjects. The adhesion molecules were significantly increased in both the BALF and induced sputum of the sarcoid population compared to those in healthy control subjects. CONCLUSIONS: Our results suggest that inflammation could be effectively and noninvasively determined by using sputum induction in sarcoidosis patients. In addition, we have provided evidence suggesting the possibility that CD8+ lymphocytes might not play a major role in sarcoidosis.