2009甲型H1N1流感患儿免疫功能改变初探
2009갑형H1N1류감환인면역공능개변초탐
Changes of immune function in patients with 2009 influenza A ( H1N1 )
  • 万方数据
    中华儿科杂志 중화인과잡지
    Chinese Journal of Pediatrics
    2010年 12期 947-953 ,共7页

    李成荣%杨军%贾实磊%王国兵%何颜霞%祖莹%杨卫国%付丹

    리성영%양군%가실뢰%왕국병%하안하%조형%양위국%부단

    流感,人%免疫活性%流感病毒A型,H1N1亚型 流感,人%免疫活性%流感病毒A型,H1N1亞型
    류감,인%면역활성%류감병독A형,H1N1아형
    Influenza,human%Immunocompetence%Influenza A virus,H1N1 subtype
    目的 探讨2009甲型H1N1流感(以下简称"甲流")患儿免疫功能及可能的免疫发病机制.方法 深圳市儿童医院2009年11月1日-2010年1月10日甲流住院患儿60例,轻症35例(轻症肺炎),重症25例(重症肺炎或甲流相关性脑病,死亡3例),同年龄正常对照组20例.采用real-time PCR、流式细胞术及ELISA检测外周血单个核细胞胞浆模式识别受体(PRRs)维甲酸诱导基因I/黑色素瘤分化相关基因5(RIG/MDA5)、胞膜PRRs Toll样受体(TLRs)分子及其信号途径传导分子、细胞因子/趋化因子及负性调节因子变化;T、B及NK细胞凋亡及凋亡相关基因TRAIL和CASPASE-3表达.结果 (1)甲流患儿RIG/MDA5表达、TLR2、TLR4表达明显高于正常对照组[TLR2(9.69±3.15)×10-2vs.(3.96±0.83)×10-2,t=10.16,P＜0.05;TLR4(10.23±2.85)×10-2vs.(7.46±2.18)×10-2,t=3.76,P＜0.05],以重症甲流患儿增高为著,RIG/MDA5表达增高最为明显;TLRs途径信号传导分子MyD88、TRAM等表达明显高于轻症甲流患儿.(2)甲流患儿CD3+[(1.22±0.38)×109/Lvs.(3.59±1.10)×109/L,t=9.21,P＜0.05]、CD4+、CD8+T细胞及NK细胞绝对计数明显低于正常对照组,B细胞无明显改变.(3)轻症甲流患儿TNF-α、IL-6、IL-1β等炎症细胞因子血浓度或基因高于正常对照组,重症患儿炎症细胞因子TNF-α[(6.42±1.76)×10-2vs.(9.05±2.51)×10-2,t=4.55,P＜0.05]明显低于正常对照组.IFN-α/β表达持续高于正常对照组,尤以重症甲流患儿为著;IFN-I诱导基因IP-10[(20.52±6.09)×10-2vs(1.18±0.34)×10-2,t=18.74,P＜0.05]、RANTES或iNOS轻症患儿表达高于正常对照组,重症患儿表达则趋于减少.(4)甲流患儿CD3+[(32.90±7.66)%vs.(20.21±6.58)%,t=6.21,P＜0.05]、CD4+、CD8+T细胞、NK细胞凋亡高于正常对照组,以重症患儿更为显著.凋亡相关基因TRAIL和CASPASE-3表达明显高于正常对照组.(5)重症患儿PRRs负性凋节因子SOCS1、SOCS3、IRAK-M、TRAF4及FLN29表达明显高于轻症患儿,抗炎细胞因子IL-10及IL-10/TNFα比值随病情加重增高.结论 甲流患儿机体免疫功能紊乱,轻症患儿处于全身免疫激活状态,重症患儿同时存在免疫激活/免疫抑制反应.
    목적 탐토2009갑형H1N1류감(이하간칭"갑류")환인면역공능급가능적면역발병궤제.방법 심수시인동의원2009년11월1일-2010년1월10일갑류주원환인60례,경증35례(경증폐염),중증25례(중증폐염혹갑류상관성뇌병,사망3례),동년령정상대조조20례.채용real-time PCR、류식세포술급ELISA검측외주혈단개핵세포포장모식식별수체(PRRs)유갑산유도기인I/흑색소류분화상관기인5(RIG/MDA5)、포막PRRs Toll양수체(TLRs)분자급기신호도경전도분자、세포인자/추화인자급부성조절인자변화;T、B급NK세포조망급조망상관기인TRAIL화CASPASE-3표체.결과 (1)갑류환인RIG/MDA5표체、TLR2、TLR4표체명현고우정상대조조[TLR2(9.69±3.15)×10-2vs.(3.96±0.83)×10-2,t=10.16,P＜0.05;TLR4(10.23±2.85)×10-2vs.(7.46±2.18)×10-2,t=3.76,P＜0.05],이중증갑류환인증고위저,RIG/MDA5표체증고최위명현;TLRs도경신호전도분자MyD88、TRAM등표체명현고우경증갑류환인.(2)갑류환인CD3+[(1.22±0.38)×109/Lvs.(3.59±1.10)×109/L,t=9.21,P＜0.05]、CD4+、CD8+T세포급NK세포절대계수명현저우정상대조조,B세포무명현개변.(3)경증갑류환인TNF-α、IL-6、IL-1β등염증세포인자혈농도혹기인고우정상대조조,중증환인염증세포인자TNF-α[(6.42±1.76)×10-2vs.(9.05±2.51)×10-2,t=4.55,P＜0.05]명현저우정상대조조.IFN-α/β표체지속고우정상대조조,우이중증갑류환인위저;IFN-I유도기인IP-10[(20.52±6.09)×10-2vs(1.18±0.34)×10-2,t=18.74,P＜0.05]、RANTES혹iNOS경증환인표체고우정상대조조,중증환인표체칙추우감소.(4)갑류환인CD3+[(32.90±7.66)%vs.(20.21±6.58)%,t=6.21,P＜0.05]、CD4+、CD8+T세포、NK세포조망고우정상대조조,이중증환인경위현저.조망상관기인TRAIL화CASPASE-3표체명현고우정상대조조.(5)중증환인PRRs부성조절인자SOCS1、SOCS3、IRAK-M、TRAF4급FLN29표체명현고우경증환인,항염세포인자IL-10급IL-10/TNFα비치수병정가중증고.결론 갑류환인궤체면역공능문란,경증환인처우전신면역격활상태,중증환인동시존재면역격활/면역억제반응.
    Objective To investigate the alteration of immune function and possible immunopathogenesis in the children with 2009 influenza A ( H1N1 ) infection. Method Sixty patients with 2009 influenza A ( H1 N1 ) infection hospitalized in Shenzhen Children's Hospital between November 1,2009 and January 10, 2010 and 20 age-matched healthy children were enrolled in this study. The patients were divided into two groups according to the severity of influenza A infection: 35 mild cases (mild pneumonia)and 25 severe cases ( severe pneumonia, acute encephalopathy associated with influenza A, and 3 died from acute necrotizing encephalopathy with influenza A infection). Real-time PCR was used to evaluate the expression levels of pattern recognition receptor (PRRs), retinoic acid induced gene I/melanoma differentiation associated gene 5 ( RIG/MDA5 ), Toll-like receptors (TLRs) and TLRs signaling molecules,and negative-regulator. Three color fluorescent and flow cytometry were used to investigate the apoptosis of CD3+, CD4+, CD8+ and CD19+ cells. Plasma cytokines (IL-1β, IL-6, TNF-α, IFN-γ, IFN-α, IL-10)concentrations were measured by enzyme-linked immunosorbent assay (ELISA). Result (1) The expression levels of RIG/MDA5, TLR2, 4 were much higher in the patients with influenza A infection,especially severe cases [ TLR2 (9. 69 ± 3. 15 ) × 10 - 2 vs. (3.96 ± 0. 83 ) × 10 - 2, t = 10. 16,P ＜ 0. 05; TLR4 ( 10. 23 ± 2. 85 ) × 10-2 vs. (7. 46 ± 2. 18 ) × 10-2, t = 3.76, P ＜ 0. 05 ]. The expression levels of TLRs signal transduction molecules like MyD88 and TRAM also increased. (2) The cell counts of CD3 + , CD4 + ,CD8 + T cells and NK cells were markedly lower in the patients with influenza A infection compared to the NC group [CD3 + (1.22 ±0. 38) × 109/L vs. (3.59 ± 1.10) × 109/L,t =9.21 ,P ＜0. 05]. (3) Plasma concentrations and the mRNA expression of TNF-α, IL-6, and IL-1β were elevated in mild cases, while declined in severe cases [ TNF-α ( 6. 42 ± 1.76 ) × 10 -2 vs. ( 9.05 ± 2. 51 ) × 10 - 2, t = 4. 55, P ＜ 0. 05 ].Plasma concentrations of IFN-α/IFN-β were up-regulated gradually with the aggravation of the disease,especially in severe cases. Compared with healthy controls, the expression of IFN-I inducible gene 1P-l0,RANTES, or iNOS was significantly higher in children with mild [ IP-10(20. 52 ±6. 09) × 10-2 vs. ( 1.18 ±0. 34) × 10 -2 ,t = 18. 74, P ＜ 0. 05], and relatively lower in severe cases. (4) The apoptosis of CD3 +,CD4 +, CD8 + and NK cells significantly increased in the patients with influenza A infection than those in NC group [CD3+ (32.90 ±7.66)% vs. (20.21 ±6.58)%,t =6.21,P＜0.05]. Compared with healthy controls, the expression levels of apoptosis-related gene like TRAIL and CASPASE-3 significantly increased in the patients with influenza A infection. (5)The expression levels of negative regulator of SOCS1, SOCS3,IRAK-M, TRAF4 and FLN29 were significantly increased in the patients with influenza A, especially in severe cases than those in NC group (P ＜ 0. 05). Conclusion Immune function changed with the severity of the disease. The mild cases presented systemic immune activation status, while critically ill cases presented mixed immune activation and immunosuppression status.
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