大鼠哮喘模型外周血和支气管肺泡灌洗液中γδ T细胞明显活化

Ｔ淋巴细胞依据其表面的细胞抗原受体 (ＴＣＲ)的不同 ,分为αβＴ细胞和γδＴ细胞。气道淋巴细胞和嗜酸粒细胞的浸润是支气管哮喘的主要病理特征 ,但尚未见有关γδＴ细胞在哮喘中变化的研究报道。我们探讨了大鼠哮喘模型外周血和支气管肺泡灌洗液 (ＢＡＬＦ)中γδＴ细胞亚群的数量变化和活性改变。采用雄性Ｗｉｓｔａｒ大鼠 ,共 2 0只 ,随机分哮喘模型组和正常对照组各 10只 ;模型组应用鸡卵清蛋白 (ＯＶＡ)注射致敏和雾化吸入刺激 ,制作致敏大鼠哮喘模型 ;对照组以磷酸盐缓冲液代替ＯＶＡ注射致敏和雾化吸入。以激发反应、ＢＡＬＦ细胞…     

哮喘患者动脉血及支气管肺泡腔灌洗液中内皮素—1的浓度

内皮素是一个具有很多生物学特性的缩氨酸介质家族。其中包括收缩气道和血管的作用。内皮素 - 1(ET- 1)比同家族的其余两种缩氨酸更有效。为了了解 ET- 1在哮喘发病机理中的作用 ,我们检测了 11位遗传性过敏性哮喘患者发作期和缓解期以及 11位健康对照者的动脉血中 ET- 1水平 ,并通过纤维支气管镜对 11位缓解期哮喘患者和 11位健康对照者行支气管灌洗术以检测支气肺泡腔灌洗液 (BAL )中 ET- 1水平。动脉血及 BAL中的 ET- 1浓度用放免法测得。动脉血 ET- 1水平不论在哮喘持续组 (3.6 7± 0 .5 1pg/ml)或缓解组 (2 .6 5± 10 .6 2 pg/…     

支气管哮喘小白鼠支气管肺泡灌洗液中TGF-β含量的变化

气道重构 (airwayremodeling)是难治性支气管哮喘气道发生不可逆性气流阻塞及持续性非特异性气道高反应性的重要原因之一。TGF β是分子量约 2 5kD的蛋白质 ,最近报道TGF β具有刺激纤维母细胞促进纤维结合素及胶原的等间质成分的产生[1 ] ,于是推测TGF β在支气管哮喘气道重构中可能起重要的作用。在国内定量性研究TGF β在支气管哮喘气道重构中的作用尚无文献报道。本实验用小白鼠建立支气管哮喘气道重构的动物模型测定 (bronchoalveollarlavageflu id ,BALF)中TGF β的含量阐明气道重构中它的作用。1　材料与方法1 1　动物 :动物…     

选择性扩增外周血和支气管肺泡灌洗液中γδT细胞的研究

目的 :探讨选择性扩增外周血和支气管肺泡灌洗液 (BALF)中γδT细胞的方法 ,以获得高纯度的γδT细胞亚群。方法 :应用梯度离心法分离大鼠 (n =10 )外周血和BALF中的单个核细胞 ,经贴壁除去单核 /巨噬细胞后 ,用补体攻击αβT细胞 ,再用抗TCRγδ单克隆抗体 (mAb)通过固相法加IL 2刺激选择性培养扩增γδT细胞 (简称“攻击洗淘法”) ;通过细胞生长曲线观察细胞增殖变化 ,采用免疫组化法和流式细胞仪检测鉴定γδT细胞纯度。结果 :PBMC和BALF中的γδT细胞在抗TCRγδmAb和IL 2存在下 ,无须再用抗原刺激 ,能维持较长时间增殖 ;经“攻击洗淘法”纯化后扩增的γδT细胞纯度达到 81%～99%。结论 :“攻击洗淘法”能高度纯化扩增外周血和BALF中的γδT细胞     

支气管哮喘病人T淋巴细胞亚群测定

本文应用抗人T淋巴细胞亚群单克隆抗体测定了90例哮喘儿童及32例正常对照儿童外周血T淋巴细胞亚群,结果表明哮喘病人无明显成熟T淋巴细胞、T_H亚群和T_s亚群异常。提示病人B淋巴细胞过度产生IgE可能是在更精细的水平上受T淋巴细胞所调节。     

人类T_H细胞亚群与支气管哮喘

本文列举了人类存在Ｔ辅助细胞（Ｔ＿Ｈ）功能性亚群（Ｔ＿Ｈ和Ｔ＿Ｈ２）的证据；叙述调节Ｔ＿Ｈ细胞亚群分化的因素，尤其是Ｔ＿Ｈ１和Ｔ＿Ｈ２的相互制约作用；并讨论了Ｔ＿Ｈ２细胞在支气管哮喘慢性气管炎症形成中的作用，为防治哮喘提供新的思路。     

支气管哮喘患儿外周血B细胞和T细胞及其亚群的研究

目的 :探讨了支气管哮喘患儿外周血B细胞和T淋巴细胞及其亚群的变化。方法 :应用单克隆技术测定了 38例支气管哮喘患儿外周血B细胞和T淋巴细胞亚群的水平并以 30名正常健康人作比较。结果 :支气管哮喘患儿外周血B细胞数显著地高于正常人组 (P <0 .0 1 )CD3 、CD4、CD4/CD8显著地低于正常人 (P <0 .0 1 )。结论 :支气管哮喘是一种自身免疫调节异常的疾病     

支气管哮喘大鼠γδT细胞分布和凋亡的变化

探讨γδT细胞在支气管哮喘外周血和支气管肺泡灌洗液 (BALF)中的分布和凋亡状态。应用鸡卵清蛋白(OVA)致敏和刺激Wistar大鼠 (每组 10只 ) ,制作致敏大鼠哮喘模型 ,收集外周血单个核细胞 (PBMC)和BALF ,采用流式细胞术检测γδTCR+ T细胞百分率和CD2 8 γδTCR平均荧光密度比 ,并用免疫荧光法结合HE染色以及免疫组化法检测γδT细胞占淋巴细胞的百分率 ,用TENUL法检测淋巴细胞凋亡。哮喘组PBMC中γδT细胞占总T细胞或总淋巴细胞比例明显低于正常组 (P <0 0 5 ) ,CD2 8 γδTCR平均荧光密度比则无明显差别 ;BALF中γδT细胞占总T细胞或总淋巴细胞比例明显高于正常组 (P <0 0 1) ,CD2 8 γδTCR平均荧光密度比也有显著增加 (P <0 0 1) ;哮喘组PBMC和BALF中淋巴细胞凋亡指数明显低于正常对照组 (P <0 0 1)。提示γδT细胞亚群参与了哮喘的发病过程。     

支气管肺泡灌洗液细胞学诊断肺泡蛋白沉着症1例

患者女性 ,43岁 ,工人。因慢性咳嗽、咳痰、进行性呼吸困难 4年余 ,加重伴低热 1年入院。患者既往有高分子粘合剂粉尘接触史 11年 ,油漆接触史 7年。胸部Ｘ线示 :两肺下野可见肺纹理增粗并连成网状 ,右下外带呈毛玻璃状改变 ,心肋膈角锐利。胸部ＣＴ示两肺中下肺野可见分布不均匀的片状毛玻璃样病灶。肺功能检查 :提示肺通气功能中度障碍。病理检查　送检支气管灌洗液标本 ,标本底部肉眼可见米黄色淤泥样沉积物。涂片镜下所见 :在纤毛柱状上皮细胞、炎细胞及少许吞噬细胞背景中 ,可见均匀粉染无结构球形小体 ,直径 2 0～ 5 0 μｍ ,球形小体…     

哮喘儿童支气管肺泡灌洗液IL-17、 IL-8和VEGF水平测定及意义

目的: Th17细胞是以分泌IL-17为主的新型T细胞亚群,其在哮喘中的作用尚不十分清楚。本文通过观察哮喘急性发作儿童支气管肺泡灌洗液(BALF)IL-17、IL-8、血管内皮生长因子(VEGF)等的水平变化,探讨其在哮喘急性发作儿童气道炎症中的作用。方法: 我院2009年2月-2009年12月期间行纤维支气管镜检查患儿共88例,包括哮喘急性发作组(哮喘组,n=52)、非喘息组(肺炎组,n=25)及对照组(n=11),收集所有病例的BALF,进行细胞学分类,ELISA法测定BALF中细胞因子IL-17、IL-8、VEGF、IL-4、IFN-γ和IL- 4/IFN-γ水平。结果: 与对照组比较,哮喘组和肺炎组患儿的IL-17和IL-8水平均明显增高(均P<0.05);哮喘组患儿的IL-8水平较肺炎组低(P<0.05),而2组IL-17水平无显著差异(P>0.05);与肺炎组和对照组比较,哮喘组患儿VEGF水平明显增高(均P<0.01);肺炎组与对照组患儿VEGF水平无显著差异(P>0.05);3组患儿的IL-4,IFN-γ和IL-4/IFN-γ水平均无显著差异(均P>0.05)。与对照组比较,哮喘组及肺炎组患儿中性粒细胞百分比明显增高(均P<0.01),而哮喘组与肺炎组患儿的中性粒细胞百分比无显著差异(P> 0.05)。结论: IL-17、IL-8及VEGF在哮喘儿童气道炎症中发挥重要作用,Th17细胞可能参与儿童哮喘急性发作的发病机制。     

Assessment of local cellular immunity in lung cancer by bronchoalveolar lavage

Summary Small cell lung cancer (SCLC) is the most malignant of the pulmonary neoplasms and is associated with a poor local cellular immune response. 16 patients with non small cell lung cancer (NSCLC) and 11 patients with SCLC underwent bronchoalveolar lavage (BAL) in the lung which harbored the tumor in order to investigate the lymphocyte surface antigens utilizing the immunoperoxidase technique. Analysis of blood lymphocytes was performed in parallel. 8 patients with previous sarcoidosis in complete remission who underwent BAL and 10 normal blood donors served as controls.Among blood lymphocytes the CD3⁺, CD4⁺ and CD16⁺ cell populations were elevated significantly and the T4/T8 ratio was elevated in NSCLC patients, but only CD16⁺ were augmented in SCLC. Cell populations expressing the activation markers transferrin (TF) receptor, interleukin-2 (IL-2) receptor and the very late antigen VAL-1 were also increased in NSCLC, while SCLC was associated with antigen distributions similar to controls. No differences between the cohorts were seen in the expression of human leukocyte antigen (HLA)-DR. In BAL the population of CD3⁺ and CD4⁺ cells were reduced in SCLC and the T4/T8 ratio was diminished in contrast to controls and NSCLC patients, whereas these two latter groups did not differ from each other. The distribution pattern of CD16, TF receptor and IL-2 receptor in the study groups resembled that of cells of the blood stream, but CD16⁺ natural killer cells were additionally down regulated to control values in SCLC. No differences were seen in the distribution of VLA-1. HLA-DR⁺ cells were clearly elevated in both cancer groups.In general NSCLC was associated with a shift to higher relative numbers of immunocompetent and activated cells. This was most probably attributable to an immune response to neoplastic growth. This shift was largely lacking in SCLC. The analysis of lymphocytes from the periphery of the target organ emerged as a sensitive tool for the study of cellular immunity in lung cancer and showed many similarities to circulating blood cells. However, the analysis of natural killer cells and HLA-DR suggested a dissection of cellular immune response between blood and lung in pulmonary cancer. A depressive interaction between the tumor and the cellular host immune response may contribute to the exceptional malignancy of SCLC.Abbreviations BAL bronchoalveolar lavage - HLA-DR human leukocyte antigen-DR - IL-2 interleukin-2 - NSCLC non small cell lung cancer - SCLC small cell lung cancer - TF transferrin - VLA-1 very late antigen-1     

Effect of transportation stress on blood and bronchoalveolar lavage fluid components in calves

The effect of transportation stress on the content of bronchoalveolar lavage (BAL) fluid of 20 male Holstein–Friesian calves, 4–10 months old (mean weight 160 kg) was studied. The calves were healthy and had no previous history of respiratory tract diseases. During a period of 42 days experiment, the calves were kept indoors and were fed alfalfa hay and corn silage ad libitum. After a period of adaptation, on day 21, BAL fluid, blood samples, and nasal swabs were taken from all calves; then, the calves were divided into three groups: experimental (ten calves), which were transported and were deprived of food and water during transportation; control 1 (five calves), which were not transported and had free access to food and water during the 12 h of transportation of the experimental group; and control 2 (five calves), which were not transported but were deprived of food and water for the same time as the experimental group. On day 26, BAL fluid samples and nasal swabs were taken from control group 1. Blood samples were collected simultaneously from all groups at 0, 1, 3, 6, and 12 h of transportation. On days 27, 31, and 42, all previous samplings (BAL fluid, blood, and nasal swabs) were conducted on the experimental group and control group 2. Cytological, biochemical, and bacteriologic examination of BAL fluid and hematological and biochemical examination of blood samples revealed that the number of red blood cells, white blood cells, neutrophils, and the levels of cortisol, packed cell volume, total protein, and fibrinogen significantly increased, but lymphocytes significantly decreased in the experimental group compared with control groups 1 and 2 on the day of transportation (p < 0.05). In addition, regarding BAL fluid content, total cell count, macrophages, neutrophils, and total protein increased in the experimental group (p < 0.05). Pasteurella multocida was isolated from BAL fluid of three calves in the experimental group after transportation. Alteration in BAL fluid components in this study may be due to a depressed efficiency of mucociliary system and/or decreased amount of alveolar spatial surfactant either or both of which may predispose affected livestock to show the presence of P. multocida in bronchoalveolar fluid.     

Secretion of the eosinophil-active cytokines interleukin-5, granulocyte/macrophage colonystimulating factor and interleukin-3 by bronchoalveolar lavage CD4+ and CD8+ T cell lines in atopics asthmatics,and atopic and nonatopic controls

Specific eosinophil accumulation and activation within the asthmatic bronchial mucosa are thought to occur at least partly through the actions of cytokines, including interleukin (IL)-5, IL-3 and granulocyte/macrophage colonystimulating factor (GM-CSF). Although mRNA encoding some of these cytokines has been demonstrated in bronchoalveolar lavage (BAL) fluid cells and bronchial biopsies from asthmatics, it has yet to be established whether these cells produce the translated products and whether expression is associated with CD4⁺ T helper or CD8⁺ cytotoxic T cells. We addressed this problem by raising polyclonal CD4⁺ and CD8⁺ T cell lines from the BAL fluid of six atopic asthmatics, five atopic non-asthmatics and seven non-atopic non-asthmatic controls. BAL fluid cells obtained at fiberoptic bronchoscopy were depleted of adherent cells, and then T lymphocytes expanded by stimulation with monoclonal anti-CD3 antibody and recombinant human IL-2. When lymphocytes had expanded to sufficient numbers, CD4⁺ and CD8⁺ cells were separated by positive selection with magnetic beads coated with anti-CD4 or anti-CD8 monoclonal antibodies and further expanded. Cytokine secretion by standardized cell numbers was measured by enzyme-linked immunosorbent assays. BAL CD4⁺ T cell lines from the asthmatics secreted significantly elevated quantities of both IL-5 and GM-CSF as compared with lines from the atopic and non-atopic controls (p = 0.023–0.003). In contrast, IL-3 secretion did not significantly differ between the groups. In some subjects, CD8⁺ T cell lines also secreted significant quantities of these cytokines and there was a trend for IL-5 secretion by these cells to be higher in asthmatics than non-atopic controls (p = 0.035). These data are consistent with the hypothesis that activated T lymphocytes from asthmatics, particularly of the CD4⁺ subset, are predisposed to release elevated quantities of cytokines relevant to the accumulation and activation of eosinophils.     

高氧致早产鼠BALF及肺组织中MDA、SOD的变化

目的观察丙二醛(MDA)、超氧化物歧化酶(SOD)在高氧致(CLD)早产鼠支气管肺泡灌洗液(BALF)及肺组织中的变化.方法用高浓度氧致早产鼠CLD为研究对象,应用生化方法同步检测肺组织和BALF中MDA含量及SOD的活性.结果实验组肺组织中SOD的活性呈升高趋势,但与对照组比较,肺组织和BALF中SOD的活性均无差异(P>0.05、P>0.05),实验组肺组织和BALF中MDA的水平呈同相变化,自吸入高氧的第3d开始升高,(P<0.05),7d达高峰并持续至14d(P<0.01),21d时虽有下降,但仍高于对照组(P<0.05).结论 SOD、MDA的动态变化,可间接反应肺部疾病的变化,对慢性肺疾病的诊断、鉴别诊断、活动性判断及预后均有一定的价值.同时也证实机体的氧化与抗氧化失衡是高氧所致早产儿慢性肺疾病的发病原因之一.     

Foamy alveolar casts: diagnostic specificity for Pneumocystis carinii pneumonia in bronchoalveolar lavage fluid cytology

Pneumocystis carinii pneumonia (PCP) is a major infectious complication of immunodeficiency states, including the acquired immunodeficiency syndrome (AIDS). Bronchoalveolar lavage (BAL) is a safe and effective procedure for making this diagnosis. In addition to the characteristic organisms, both histologic and cytologic material often reveals exudate in the form of foamy alveolar casts (FACs). To test the diagnostic utility of FACs in BAL fluids, we compared 20 PCP-positive and 28 PCP-negative fluids as assessed by silver stains. All PCP-positive fluids contained FACs on Papanicolaou-stained material. Only one PCP-negative lavage contained FACs, and transbronchial biopsy in this case revealed PCP. We suggest that FACs in BAL fluids are highly sensitive and specific for the diagnosis of PCP.     

Evaluation and comparison of molecular and conventional diagnostic modalities for detecting pulmonary tuberculosis in bronchoalveolar lavage fluid

ContextIn cases of sputum smear-negative and sputum-scarce (SSN/SC) pulmonary tuberculosis (PTB), bronchoalveolar lavage (BAL) fluid may be helpful in establishing diagnosis. No specific recommendations for BAL samples have yet been formulated due to limited literature.Aims1. To find a sensitive and specific protocol for same-day diagnosis of PTB using BAL in SSN/SC clinically suspected patients. 2. To evaluate the need to routinely perform MGIT for all BAL samples.Settings and DesignProspective observational study design in a tertiary care hospital in New Delhi.Methods and materialFibreoptic bronchoscopy was performed and BAL collected from 175 clinically suspected SSN/SC PTB patients. BAL samples were subjected to: ZN Stain, Xpert MTB/RIF CBNAAT, BACTEC MGIT 960 liquid culture and M. tuberculosis complex DNA Real time PCR. The results of the various diagnostic tests were analysed using a) MGIT as gold standard and b) a composite reference standard (CRS) for a final diagnosis of PTB.Statistical analysis usedMicrosoft Excel 2016 and SPSS version 21.0 were used. Sensitivity, specificity and predictive values were calculated and compared using McNemar test. A p value of <0.05 was considered statistically significant.Results34 Cases had a final diagnosis of TB as per the CRS. Using CRS, MGIT had a sensitivity of 50.0% (32.4%–67.6%). There was no statistically significant difference between sensitivities of CBNAAT and PCR; both were more sensitive than ZN stain. Sensitivity and specificity of CBNAAT was 79.4% (62.1%–91.3%) and 100.0% (97.4%–100.0%) respectively. The preferred protocol for the hospital is CBNAAT and ZN stain. There was no statistically significant difference in sensitivity by adding PCR or MGIT to this protocol.ConclusionsWe found it a good strategy to perform CBNAAT and ZN stain on BAL fluid for accurate and same-day PTB diagnosis. CBNAAT is useful for ruling PTB in even when BAL cultures are negative. It is prudent to continue to routinely perform MGIT for all BAL samples.     

Elevated levels of soluble TNF receptors in bronchoalveolar lavage fluid in extrinsic allergic alveolitis

BACKGROUND: Soluble TNF receptors (sTNFR1 and sTNFR2) are inhibitors of TNF and can block TNF bioactivity. TNF plays an important role in the development of extrinsic allergic alveolitis (EAA). OBJECTIVE: To evaluate whether sTNFR1 and sTNFR2 are locally increased in EAA. METHODS: We measured sTNFR1 and sTNFR2 in bronchoalveolar lavage fluid (BALF) and serum from nine EAA patients and 11 control subjects using an ELISA method. RESULTS: BALF sTNFR1 and sTNFR2 levels were 0.24+/- 0.04 ng/mL and 0.59+/-0.16 ng/mL in EAA patients, and thus significantly elevated in comparison with the controls (0.13+/-0.02 ng/mL and 0.08+/-0.04 ng/mL, both P<0.05). Serum sTNFR levels were not significantly different between the two groups. Both sTNFR1 and sTNFR2 concentrations in BALF correlated significantly with the lymphocyte percentage of BALF (r = 0.57 and 0.81, respectively). CONCLUSION: The two alveolar sTNFRs, particularly sTNFR2, may be involved in the pathogenesis of EAA as counter-regulators of TNF.     

Low specificity of the bacterial index for the diagnosis of bacterial pneumonia by bronchoalveolar lavage

The bacterial index (BI) as defined by the sum of log₁₀ colony-forming units (cfu) of microorganisms per milliliter of bronchoalveolar lavage (BAL) fluid, i.e., a multiplication of the single cfu/ml, has been used to distinguish between polymicrobial pneumonia (BI5) and colonization (BI<5). Since many false-positive results are to be expected using this parameter, the diagnostic value of the BI was studied prospectively by obtaining bacteriologic cultures of BAL fluid in 165 consecutive unselected patients. In 27 cases the diagnosis of bacterial pneumonia was established on clinical criteria. In 133 patients pneumonia could be excluded, and in five patients the diagnosis remained unclear. Using a cut-off of 10⁵ cfu/ml BAL fluid, sensitivity and specificity for the diagnosis of pneumonia were 33% (9/27) and 99% (132/133), respectively. Sensitivity was mainly influenced by prior treatment with antibiotics, being 70% (7/10) in untreated and 12% (2/17) in treated patients. Applying the BI methodology at a cut-off of 5, however, resulted in an unacceptably high rate of 16 additional false-positive results, thus lowering the specificity to 87% (116/133;P<0.0001) while increasing the sensitivity to only 41% (11/27;P=0.77). In conclusion, given the high rate of false-positive results, the methodology of the BI is of doubtful value for the diagnosis of bacterial pneumonia by BAL in an unselected patient group. By applying the absolute number of cfu/ml BAL fluid, however, positive bacteriologic cultures of BAL fluid are highly specific for the diagnosis of pneumonia. Their sensitivity is limited by previous antibiotic therapy.     

IL-6和IL-8 mRNA在变态反应性哮喘患者BAL细胞的表达

目的：测定IL-6和IL-8 mRNA在变态反应性哮喘患者支气管肺泡灌洗（BAL）细胞中的表达。方法：采用原位杂交技术，测定变态反应性哮喘患者BAL中表达IL-6和IL-8 mRNA的阳性细胞数。结果：全部哮喘患者BAL细胞对IL-6和IL-8 mRNA探针杂交均呈阳性（阳性率分别为14/14和14/14）。变态反应性哮喘患者BAL中表达IL-6和IL-8 mRNA阳性细胞数明显多于对照（P<0.01）。结论：变态反应性哮喘患者BAL细胞中IL-6和IL-8 mRNA表达增加。