Immunoglobulins and cellular constituents of the BAL fluid of patients with sulfur mustard gas-induced pulmonary fibrosis.

STUDY OBJECTIVE: The acute heavy exposure to sulfur mustard gas can lead to pulmonary fibrosis (PF). This study was performed to determine the cellular and protein content of BAL fluid in 24 patients with sulfur mustard gas-induced PF. PATIENTS: Twenty-four veterans with sulfur mustard gas-induced PF and 18 nonexposed veterans serving as control subjects were enrolled into the study. MEASUREMENTS: Chest roentgenograms, pulmonary function tests (PFTs), tests for carbon monoxide diffusing capacity of the lung (DLCO), high-resolution CT scans of the chest, BAL via fiberoptic bronchoscopy, analyses of BAL fluids for cellular and protein constituents, and determinations of serum albumin and Ig levels were performed in all cases. A transbronchial lung biopsy was done in all patients following BAL. RESULTS: Neutrophilic alveolitis was the predominant feature. Neutrophils (p = 0.0001) and eosinophils (p = 0.0001) were the predominant cell types in the BAL fluid of patients with PF. There was a strong correlation between the BAL fluid neutrophil count (p = 0.76; p = 0.0003) or its percentage (p = 0.77; p = 0.0003) and the degree of fibrosis. Of the BAL fluid Ig levels, only the IgG level in the study group was significantly higher than the IgG level of the control group (p = 0.0001). Of the PFT physiologic parameters, only the percentage of DLCO showed a significant correlation with the degree of fibrosis (p = -0.76; p < 0.001). CONCLUSION: The cellular constituents of BAL fluid in patients with sulfur mustard gas-induced PF are very similar to the cellular constituents seen in patients with idiopathic PF, and this finding indicates the presence of an ongoing active alveolitis in PF.     

Characteristics of bronchoalveolar lavage fluid in patients with sulfur mustard gas-induced asthma or chronic bronchitis.

PURPOSE: To examine the pattern of immunoglobulins and cellular constituents in bronchoalveolar lavage fluid obtained from patients with sulfur mustard gas-induced asthma or chronic bronchitis as compared with healthy control subjects. SUBJECTS AND METHODS: We studied two groups of nonsmoking veterans with either bronchial asthma (n = 21) or chronic bronchitis (n = 28) believed to have been caused by sulfur mustard gas exposure and a third group of healthy, nonsmoking, non-sulfur mustard gas exposed controls (n = 17). Bronchoalveolar lavage was performed in all three groups. The cellular constituents, albumin content, and immunoglobulin concentrations were determined. RESULTS: The three groups did not differ in age or in the serum albumin and immunoglobulin concentrations. The volume of bronchoalveolar lavage fluid recovered was approximately 10% less in the patients with asthma and chronic bronchitis (P = 0.008). The proportions of lymphocytes among the bronchoalveolar lavage cells were similar in all three groups, whereas the proportion of eosinophils was greater in lavage fluid from the asthmatic subjects than in either the healthy control subjects or the patients with chronic bronchitis (P = 0.0001). Both the total number of the recovered cells per milliliter of lavage fluid and the proportion of neutrophils were significantly greater in bronchoalveolar lavage from patients with chronic bronchitis than in healthy subjects or in the patients with asthma (all P <0.001). CONCLUSION: The bronchoalveolar lavage cellular constituents of patients with sulfur mustard gas-induced asthma and chronic bronchitis are similar to those that have been observed previously in patients with asthma and chronic bronchitis from other common causes.     

Shifts of T4/T8 T lymphocytes from BAL fluid and peripheral blood by clinical grade in patients with pulmonary tuberculosis

OBJECTIVES: We investigated the shifts of T4/T8 lymphocytes from BAL fluid (BALF) and peripheral blood by the clinical grade of pulmonary tuberculosis (TB), which is determined by factors such as extent of pulmonary involvement, fever, and loss of body weight. MATERIALS AND METHODS: In order to explore these questions, BALF was collected from 45 patients presenting with active pulmonary TB and 14 healthy control subjects. The percentages for T-lymphocyte subpopulations, including CD4(+), CD8(+), and CD3(+) T cells, were measured using two-color flow cytometry. RESULTS: A higher percentage of CD3(+)CD4(+) T lymphocytes, with a relatively lower percentage of CD3(+)CD8(+) T lymphocytes, was revealed for the patients with a higher grade of pulmonary TB, compared to patients with a lower grade of pulmonary TB, resulting in an increased BALF C4(+)/CD8(+) ratio. By contrast, a higher percentage of CD3(+)CD8(+) T lymphocytes with a relatively low percentage of CD3(+)CD4(+) T lymphocytes was demonstrated for these patients with a higher grade of pulmonary TB, resulting in a decreased peripheral blood CD4(+)/CD8(+) ratio. CONCLUSIONS: Our findings suggest that compartmentalization of the CD4(+) T lymphocytes in the infected lungs may occur for patients with higher grades of pulmonary TB.     

Relationship of BAL and lung tissue CD4+ and CD8+ T lymphocytes, and their ratio in idiopathic pulmonary fibrosis

BACKGROUND: Surgical biopsy specimens have shown that T lymphocytes (TLs) infiltrate lung parenchyma in patients with idiopathic pulmonary fibrosis (IPF) and might play a pathogenetic role. BAL, a far less invasive technique, has also been used for the investigation of IPF pathogenesis. However, controversy exists whether the BAL fluid cellular profile reflects the cellular composition of the lung parenchyma. STUDY OBJECTIVE: To compare infiltrating TLs subpopulations (CD4+, CD8+, and CD4+/CD8+ ratio) in lung tissue and BAL fluid. PATIENTS AND METHODS: Immunohistochemistry was performed according to the streptavidin-biotin method on the surgical biopsy specimens of 12 untreated patients with IPF. The number of CD3+, CD4+, and CD8+ TLs was determined by observer-interactive computerized image analysis (SAMBA microscopic image processor; Meylan, France). In BAL fluid, the same TLs subpopulations were evaluated by flow cytometry. RESULTS: In lung tissue, CD3+ TLs accounted for a mean (+/- SEM) of 28.8 +/- 7% of total cells, CD4+ TLs accounted for 14.5 +/- 4% of total cells (50.1 +/- 4% of CD3+ TLs), and CD8+ TLs accounted for 13.8 +/- 4% of total cells (47.4 +/- 4% of CD3+ TLs). In BAL fluid, lymphocytes accounted for 9.8 +/- 2.5% of total cells, CD4+ TLs accounted for 51.8 +/- 4% of CD3+ TLs, and CD8+ TLs accounted for 42.2 +/- 4% of CD3+ TLs. Tissue CD4+ and CD8+ TLs (expressed as a percentage of CD3+ TLs) correlated significantly with the number of CD4+ and CD8+ TLs in BAL fluid (r = 0.846 and p = 0.001 vs r = 0.692 and p = 0.013, respectively). A significant positive correlation was also found between the mean CD4+/CD8+ ratio found in tissue and BAL fluid (1.05 +/- 0.21 and 1.5 +/- 0.27, respectively; r = 0.832; p = 0.01). CONCLUSION: The results suggest that in patients with IPF, the TL subpopulations in BAL fluid reflect the pattern of lymphocytic infiltration in pulmonary parenchyma.     

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.     

Frequency of telomerase-specific CD8+ T lymphocytes in patients with cancer

Telomerase is considered a universal tumor-associated antigen (TAA) due to its high rate of expression by cancers (approximately 90%), and clinical trials are in progress to test the immunotherapeutical efficacy of antitelomerase immunization in patients with cancer. However, the data concerning frequency and functional activity of telomerase-specific cytotoxic T lymphocytes (CTLs) in patients with cancer are few and conflicting, although their knowledge would be mandatory to predict the efficacy of telomerase-specific immunotherapy in selected patients. We performed this study to analyze frequency and cytolytic function of circulating CD8+ T lymphocytes specific for the p540 telomerase peptide in a series of human leukocyte antigen (HLA)-A2+ cancer patients. The results show that most patients with cancer have circulating telomerase-specific CD8+ T lymphocytes, but a high frequency of telomerase-specific CTLs are present only in a fraction of them. Furthermore, CTL lines able to kill telomerase-positive tumor cells, including autologous cancer cells, can be expanded ex vivo from some, but not all, patients with cancer. In conclusion, the results of the study support the development of clinical protocols using telomerase peptides as an immunizing agent. However, they underline the necessity to study single patients immunologically before undergoing vaccination, to select the patients adequately, and to eventually adapt the immunization schedule to the patient's immunologic status.     

Diagnostic role of BAL fluid CD4/CD8 ratio in different radiographic and clinical forms of pulmonary sarcoidosis

Introduction: Bronchoalveolar lavage (BAL) as a method of sampling cells is useful in the diagnosis and differential diagnosis of sarcoidosis. However, CD4/CD8 ratio in BAL fluid (BALF) is highly variable and it generates continuous discussions about its diagnostic role. Objective: To prospectively evaluate diagnostic role of BALF CD4/CD8 ratio in pulmonary sarcoidosis manifested in different radiographic and clinical forms in the real clinical practice. Material and methods: The study population consisted of 318 sarcoid patients with a newly diagnosed disease. Comparator groups consisted of 55 healthy subjects and 130 patients with other disorders who underwent BAL and examination of CD4/CD8 ratio in BALF as a step of diagnostic pathway. Diagnostic accuracy of CD4/CD8 ratio in BALF using receiver‐operating characteristic analysis has been calculated. Results: The percentage of BALF lymphocytes in sarcoid patients was significantly different from comparator groups. Normal BALF cell counts were found in 7% of sarcoid patients. However, typical sarcoid BALF cellular pattern was found in 6.2% of all control subjects. We have found that optimal cutoff points for CD4/CD8 ratio are 3.5 and 4.0 for asymptomatic and symptomatic patients, respectively. Sensitivity of the optimal cutoff points of CD4/CD8 ratio was lower in asymptomatic patients compared with symptomatic patients. Sensitivity of the optimal cutoff points decreased with the increased stage of sarcoidosis. Conclusions: BAL is a valuable method in diagnostic pathway of pulmonary sarcoidosis. However, results of BALF examination must be interpreted considering a specific clinical case. BALF CD4/CD8 ratio depends on clinical and radiographic manifestation. Please cite this paper as: Danila E, Norkūnien? J, Jurgauskien? L and Malickait? R. Diagnostic role of BAL fluid CD4/CD8 ratio in different radiographic and clinical forms of pulmonary sarcoidosis. The Clinical Respiratory Journal 2009; 3: 214–221.     

肺结核患者CD4、CD8淋巴细胞变化分析

目的　观察肺结核患者CD4、CD8淋巴细胞变化,探讨其免疫状态的改变及其意义。方法 用流式细胞仪测定31例成人肺结核及12例对照组的CD4、CD8计数值,对测定结果进行统计学分析。结果 病人组CD4值明显下降,CD8值无明显变化,CD4下降幅度与病情严重程度、排菌状态无明显关联。结论 成人肺结核患者存在以CD4T淋巴细胞下降为突出表现的细胞免疫受损,但其对病情发生、发展无独立的影响作用。     

High CD95 expression of BAL lymphocytes predicts chronic course in patients with sarcoidosis

BACKGROUND AND OBJECTIVES: The prognosis of sarcoidosis is highly variable, with spontaneous remission in some patients. Apoptosis may be associated with spontaneous resolution of the granulomata. CD95 (Fas), an apoptotic molecule, and CD29 and CD45RO (T-cell memory markers) are expressed at higher levels on T lymphocytes from sarcoid patients compared with normal subjects. However, the prognostic significance of CD95, CD29 and CD45RO expression in sarcoidosis is not clear. It was hypothesized that expression of CD95 would correlate with spontaneous remission. METHODS: CD29, CD45, CD45RO and CD95 expression of BAL fluid and peripheral blood (PB) lymphocytes was studied with flow cytometry in 50 patients with sarcoidosis. Results of the 15 chronic patients and 21 patients who remitted spontaneously were compared. RESULTS: BAL CD95 (59 vs 8, P = 0.002), and PB CD95 (48 vs 18, P = 0.004) and PB CD45RO (50 vs 41, P = 0.003) expression was significantly higher in patients with chronic disease compared with those with spontaneous remission. Sensitivity, specificity, positive predictive value, negative predictive value and accuracy for these markers were: BAL CD95 (cut-off: 42.5%) 73.3%, 85.7%, 78.6%, 81.8% and 80.6%; PB CD95 (cut-off: 25%) 86.7%, 66.7%, 65%, 87.5% and 75%; and PB CD45RO (cut-off: 44.5%) 80%, 61.9%, 60%, 81.3% and 69.4%, respectively. CONCLUSION: Levels of BAL and PB CD95 and PB CD45RO were unexpectedly elevated in patients with chronic disease and may be useful in predicting prognosis in patients with sarcoidosis. Further studies with more patients are necessary to confirm the prognostic role and cut-off value for these markers.     

Soluble form of fas and fas ligand in BAL fluid from patients with pulmonary fibrosis and bronchiolitis obliterans organizing pneumonia

STUDY OBJECTIVES: The Fas-Fas ligand (FasL) pathway is a representative system of apoptosis-signaling receptor molecules. We previously described that this pathway may play an important role in the pathogenesis of fibrosing lung diseases. In this study, we hypothesized that soluble form of Fas (sFas) and FasL (sFasL) may also be associated with this disorder. MEASUREMENTS AND RESULTS: We measured sFas and sFasL levels in BAL fluid (BALF) from patients with idiopathic pulmonary fibrosis (IPF), interstitial pneumonia associated with collagen vascular diseases (CVD-IP), and bronchiolitis obliterans organizing pneumonia (BOOP), using enzyme-linked immunosorbent assay. BALF from all patients was obtained before prednisolone therapy. sFasL levels were relatively increased in IPF patients (p = 0.084), and significantly increased in CVD-IP patients (p < 0.05) and BOOP patients (p < 0.05), compared with control subjects. BALF sFasL levels were elevated in the IPF or CVD-IP subgroups with an indication for prednisolone therapy, compared with those without an indication for therapy. The BALF sFasL level in IPF patients was correlated with the number of total cells and lymphocytes. The BALF sFasL level in BOOP patients was relatively or significantly correlated with the number of total cells or lymphocytes, respectively. The BALF sFas level was significantly increased in BOOP patients, but not in IPF or CVD-IP patients. CONCLUSIONS: We conclude that BALF sFasL levels may be associated with the accumulation of inflammatory cells and reflect the degree of lymphocyte alveolitis in IPF. The elevation of sFasL may be associated with the deterioration of IPF and CVD-IP. The elevation of the BALF sFas level may abrogate the cytotoxicity of FasL in BOOP patients, which may be associated with better prognosis of BOOP, compared with IPF or CVD-IP.     

CD+8CD-28T淋巴细胞在肺结核发病机制中的作用

目的 探讨CD8^＋CD28^-T淋巴细胞在肺结核发病机制中的作用。方法 30例病例为2005年3月至5月遵义医学院附属医院呼吸内科住院及门诊患者，采用流式细胞术检测15例肺结核组患者外周血中CD8^＋CD28^-T淋巴细胞比值、细胞内白细胞介素6（IL-6）水平，以及CD3^＋、CD3^＋CD8^＋、CD8^CD28^＋T淋巴细胞比值，15例慢性支气管炎急性发作期患者作为疾病对照组，15名健康人作为健康对照组。结果 肺结核组和疾病对照组CD3^＋T淋巴细胞分别为（41±16）％和（40±10）％，均显著低于健康对照组[（44±6）％]；肺结核组和疾病对照组CD8^＋CD28^＋T淋巴细胞[（47±16）％和（44±10）％]均显著高于健康对照组[（41±12）％]；肺结核组CD8^＋CD28^＋T淋巴细胞[（15±8）％]显著低于疾病对照组[（20±7）％]，两组均显著低于健康对照组[（32±9）％]；肺结核组CD8^＋CD28^-T淋巴细胞[（27±9）％]显著高于疾病对照组[（22±9）％]，两组均显著高于健康对照组[（10±4）％]；肺结核组CD8^＋CD28^-T淋巴细胞分泌的IL-6水平[（32．4±2．4）％]显著高于疾病对照组和健康对照组[（19．7±3．2）％和（15．2±2．7）％]。结论 肺结核患者CD8^＋CD28^-T淋巴细胞及其分泌的IL-6水平在外周血中上调，CD8^＋CD28^＋T淋巴细胞水平在外周血中下调。CD8^＋CD28^＋和CD8^＋CD28^-T淋巴细胞及其分泌的IL-6可能参与肺结核的发病机制。CD8^＋CD28^＋未和CD8^＋CD28^-T淋巴细胞比值及其分泌的IL-6水平可作为活动性肺结核的辅助诊断指标。     

Increased levels of oxidized methionine residues in bronchoalveolar lavage fluid proteins from patients with idiopathic pulmonary fibrosis

Phagocytic cells are believed to play a crucial role in the development of inflammatory lung diseases. We assumed that the oxidation of methionine (met) to methionine sulfoxide [met(O)] by oxygen-derived free radicals released from phagocytes is one parameter to identify the oxidative mechanisms of lung injury. To test this hypothesis we determined the molar ratio of met(O)/met in the soluble protein fraction of bronchoalveolar lavage (BAL) fluids from healthy nonsmokers and from nonsmoking patients with idiopathic pulmonary fibrosis (IPF) or sarcoidosis. The met(O)/met ratio of the healthy nonsmoker group (n = 11) was 0.046 +/- 0.008 (mean +/- SEM). In contrast, the met(O)/met ratio of the nonsmoking IPF group (n = 11) was significantly increased to 0.223 +/- 0.053 (p less than 0.0002). The BAL fluids of this group showed strongly increased numbers of neutrophils but normal numbers of alveolar macrophages (AM). In the sarcoidosis group (n = 10) the met(O)/met ratio (0.048 +/- 0.010) was not significantly different from control values. A close relationship was found between the met(O)/met ratios and the relative as well as the absolute neutrophil counts (r = 0.86; p less than 0.0002; n = 22). In contrast, no significant correlation was found between the met(O)/met ratios and the absolute AM counts (r = 0.22; p = 0.32; n = 22). We conclude that mechanisms of oxidative lung injury in IPF can be characterized by oxidation of met and that this oxidation may be mediated by neutrophils.     

Human immunodeficiency virus interactions with CD8+ T lymphocytes

Human immunodeficiency virus (HIV)-specific CD8+ T cells can mediate anti-HIV activity by both cytolytic (cytotoxic T lymphocyte or CTL) and non-cytolytic mechanisms (antiviral) and play a crucial role in HIV pathogenesis. Both mechanisms actively contribute to the control of HIV in vivo. The non-cytolytic CD8+T cells from individuals infected with HIV suppress virus replication in CD4+ T cells in vitro by a non-cytolytic mechanism that involves interplay of several chemokines and an unidentified secreted soluble CD8 (+)-cell antiviral factor (CAF). There is immense value of these two distinct CD8 activities in anti-HIV responses and their necessity to be maintained during highly active antiretroviral therapy (HAART). The aim of this review is to provide an overview of some of the novel aspects of CD8+ T cell interactions with HIV, their role in HIV pathogenesis, HAART therapy, HIV disease progression, gene expression and interactions with other cell types during HIV infection.     

Elevated levels of alpha-defensins in plasma and BAL fluid of patients with active pulmonary tuberculosis

STUDY OBJECTIVES: To investigate the role of neutrophil peptides named alpha-defensins in patients with pulmonary tuberculosis (TB). PATIENTS: Thirty-seven patients with TB and 25 healthy subjects. MEASUREMENTS AND RESULTS: Concentrations of alpha-defensins (human neutrophil peptide [HNP]-1, HNP-2, and HNP-3) were measured by radioimmunoassay in plasma and BAL fluid (BALF). Concentrations of alpha-defensins were significantly higher in plasma and BALF of patients with TB than in healthy subjects. In BALF of patients with TB, the concentration of alpha-defensins correlated positively with the levels of interleukin 8, and higher concentrations of alpha-defensins in BALF were also detected in patients with cavitary lesions. There was an inverse relationship between plasma alpha-defensins and FEV(1)/FVC ratio before treatment, and between plasma concentrations of alpha-defensins before treatment and the improvement in percentage of vital capacity after treatment. Plasma alpha-defensin concentrations returned to the normal range after treatment. CONCLUSION: Our data suggest that alpha-defensins released from neutrophils may play an important role in the pathogenesis of TB, and that plasma alpha-defensin concentration may be a useful marker of disease severity and deterioration of pulmonary function.