| OBJECTIVE:①To explore the role of TLR3in HBV intrauterine transmission of newborns who had HBsAg positive mothers.②To explore the relationship between TLR3associated immune cells and HBV intrauterine transmission.③To understand the immune status of HBV intrauterine transmission of newborn whose mothers were HBsAg positive.METHODS:①320HBsAg-positive pregnant women and their newborns were collected from department of gynecology and obstetrics in Third People’s Hospital of Taiyuan from July2011to January2012. Information were obtained by face to face questionnaire survey.②3ml femoral venous blood of newborns with anticoagulant and without anti-coagulant were collected24h after birth. HBV markers and HBV DNA in peripheral blood of HBsAg-positive pregnant women and newborns were detected by ELISA and FQ-PCR respectively.③PBMCs in a part of femoral venous blood of newborns with anticoagulant were separated by density gradient centrifugation. The expression level of TLR3on PBMCs was assayed by multicolor flow cytometry (FCM) Count and percentage of dendritic cells, B cells, CD4+CD25+Treg and CD4+CD8+T lymphocyte of the rest were detected by fluorescence-activated cell sorting (FACS) Then the relationship between TLR3, related immune cells and HBV intrauterine transmission was analysised.RESULTS:①Among320newborns whose mothers were HBsAg-positive,45cases happened HBV intrauterine transmission,39newborns were only HBsAg-positive,6newborns were HBsAg-positive and HBV DNA content≥103copies/mL, HBV intrauterine transmission rate was14.06%(45/320).②In HBV intrauterine transmission group, the average age of HBsAg-positive mothers was25.09±3.44years old, the average gestational age was39.37±1.16weeks, newborns birth weight was3307.11±410.13g, the birth of average height was50.19±1.81cm, the ratio of male and female newborn was1:1.14(21:24), the ratio of vaginal delivery and caesarean was1:4.63(8:37),11cases were HBV DNA content <500copies/ml;25cases were HBV DNA content>107copies/ml,9cases were of HBV DNA load content≤107copies/ml. In HBV no intrauterine transmission group, the average age of HBsAg-positive mothers was27.27±3.60years old, the average gestational age was39.31±1.12weeks, newborns birth weight was3395.18±423.50g, the birth of average height was50.07±1.39cm, the ratio of male and female newborn was1:1.17(127:149), the ratio of vaginal delivery and caesarean was1:1.12(130:146),165cases were HBV DNA content<500copies/ml;72cases were HBV DNA content>107copies/ml,9cases were of HBV DNA load content≤107copies/ml. Compared with intrauterine infection group and non intrauterine infection group, analysis by t test, there is significant difference in age of HBV intrauterine infection group and non pregnant women with intrauterine transmission between groups (t=-3.895, P<0.001), analysis by χ2test, the difference was statistically significant neonatal intrauterine transmission of HBV and HBsAg positive way of childbirth (χ2=24.439, P<0.001); and between the two groups in HBsAg positive pregnant women and newborn sex difference was not statistically significant (P>0.05). ③To analysis of the relationship between HBV inflectional status of HBsAg positive mothers and HBV intrauterine transmission, the results showed HBsAg positive pregnant women’HBsAg titer was statistically relative to HBV intrauterine transmission rate (P<0.001), the HBV intrauterine transmission incidence of mothers be HBsAg<3000ng/mL was22%(37/168), intrauterine transmission rate of mothers be HBsAg>3000ng/mL was5.3%(8/152); intrauterine transmission rate of mothers be HBsAg (+) HBeAg (-) was7%, intrauterine transmission rate of mothers be HBsAg (+) HBeAg (+) was increased to22.3%as HBeAg titers increasing (P<0.001); HBV intrauterine transmission rate(24%) of HBsAg (+) anti-HBe (+)mothers was higher than that of mothers be HBsAg (+) anti-HBe (-)(P<0.001); intrauterine transmission rate(26.3%) of mothers be HBV DNA>105copies/ml was higher than that of mothers be103-105HBV DNA copies/ml and HBV DNA (-)(P<0.001); anti-HBs and anti-HBe titer values of mothers were not statistically relative to HBV intrauterine transmission (P>0.05).④The results of multiple-factor analysis showed that maternal HBeAg level (OR=1.001,95%CI:1.001～1.002) and maternal age (OR=0.840,95%CI:0.757～0.934) were associated with HBV intrauterine transmission.⑤The mean fluorescence intensity of TLR3(27.66±11.89)%in intrauterine transmission group was higher than that of non-intrauterine transmission group (20.06±8.59)%, but there were no significant difference between two groups (t=0.155,P=0.877).⑥Both mDC cells ratio(1.09±1.16)%and pDC cells ratio(0.95±0.65)%of femoral venous blood in non intrauterine transmission group was higher than that of (0.95±0.65)%and (1.09±1.16)%in intrauterine transmission group, but there were no significant difference between two groups (t=-0.544,P=0.587; t=-0.544, P=0.587).⑦Bcells ratio(6.91±4.49)%of newborns femoral venous blood in non intrauterine transmission was higher than that of (6.86±4.30)%in intrauterine transmission group, but there were no significant difference between two groups (t=-0.057, P=0.954).⑧CD3+T cell ratio (60.28±14.50)%, CD4+Tcell ratio (42.26±11.02)%, CD8+T cell ratio (15.27±5.93)%of femoral venous blood in intrauterine transmission group were higher than CD3+T cell (58.52±16.23)%, CD4+T cell (39.92±14.52)%, CD8+T cell (15.27±5.93)%of non intrauterine transmission group, but there were not statistical difference between two groups (t=0.478, P=0.662; t=0.804, P=0.442; t=0.350, P=0.727). CD4+/CD8+ratio (3.44±1.68) in intrauterine transmission group was lower than that of (3.34±1.92) non intrauterine transmission group, but there was no significant difference between two groups(t=0.203, P=0.839).⑨CD4+CD25+FOXP3Treg ratio (4.62±4.19)%of newborns femoral venous blood in HBV intrauterine transmission group was lower than that (11.79±14.25)%of non intrauterine transmission group(t=-0.638, P=0.525). CD4+FOXP3+Treg ratio(4.22±2.28)%and CD4+CD25+Treg ratio(10.01±5.05)%of newborns femoral venous in intrauterine transmission group were lower than CD4+FOXP3+Treg (4.64±5.59)%and CD4+CD25+Treg ratio(11.79±14.25)%of non intrauterine transmission group, but there were no significant difference between two groups (t=-0.339, P=0.735; t=-0.568, P=0.570).CONCLUSIONS:①The study found that age and HBeAg titer of HBsAg positive mothers had influence on newborn HBV intrauterine transmission.②TLR3protein content of intrauterine transmission group was higher than that of non intrauterine transmission group. This suggested that TLR3may play an important role in HBV intrauterine transmission. ③B cells ratio of newborn femoral venous blood in non intrauterine transmission is higher than that of intrauterine transmission group. This suggested that hum oral immune response of newborn may be related to HBV intrauterine transmission.④mDC cells and pDC ratio cells of non intrauterine transmission group ratio were higher than that of intrauterine transmission group. This suggested that DC ratio of newborns of femoral venous blood may be related to HBV intrauterine transmission.⑤Intrauterine transmission group of newborn CD3+, CD4+, CD8+T lymphocyte ratio and the ratio of CD4+/CD8+T cell subsets in non intrauterine transmission group was higher than that of the newborn. Intrauterine transmission of may in HBV infection, no effective immune response, resulting in intrauterine transmission.⑥Non HBV intrauterine transmission of CD4+CD25+FOXP3+Treg is relatively higher proportion of intrauterine transmission group, probably due to HBsAg positive mother newborn immune cells play a negative regulatory role, HBV intrauterine transmission might be reduced the relative proportion of cells... |
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