基于痰液粒细胞对患者分层后分析哮喘严重程度表型和炎症反应蛋白
2010/08/11
背景：与轻度和中度哮喘患者相比，重度哮喘患者痰液中粒细胞数目增加。
目的：我们认为，痰液炎症反应粒细胞有助于鉴别哮喘严重程度表型，而且与痰液上清液炎症反应蛋白相关。
方法：对重症哮喘研究计划（Severe Asthma Research Program）中的242名哮喘患者进行了研究。对患者痰液样本进行细胞计数、细胞组分检测，细胞裂解液Western blot 分析，ELISA和蛋白芯片分析痰液上清液炎症反应蛋白。
结果：根据痰液粒细胞对患者进行分层：<2%或≥2%的嗜酸性粒细胞和<40%或≥40%的中性粒细胞。中性粒细胞和嗜酸性粒细胞同时增加的患者肺功能最低，症状发生率增加，对护理的需求也增加。对于嗜酸性粒细胞增加，伴或不伴中性粒细胞增加的患者，呼出气一氧化氮分数（FeNO）、血清嗜酸性粒细胞和每日β受体激动剂的使用频率增加。以粒细胞分层后，蛋白芯片分析显示，对于痰液中性粒细胞≥40%的患者，痰液25～28种炎症反应蛋白增加大于2倍。以哮喘严重程度分层后，蛋白芯片显示，与非严重哮喘患者相比，严重哮喘患者痰液有6～9中蛋白增加大于2倍。痰液粒细胞分层后，ELISA 显示，中性粒细胞≥40%的患者，其脑源性神经营养因子、IL-1β和巨噬细胞炎症反应蛋白3α/CCL20表达增加，这些介质与中性粒细胞计数相关。
结论：痰液嗜酸性粒细胞和中性粒细胞同时增加的哮喘患者肺功能最差、哮喘控制较差、哮喘症状发生频率增加以及护理需求也增加。痰液上清液的炎症反应蛋白分析显示哮喘患者存在新的炎症介质表达增加，且与痰液中性粒细胞增加相关。
（林江涛 审校）
J Allergy Clin Immunol. 2010 May;125(5):1028-1036.e13. Epub 2010 Apr 15.
Analyses of asthma severity phenotypes and inflammatory proteins in subjects stratified by sputum granulocytes.
Hastie AT, Moore WC, Meyers DA, Vestal PL, Li H, Peters SP, Bleecker ER; National Heart, Lung, and Blood Institute Severe Asthma Research Program.
Center for Human Genomics, Wake Forest University Health Sciences, Winston Salem, NC 27157, USA. ahastie@wfubmc.edu
Abstract
BACKGROUND: Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma.
OBJECTIVE: We hypothesized that inflammatory granulocytes in sputum may identify specific asthma severity phenotypes and are associated with different patterns of inflammatory proteins in sputum supernatants.
METHODS: This hypothesis was tested in 242 patients with asthma enrolled in the Severe Asthma Research Program who provided sputum samples for cell count, differential cell determinations, cell lysates for Western blot, and supernatant analyses by inflammatory protein microarrays and ELISAs. ANOVA and multiple linear regression models tested mediator associations.
RESULTS: Stratified by sputum granulocytes, <2% or > or = 2% eosinophils and <40% or > or = 40% neutrophils, subjects with both increased eosinophils and neutrophils had the lowest lung function and increased symptoms and health care use. Subjects with elevated eosinophils with or without increased neutrophils had significantly increased fraction exhaled nitric oxide (FeNO) and serum eosinophils and greater frequency of daily beta-agonist use. Microarray data stratified by granulocytes revealed 25 to 28 inflammatory proteins increased >2-fold in sputa with > or = 40% neutrophils. Microarray analyses stratified by severity of asthma identified 6 to 9 proteins increased >2-fold in sputa in subjects with severe asthma compared with nonsevere asthma. ELISA data stratified by sputum granulocytes showed significant increases in brain-derived neurotrophic factor, IL-1beta, and macrophage inflammatory protein 3alpha/CCL20 for those with > or = 40% neutrophils; these mediators demonstrated positive associations with neutrophil counts.
CONCLUSION: Combined increased sputum eosinophils and neutrophils identified patients with asthma with the lowest lung function, worse asthma control, and increased symptoms and health care requirements. Inflammatory protein analyses of sputum supernatants found novel mediators increased in patients with asthma, predominantly associated with increased sputum neutrophils.
J Allergy Clin Immunol. 2010 May;125(5):1028-1036.e13. Epub 2010 Apr 15.
目的：我们认为，痰液炎症反应粒细胞有助于鉴别哮喘严重程度表型，而且与痰液上清液炎症反应蛋白相关。
方法：对重症哮喘研究计划（Severe Asthma Research Program）中的242名哮喘患者进行了研究。对患者痰液样本进行细胞计数、细胞组分检测，细胞裂解液Western blot 分析，ELISA和蛋白芯片分析痰液上清液炎症反应蛋白。
结果：根据痰液粒细胞对患者进行分层：<2%或≥2%的嗜酸性粒细胞和<40%或≥40%的中性粒细胞。中性粒细胞和嗜酸性粒细胞同时增加的患者肺功能最低，症状发生率增加，对护理的需求也增加。对于嗜酸性粒细胞增加，伴或不伴中性粒细胞增加的患者，呼出气一氧化氮分数（FeNO）、血清嗜酸性粒细胞和每日β受体激动剂的使用频率增加。以粒细胞分层后，蛋白芯片分析显示，对于痰液中性粒细胞≥40%的患者，痰液25～28种炎症反应蛋白增加大于2倍。以哮喘严重程度分层后，蛋白芯片显示，与非严重哮喘患者相比，严重哮喘患者痰液有6～9中蛋白增加大于2倍。痰液粒细胞分层后，ELISA 显示，中性粒细胞≥40%的患者，其脑源性神经营养因子、IL-1β和巨噬细胞炎症反应蛋白3α/CCL20表达增加，这些介质与中性粒细胞计数相关。
结论：痰液嗜酸性粒细胞和中性粒细胞同时增加的哮喘患者肺功能最差、哮喘控制较差、哮喘症状发生频率增加以及护理需求也增加。痰液上清液的炎症反应蛋白分析显示哮喘患者存在新的炎症介质表达增加，且与痰液中性粒细胞增加相关。
（林江涛 审校）
J Allergy Clin Immunol. 2010 May;125(5):1028-1036.e13. Epub 2010 Apr 15.
Analyses of asthma severity phenotypes and inflammatory proteins in subjects stratified by sputum granulocytes.
Hastie AT, Moore WC, Meyers DA, Vestal PL, Li H, Peters SP, Bleecker ER; National Heart, Lung, and Blood Institute Severe Asthma Research Program.
Center for Human Genomics, Wake Forest University Health Sciences, Winston Salem, NC 27157, USA. ahastie@wfubmc.edu
Abstract
BACKGROUND: Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma.
OBJECTIVE: We hypothesized that inflammatory granulocytes in sputum may identify specific asthma severity phenotypes and are associated with different patterns of inflammatory proteins in sputum supernatants.
METHODS: This hypothesis was tested in 242 patients with asthma enrolled in the Severe Asthma Research Program who provided sputum samples for cell count, differential cell determinations, cell lysates for Western blot, and supernatant analyses by inflammatory protein microarrays and ELISAs. ANOVA and multiple linear regression models tested mediator associations.
RESULTS: Stratified by sputum granulocytes, <2% or > or = 2% eosinophils and <40% or > or = 40% neutrophils, subjects with both increased eosinophils and neutrophils had the lowest lung function and increased symptoms and health care use. Subjects with elevated eosinophils with or without increased neutrophils had significantly increased fraction exhaled nitric oxide (FeNO) and serum eosinophils and greater frequency of daily beta-agonist use. Microarray data stratified by granulocytes revealed 25 to 28 inflammatory proteins increased >2-fold in sputa with > or = 40% neutrophils. Microarray analyses stratified by severity of asthma identified 6 to 9 proteins increased >2-fold in sputa in subjects with severe asthma compared with nonsevere asthma. ELISA data stratified by sputum granulocytes showed significant increases in brain-derived neurotrophic factor, IL-1beta, and macrophage inflammatory protein 3alpha/CCL20 for those with > or = 40% neutrophils; these mediators demonstrated positive associations with neutrophil counts.
CONCLUSION: Combined increased sputum eosinophils and neutrophils identified patients with asthma with the lowest lung function, worse asthma control, and increased symptoms and health care requirements. Inflammatory protein analyses of sputum supernatants found novel mediators increased in patients with asthma, predominantly associated with increased sputum neutrophils.
J Allergy Clin Immunol. 2010 May;125(5):1028-1036.e13. Epub 2010 Apr 15.
