摘要
目的了解人类博卡病毒在小儿呼吸道感染时其急性期外周血细胞因子的变化,旨在探讨该病毒感染的免疫致病机制。
材料和方法
材料:收集2010年3月～2010年9月广东省汕头大学医学院第二附属医院儿科因急性呼吸道感染住院患儿的咽拭子及其对应外周血血清574份,同时采集92例健康体检儿童呼吸道的咽拭子及其对应外周血血清作为对照研究。
方法:1、应用PCR技术进行人类博卡病毒筛查,同时通过PCR或RT-PCR技术对上述咽拭子进行其他9种病毒(呼吸道合胞病毒、WU多瘤病毒、人偏肺病毒、腺病毒、流感病毒A,B型、副流感病毒1,3型和鼻病毒)筛查。2、在上述方法中筛查出的单纯人类博卡病毒阳性标本和单纯呼吸道合胞病毒阳性标本中找出对应的外周血血清标本,随机从中选出29份人类博卡病毒感染组血清和26例呼吸道合胞病毒感染组血清,同时随机挑选30例正常对照组血清。通过Elisa原理,应用液相蛋白芯片(Luminex)技术对3组血清中12种细胞因子(IL-1α、IL-1β、IL-2、IL-4、IL-6、IL-8、IL-10、IL-12p40、IL-12p70、IL-13、IFN-γ、TNF-α)的含量进行检测,并对相关检测结果进行统计和分析。
结果
1、病毒筛查结果:
574例患儿人类博卡病毒阳性92例(占16.02%),其中单纯人类博卡感染43例(46.73%),合并其他病毒感染的有49例(53.27%),分别为呼吸道合胞病毒(17例)、流感病毒A型(14例)、副流感病毒3型(10例)、副流感病毒1型(8例)、腺病毒(8例)、WU多瘤病毒(7例)、鼻病毒(1例)。呼吸道合胞病毒阳性68例(占11.84%),其中单纯呼吸道合胞病毒感染28例(41.18%),合并感染40例(58.82%),分别为流感病毒A型(22例)、人类博卡病毒(17例)、副流感病毒3型(13例)、WU多瘤病毒(10例)、腺病毒(7例)、副流感病毒1型(6例)、鼻病毒(1例)。92例健康体检儿童10种病毒的PCR筛查均阴性。
2、外周血细胞因子检测结果:
①单纯人类博卡病毒感染组患儿外周血IL-6、IL-8含量分别为27.71(136.78)、18.75(216.08),均较正常对照组明显升高,差异有统计学意义(P均<0.05)。
②单纯呼吸道合胞病毒感染组患儿外周血IL-1β、IL-2、IL-4、IL-6、IL-8、IL-10、IL-12p40、IL-13、TNF-α的含量分别为3.21(3.00)、4.00(4.16)、2.00(1.78)、398.5(984.23)、246.00 (663.00)、3.00(2.78)、242.50(483.28)、3.00(1.68)、3.00(1.24),均较正常对照组明显升高,差异有统计学意义(P均<0.05)。
③单纯人类博卡病毒感染组IL-1β、IL-6、IL-8、IL-12p40、IL-12p70、TNF-α含量分别为1.84 (1.47)、27.71(136.78)、18.75(216.08)、41.00(114.65)、8.11(5.77)、2.76(0.42),单纯呼吸道合胞病毒感染组IL-1β、IL-6、IL-8、IL-12p40、IL-12p70、TNF-α的含量分别为3.21(3.00)、398.5(984.23)、246.00 (663.00)、242.50(483.28)、4.00(4.56)、3.00(1.24),两组这6中细胞因子含量均较正常对照组升高,且两组间差异均有统计学意义(P均<0.05)。
④单纯人类博卡病毒感染组IL-1α、IL-2、IL-4、IL-10、INF-γ的含量分别为1.22 (6.28)、1.22 (0.78)、1.22 (0.78)、1.22 (0.78)、1.22 (4.28),INF-γ/ IL-4比值为1,均比正常对照组降低,但与正常对照组差异均无统计学意义(P>0.05);而RSV组这5种细胞因子含量分别为9.00(9.78)、4.00(4.16)、2.00(1.78)、3.00(2.78)、6.00(4.78),INF-γ/ IL-4比值为3,均比单纯人类博卡病毒感染组高,其中IL-1α、IL-2、IL-10水平两组差异有统计学意义(P均<0.05),两组IL-4、INF-γ水平差异无统计学意义(P>0.05)。
结论人类博卡病毒感染同呼吸道合胞病毒感染一样可导致患儿相应的免疫功能紊乱,且其免疫变化同普通呼吸道合胞病毒感染引起的免疫变化相似,程度相对较轻。机体抗人类博卡病毒感染免疫过程中存在细胞Th1/Th2失衡和Th2类细胞因子占优势。
Objective To investigate the immune pathological mechanism of Human Bocavirus (HBoV) which is in acute respiratory tract infection in children through the changes of the level of cytokines in peripheral blood.
Material and method
Material: From March to September 2010,throat swab samples and serum were collected from 574 children who were hospitalized with acute respiratory tract infection in Second Affiliated Hospital of Shantou University Medical College, and 92 were asymptomatic who visited the health checkup clinic.
Method: 1、HBoV was detected by using PCR technology, meanwhile 9 commonly respiratory infectious viruses (respiratory syncytial virus; WU polyomavirus; human metapneumovirus; adenovirus ; in?uenza A and B; parainfluenza 1 and 3; rhinovirus) were detected by using PCR or RT-PCR..2、The cytokines (IL-1α、IL-1β、IL-2、IL-4、IL-6、IL-8、IL-10、IL-12p40、IL-12p70、IL-13、IFN-γ、TNF-α) in serum of peripheral blood, in which collected from 30 healthy children,29 patients solely infected with HBoV and 26 patients solely infected with RSV were detected by using Luminxe simultaneously.
Result
1. Virus screening results:
In this study, 92 of the 574 tested specimens with acute respiratory tract infection were positive for HBoV (16.02%). HBoV was the sole virus detected in 43 specimens (46.73%) from patients with acute respiratory tract infection, and 49 of 92 (53.27%) specimens infected with HBoV were co-infected with other viruses, such as RSV, IVA、ADV、PIV1、PIV3. Among the 68 tested specimens with acute respiratory tract infection which were positive for RSV, only 28 were infected solely and 40 were co-infected. Amony the tested specimens, they were 22 of IVA, 17 of HBoV 13 of PIV3, 10 of WUPyV, 7 of ADV, 7 of PIV1 and 1 of Rhv. Among 92 specimens of children's health checkup, the PCR of ten viruses which was checked was negative.
2. Peripheral cytokine detection results:
①The level of peripheral blood IL-6 and IL-8 is 27.71(136.78)、18.75(216.08),which were the HBoV group was higher than the control group. The difference has statistical significance (P<0.05).
②Levels of IL-1β、IL-2、IL-4、IL-6、IL-8、IL-10、IL-12p40、IL-13、TNF-αin RSV group are 3.21(3.00)、4.00(4.16)、2.00(1.78)、398.5(984.23)、246.00 (663.00)、3.00(2.78)、242.50(483.28)、3.00(1.68)、3.00(1.24),which were the RSV group was higher than the control group,The difference has statistical significance (P<0.05)..
③Levels of IL-1β、IL-6、IL-8、IL-12p40、IL-12p70、TNF-αare1.84 (1.47)、27.71(136.78)、18.75(216.08)、41.00(114.65)、8.11(5.77)、2.76(0.42) in the HBoV group, and levels of IL-1β、IL-6、IL-8、IL-12p40、IL-12p70、TNF-α3.21(3.00)、398.5(984.23)、246.00 (663.00)、242.50(483.28)、4.00(4.56)、3.00(1.24),both of two groups are higher than control group,and there are statistical significance (P<0.05).
④Levels of IL-1α、IL-2、IL-4、IL-10、INF-γin the HBoV group are 1.22 (6.28)、1.22 (0.78)、1.22 (0.78)、1.22 (0.78)、1.22 (4.28),INF-γ/ IL-4=1,all are lower than the control group, but there is no statistical significanc(P>0.05).The levels of the five cytokine in the RSV group are 9.00(9.78)、4.00(4.16)、2.00(1.78)、3.00(2.78)、6.00(4.78),INF-γ/ IL-4比=3, All higner than HBoV group, The levels of IL-1α、IL-2、IL-10 between the HBoV group and the RSV group all are statistical significanc, but thsre is no statistical significanc of IL-4、INF-γ.
Conclusion Infection with HBoV could made abnormal immune function in children with acute respiratory tract infection, and the pattern was similar to those children infected with RSV,but the degree of the changes of cytokines was lower than those infected with RSV. The Th2 cytokines may be the Main role and Th1/Th2 is imbalances in the Infection with HBoV.
引文
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