【作者】 刘明慧；

【导师】 王素萍；

【作者基本信息】 山西医科大学 ， 流行病与卫生统计学， 2012， 硕士

【摘要】 目的①了解HBsAg阳性孕妇婴儿脐静脉血PBMC中TLR3与HBV宫内感染的关系。②探讨HBsAg阳性孕妇婴儿脐静脉血PBMC中相关免疫细胞与HBV宫内感染的关系。③探讨TLR3在HBsAg阳性孕妇婴儿HBV宫内感染中可能的作用机制。方法①收集2011年7月至2012年1月于太原市传染病医院妇产科分娩的HBsAg阳性孕妇产后脐血共52例。无菌采集分娩时脐血抗凝血30ml，并收集母儿的流行病学资料。②婴儿脐静脉血PBMC的分离及培养，体外HBV干预宫内感染组和非宫内感染组，以HBV+PolyI:C干预宫内感染组婴儿PBMC。③多色流式细胞术（FCM）对两组TLR3蛋白表达的荧光强度值以及T细胞、B细胞、DC细胞和CD4+CD25+Treg的百分比进行测定。结果①采用多色FCM测定TLR3蛋白表达荧光强度值，经比较：非宫内感染组的TLR3蛋白表达的平均荧光强度值（32.18±11.36）高于宫内感染组（30.29±11.04），差异无统计学意义（t=0.410,P=0.684）；宫内感染组TLR3活化后（即PolyI:C刺激后），TLR3蛋白表达的平均荧光强度值（32.16±9.40）升高，与非宫内感染组的（32.18±11.36）相比，无统计学差异（t=-1.824,P=0.078）。②采用多色FCM测定B细胞的百分比，经比较：非宫内感染组的B细胞的百分比（7.38±6.40）高于宫内感染组（6.76±4.01），没有统计学差异（t=0.210,P=0.834）；宫内感染组TLR3活化后，B细胞的百分比（5.14±2.64）降低，与非宫内感染组（7.38±6.40）比较，差异没有统计学意义（t=0.768,P=0.447）。③非宫内感染组的CD4+T细胞的百分比（41.14±20.08）高于宫内感染组（33.82±2.60），差异尚不能认为有统计学意义（t=0.618,P=0.543）；非宫内感染组的CD8+T细胞的百分比（11.30±5.87）与宫内感染组（11.14±9.64）相比，差异无统计学意义（t=0.041,P=0.968）；非宫内感染组的CD4+/CD8+比值（4.30±2.28）与宫内感染组的（4.75±3.23）比较，差异尚不能认为有统计学意义（t=-0.30,P=0.767）。④非宫内感染组的mDC的百分比（0.84±0.58）高于宫内感染组（0.67±0.34），差异尚不能认为有统计学意义（t=-0.485,P=0.633）；非宫内感染组的pDC（0.92±0.91）与宫内感染组的（0.91±0.46）比较，差异没有统计学意义（t=0.020,P=0.984）。⑤非宫内感染组的CD4~+CD25~+Treg细胞的百分比（4.69±3.41）与宫内感染组（3.00）比较，差异无统计学意义（t=0.474,P=0.646）。结论①非宫内感染组的TLR3蛋白表达平均荧光强度值高于宫内感染组，而宫内感染组经PolyI:C刺激后其表达水平升高，提示宫内感染可能与婴儿TLR3表达有关。②非宫内感染组的B细胞、CD4~+T细胞、mDC的百分比高于宫内感染组，提示宫内感染与婴儿B细胞、CD4~+T细胞、mDC表达可能有关。③本次研究未发现宫内感染与CD8~+T细胞、pDC、CD4~+CD25+Treg细胞的百分比以及CD4~+/CD8~+比值相关，有待于进一步研究。更多还原

【Abstract】 OBJECTIVE:①To investigate the relationship between TLR3 in the PBMC of the unbilicalvein blood of babies whose mothers are HBsAg posotive and HBV intrauterineinfection.②To explore the relationship between immunocytes concerning PBMC and theHBV intrauterine infection.③To explore the mechanism of TLR3 in HBV intrauterine infection .METHODS:①The epidemiological base line data and umbilical vein blood of 52HBsAg-positive pregnant women were collected from Taiyuan infectious hospitalfrom July 2011 to January 2012.②Experiment was performed in vitro: The PBMC were separated from umbilicalvein blood and then were cultured. The PBMC of HBV intrauterine infection wereintervented by HBV+PolyI:C and HBV.The PBMC with negative HBV intrauterineinfection were intervented only by HBV.③The protein expession of TLR3 and the percentages of B-cells,T-cells,DC andTreg were detected by FCM.RESULTS:①The expression level of TLR3 protein was higher in PBMC with negativeHBV intrauterine infection than that in PBMC with positive HBV intrauterineinfection(32.18±11.36,30.29±11.04),but no significant difference was observed（t=0.410，P=0.684）.After the intervention of PolyI:C, the expression level of TLR3increased,but still no significant difference was observed (32.18±11.36, 32.16±9.40;t=-1.824,P=0.078).②Proportion of B-cells with negative HBV intrauterine uninfection was higherthan the ones with positive HBV intrauterine infection(7.38±6.40;6.76±4.01),but nosignificant difference was observed (t=0.210,P=0.834).After the intervention ofPolyI:C, percentage of B-cells with positive HBV intrauterine uninfection decreased,but still no significant difference was observed(7.38±6.40,5.14±2.64;t=0.768,P=0.447). ③Percentage of CD4+T-cells with negative HBV intrauterine infection washigher than the ones with positive HBV intrauterine infection,but no significantdifference was observed(41.14±20.08,33.82±2.60;t=0.618,P=0.543),while percentageof CD8+T-cells with negative HBV intrauterine infection was higher than the oneswith positive HBV intrauterine infection(11.30±5.87,11.14±9.64) with no significantdifference(11.30±5.87,11.14±9.64;t=0.041,P=0.968).The difference of ratio ofCD4+T-cells to CD8+T-cells between negative and positive HBV intrauterine infectiongroups was not significant(4.30±2.28,4.75±3.23;t=-0.30,P=0.767).④Percentage of mDC with negaitive HBV intrauterine infection was higher thanthe one with positive HBV intrauterine infection(0.84±0.58,0.67±0.34) but with nosignificant difference (t=-0.485,P=0.633).While percentage of pDC with negaitiveHBV intrauterine infection was higher than the one with positive HBV intrauterineinfection,but with no significant difference there was(0.92±0.91,0.91±0.46;t=0.020,P=0.984).⑤Percentage of CD4+CD25+Treg with negative HBV intrauterine infection washigher than the one with positive HBV intrauterine infection(4.69±3.41,3.00),but withno significant difference between them(t=0.474,P=0.646).CONCLUSIONS:①The fact that expression level of TLR3 protein with negative HBVintrauterine infection was higher than the one with HBV intrauterine infection andthat intervention of PolyI:C caused the increase of the expression level of TLR3protein indicates that HBV intrauterine infection may be linked to TLR3 proteinexpresson.②The fact that the percentages of B-cells,CD4+T-cells and mDC with negativeHBV intrauterine infection were higher than the ones with positive intrauterineinfection indicates that the B-cells,CD4+T-cells,mDC may be a protective factoragainst placenta HBV infection.③The fact that association between percentage of CD8+T-cells,pDC as well asCD4+CD25+Treg and the CD4+/CD8+T-cells was not discovered indicates that furtherstudy is required.更多还原