流式细胞磁珠微阵列法检测血清中细胞因子水平在稳定期和急性加重期哮喘中的意义
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摘要
目的探讨流式细胞磁珠微阵列法(cytometric beads array,CBA)检测血清中细胞因子水平在稳定期和急性加重期哮喘中的意义。方法纳入慢性持续期哮喘40例,急性发作期哮喘22例,健康人18例。抽取外周血2 mL,采用CAB法检测外周血清中细胞因子水平,并分析各细胞因子特征及与临床数据的相关性。全自动血细胞及生化仪检测血常规,C反应蛋白和降钙素原。结果定制含7种细胞因子微球(分别为IL-5,IL-6,IL-10,IL-13,IL-17,TNF-α,IFN-γ)的CBA试剂盒。与健康人相比,7种细胞因子在慢性持续期哮喘无明显改变。急性发作期哮喘细胞因子升高分为5种亚群,分别为IL-6升高型(22.8%)、IL-6/TNF-α双高型(13.6%)、IL-5升高型(31.8%)、IL-17升高型(占13.6%)及细胞因子正常型(占18.2%)。其中,IL-6、TNF-α明显升高,较健康人分别升高了6.28~12倍和17.3倍。IL-6升高型、IL-6/TNF-α升高型与白细胞及C反应蛋白呈正相关(R2=0.63,R2=0.67;P<0.05)。结论 CBA法可快速检测哮喘血清标本的多种细胞因子水平,细胞因子在慢性持续期无明显升高,但IL-6、TNF-α、WBC和CRP的综合指标有助于判断由细菌感染诱发的急性加重,并快速指导抗生素的使用。
Objective To detect cytokines in serum for chronic asthma and acute exacerbation by cytometric beads array method(CBA),and to analyze characteristics of cytokine subgroup and their relation to clinical data. Methods Fourty chronic asthma,22 acute exacerbation of asthma and 18 healthy subjects were included.A little amount of blood was drawn from each subject(2 mL)and used for measuring cytokines by CBA to analyze their characteristics and correlation with clinical data.Regular blood test,C reaction protein and procalcitonin were measured by full automatic blood cell and biochemical analyzer. Results CBA kit of cytokine panels were custom-made,which included IL-5,IL-6,IL-10,IL-13,IL-17,TNF-αand IFN-γ.No differences were found in cytokine levels in chronic asthma when compared with healthy subjects.However,in acute exacerbation phase,there were 5 cytokine subgroups which were high IL-6(22.8%),high IL-6/TNF-α(13.6%),high IL-5(31.8%),high IL-17(13.6%)and normal cytokine(18.2%).The levels of IL-6 and TNF-αincreased 6.28-12 folds and 17.3 folds respectively compared with healthy subiects.High IL-6 and High IL-6/TNF-αwere positively correlated with WBC and CRP(R2=0.63,R2=0.67:P<0.05). Conclusion CBA was a reliable way to detect muti-cytokines in asthma serum in a short time.No differences were found in cytokine levels in chronic asthma.However,the specific cytokine panel(IL-6,TNF-α,WBC and CRP)may be helpful to identify the acute exacerbation caused by bacterial and guide appropriate antibiotic therapy.
Objective To detect cytokines in serum for chronic asthma and acute exacerbation by cytometric beads array method(CBA),and to analyze characteristics of cytokine subgroup and their relation to clinical data. Methods Fourty chronic asthma,22 acute exacerbation of asthma and 18 healthy subjects were included.A little amount of blood was drawn from each subject(2 mL)and used for measuring cytokines by CBA to analyze their characteristics and correlation with clinical data.Regular blood test,C reaction protein and procalcitonin were measured by full automatic blood cell and biochemical analyzer. Results CBA kit of cytokine panels were custom-made,which included IL-5,IL-6,IL-10,IL-13,IL-17,TNF-αand IFN-γ.No differences were found in cytokine levels in chronic asthma when compared with healthy subjects.However,in acute exacerbation phase,there were 5 cytokine subgroups which were high IL-6(22.8%),high IL-6/TNF-α(13.6%),high IL-5(31.8%),high IL-17(13.6%)and normal cytokine(18.2%).The levels of IL-6 and TNF-αincreased 6.28-12 folds and 17.3 folds respectively compared with healthy subiects.High IL-6 and High IL-6/TNF-αwere positively correlated with WBC and CRP(R2=0.63,R2=0.67:P<0.05). Conclusion CBA was a reliable way to detect muti-cytokines in asthma serum in a short time.No differences were found in cytokine levels in chronic asthma.However,the specific cytokine panel(IL-6,TNF-α,WBC and CRP)may be helpful to identify the acute exacerbation caused by bacterial and guide appropriate antibiotic therapy.
引文
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[15]YIMIN KM,OZAKI M,HAGA S,et al.Mutual modulation between interleukin-10 and interleukin-6induced by Rhodococcus aurantiacus infection in mice[J].Microbes Infect,2008,10(14-15):1450-1458.
[16]LEE SL,CHIU SSS,MALIK PJ,et al.Is respiratory viral infection really an important trigger of asthma exacerbations in children?[J].Eur J Pediatr,2011,170(10):1317-1324.
[17]何剑,朱柠,陈小东.阿奇霉素对支气管哮喘患者肺功能及细胞因子的影响[J].复旦学报(医学版),2009,36(6):719-722.
[2]ISGRO M,BIANCHETTI L,MARINI MA,et al.Involvement of fibrocytes in allergen-induced T cell responses and rhinovirus infections in asthma[J].Biochem Biophys Res Commun,2013,437(3):446-451.
[3]LEONARD C,TORMEY V,BURKE C,et al.Allergeninduced cytokine production in atopic disease and its relationship to disease severity[J].Am J Respir Cell Mol Biol,1997,17(3):368-375.
[4]RUDIN A,MACAUBAS C,WEE C,et al.“Bystander”amplification of PBMC cytokine responses to seasonal allergen in polysensitized atopic children[J].Allergy,2001,56(11):1042-1048.
[5]SIMMS E,KJARSGAARD M,DENIS S,et al.Cytokine responses of peripheral blood mononuclear cells to allergen do not identify asthma or asthma phenotypes[J].Clin Exp Allergy,2013,43(11):1226-1235.
[6]饶锦秀,汪宏良,胡芳,等.哮喘患者血清IL-4与IFN-7水平的测定和临床意义[J].国际检验学杂志,2010,31(2):172-173.
[7]付海卫,张松,张添威.黄昌河布地奈德雾化吸入对哮喘急性发作期患者细胞因子和肺功能的影响[J].中国药业,2013,22(15):79-80.
[8]熊曙光,聂晓红,王慧,等.检测血清白细胞介素4、白细胞介素16和内皮素1在哮喘患者中的意义[J].中国呼吸与危重监护杂志,2008,7(5):385-387.
[9]HERBERTH G,DAEGELMANN C,WEBER A,et al.Association of neuropeptides with Th1/Th2 balance and allergic sensitization in children[J].Clin Exp Allergy,2006,36(11):1408-1416.
[10]KUWANO Y,FUJIMOTO M,WATANABE R,et al.Serum chemokine profiles in patients with alopecia areata[J].Br J Dermatol,2007,157(3):466-473.
[11]NADI E,ARJIPOUR M,SHARIFI S,et al.Assay of IL-22 and IL-25 in serum,whole blood,and peripheral blood mononuclear cell cultures of patients with severe asthma[J].Allergol Immunopathol(Madr),2014,42(5):402-406.
[12]MA Y,LIU X,WEI Z,et al.The expression and significance of TIPE2 in peripheral blood mononuclear cells from asthmatic children[J].Scand J Immunol,2013,78(6):523-528.
[13]BALAJI R,WRIGHT KJ,TURNER JL,et al.Circulating cortisol,tumor necrosis factor-alpha interleukin-1beta,and interferon-gamma in pigs infected with Actinobacillus pleuropneumoniae[J].J Anim Sci,2002,80(1):202-207.
[14]THANAWONGNUWECH R,THACKER B,HALBUR P,et al.Increased production of proinflammatory cytokines following infection with porcine reproductive and respiratory syndrome virus and Mycoplasma hyopneumoniae[J].Clin Diagn Lab Immunol,2004,11(5):901-908.
[15]YIMIN KM,OZAKI M,HAGA S,et al.Mutual modulation between interleukin-10 and interleukin-6induced by Rhodococcus aurantiacus infection in mice[J].Microbes Infect,2008,10(14-15):1450-1458.
[16]LEE SL,CHIU SSS,MALIK PJ,et al.Is respiratory viral infection really an important trigger of asthma exacerbations in children?[J].Eur J Pediatr,2011,170(10):1317-1324.
[17]何剑,朱柠,陈小东.阿奇霉素对支气管哮喘患者肺功能及细胞因子的影响[J].复旦学报(医学版),2009,36(6):719-722.