【作者】 马丽娟；

【导师】 马玉燕；

【作者基本信息】 山东大学 ， 妇产科学， 2008， 硕士

【摘要】 第一部分:胎儿HBeAg与乙型肝炎病毒宫内感染关系的研究研究背景及目的孕妇及新生儿乙肝病毒e抗原(HBeAg)阳性被认为是发生HBV宫内感染的高危因素,但HBeAg是否直接参与HBV宫内感染的发生尚无定论。本研究旨在通过检测HBeAg对胎儿免疫机制的影响来探讨探讨HBeAg在胎儿HBV宫内感染中所起作用。方法将HBsAg阳性孕妇分娩的新生儿50例分为两组:HBeAg阳性组(n=23)和HBeAg阴性组(n=27),分别采用双抗夹心酶联免疫免疫吸附法(DAS-ELISA)和病毒核酸扩增法(实时免疫荧光定量PCR方法)对孕妇静脉血和新生儿脐带血进行HBV血清学标志物和病毒含量的检测,采用DAS-ELISA法检测新生儿脐血中细胞因子白细胞介素—2(IL-2)、干扰素—γ(IFN-γ)、白细胞介素—4(IL-4)和白细胞介素—6(IL-6)水平,比较两组之间细胞因子水平的差异,同时将HBsAg阴性孕妇分娩的新生儿作为正常对照组。结果孕妇HBeAg阳性和阴性者分别为27例和23例,其新生儿脐血HBeAg阳性者分别为22例(81.48%)和1例(4.35%),HBeAg阳性孕妇的新生儿HBeAg阳性率显著高于HBeAg阴性孕妇的新生儿,差异具有显著性(P＜0.05);HBeAg阳性的新生儿(n=23)脐血IL-2、IFN-γ含量显著低于HBeAg阴性组(n=27)(26.08±5.12pg/ml vs 32.16±6.26pg/ml、12.14±1.89pg/ml vs17.20±5.39pg/ml,P＜0.05),而IL-4、IL-6水平则显著高于HBeAg阴性组(17.35±3.18pg/ml vs 11.03±2.64pg/ml、24.01±4.12pg/m vs 17.91±2.81pg/ml);正常对照组脐血IL-2、IFN-γ、IL-4、IL-6水平分别为(33.39±9.69pg/ml、19.27±3.80pg/ml、12.77±4.29pg/ml、18.35±3.24pg/ml),与HBeAg阴性组之间比较无统计学差异(P＞0.05)。结论胎儿体内的HBeAg可由母体传播而来并引起胎儿机体免疫失衡,致使机体不能及时有效地清除病毒,并且可导致胎儿免疫耐受,与HBV宫内感染的发生有密切关系。第二部分:CD35在胎盘细胞表达的研究研究背景及目的胎盘感染是引起母婴宫内垂直传播的高危因素,胎盘上的乙型肝炎病毒(HBV)是以HBsAg-抗HBs-补体C3复合物的形式存在的,这种复合物可沉积在滋养层细胞表面与Ig-G受体或者补体C3的受体结合而导致病毒在胎盘上的逐层传递,这对新生儿HBV宫内感染有重要意义;现有研究表明,胎盘组织上固有表达FcγRⅢ受体,可能对HBV宫内传播起到重要作用,但未有研究证实胎盘组织表达补体C3的受体的情况。补体C3的受体为补体受体1型CR1,即CD35,本试验通过检测在胎盘上CD35的表达情况,进一步明确HBV经胎盘途径传播的机制。方法选择HBsAg阳性孕妇6例,HBsAg阴性孕妇3例,剖宫产中取胎盘组织进行石蜡切片,采用用免疫组织化学SABC方法检测胎盘组织中CD35的表达。结果切片免疫组化染色显示CD35在胎盘组织血管中的红细胞及白细胞上表达呈阳性,而在胎盘组织滋养层和间质细胞中均未发现CD35的表达。结论胎盘组织不存在补体C3的受体,因此HBV感染胎盘时,抗原抗体补体复合物不是通过与补体C3的受体的结合而介导胎儿宫内传播的。更多还原

【Abstract】 Part One:Study on the relationship between HBeAg of fetus and Intrauterine vertical transmission of hepatitis B virusAbstract:Background and Objective HBeAg of the pregnant women and neonates is a high risk factor of Intrauterine vertical transmission of HBV,but it is unknown that if HBeAg is the direct reason of intrauterine infection of HBV.Our research is to study the mechanism of maternal-infant vertical transmission of hepatitis B virus by testing the influence of the HBeAg on the immunologic state of neonates.Methods 50 neonates born from positive HBsAg mother were divided into positive HBeAg group(n=23)and negative HBeAg group(n=27).By using diantibody sandwich enzyme linked immunosorbant assay(DAS-ELISA)and polymerase chain reaction(PCR),the serum HBV M and HBV DNA were detected in these pregnant women and neonates,IFN-γ,IL-2,IL-4 and IL-6 in these neonates were also detected by using DAS-ELISA,and theses cytokine levels were compared between the two groups,the control group(20 neonates born from negative HBsAg mothers)was set meanwhile.Results Pregnant women were divided into HBeAg group(27 cases)and negative HBeAg group(23 cases),the cases of positive HBeAg neonates of each group were 22(81.48%)and 1(4.35%),The number of positive HBeAg neonates born from positive HBeAg mothers was much more than it is of the positive HBeAg neonates born from negative HBeAg mothers,which had significant difference(P＜0.05);The IL-2,IFN-γlevels of the positive HBeAg group neonates were significant lower than those of the negitive HBeAg group(26.08±5.12pg/ml vs 32.16±6.26pg/ml,12.14±1.89pg/ml vs 17.20±5.39pg/ml),the IL-4,IL-6 levels were signifinant higher 17.35±3.18pg/ml vs 11.03±2.64pg/ml,24.01±4.12pg/ml vs 17.91±2.81pg/ml),and those of the control group were(33.39±9.69pg/ml,19.27±3.80pg/ml, 12.77±4.29pg/ml,18.35±3.24pg/ml),which had no significant difference with the nagative HBeAg group(P＞0.05).Conclusion The HBeAg of positive HBsAg pregnant woman can be transmitted into her neonates and causes immunity unbalance,which makes the organism can not clear the HBV in time and efficiently and brings on the immunotolerance of hepatitis B virus meanwhile,and these mechanisums above-mentioned have the close relationship with HBV maternal-infant vertical transmission. Part two:study on the distribution in human placentaAbstract:Background and Objective The placental tissue infection is a high risk factor of the occurrence of HBV intrauterine vertical transmission,the former researches show that the HBV in the placenta exist by means of HBsAg-anti-HBs-C3c complex,the later can attached to the cytoplasm of trophobasts and combined with the related acceptor of Ig-G or C3,which has the important significance of neonates’ intrauterine infection;some studies discovered that FcγRⅢexisted in the placenta,which might be very important to the HBV intrauterine transmission,but there is no research to show that if there is acceptor of C3c in the placenta.The acceptor of C3 is CR1(CD35).Our experiment is to study the research of the HBV placenta infection by testing the expression of CD35 on the placenta.Methods 6 negative pregnant women and 3 normal women were studied in our study,placenta of which were detected by immunohistochemistry SABC to show the situation of CD35expression on placenta.Results the result showed that CD35 expressed on the blood cells of placenta blood vessels,but there was no CD35 discovered in placenta trophoblastic cells and interstitial cells.Conclusion CD35 does not exist in placenta and can not cause the HBV intrauterine infection by combining with the HBsAg-anti-HBsAg-C3c complex.更多还原