Investigation Of Maternal Microchimerism Impact Correlated In The Pathogenesis Of Biliary Atresia

ObjectivesDuring the period of pregnancy,the immunological competence was in suppression to prevent the rejection of fetus,which made it possible that the bilateral exchange of cells between mothers and fetus becomed the major mechanism of bringing about microchimerism. Microchimeric cells maybe the cause of some autoimmune disease, and it was reported that immuno-competent maternal cells could cause GVHD-like symptoms in patients with severe combined immuno deficiency,whose manifestations were similar with biliary atresia. In our research,the correlation between maternal microchimerism and biliary atresia was investigated.Material and MethodsTwenty cases of paraffin-embedded liver tissues were involved, each tissue was cut consecutively into 6 sections, single 4μm thick. Exam the tissue morphology under light microscope for target site. At 0.5-minute intervals, enzymatic digestion was carried out in 5 sections for 2-4 minutes. After DAPI dying sufficiently, evaluate target integrity digestion of every tissue section under fluorescence microscope to determine an optimal time, following which to digest the sixth section, and then hybridizate with FISH probes. Analyse the signals and count after posthybridization washes. By which mentioned above , the optimization of digestion pretreatment and signals interpretation in fluorescence in situ hybridization technique on paraffin-embedded tissues was finished. Using the chromosome enumeration probes of fluorescent in situ hybridization(FISH), the analysis of X and Y chromosomes were performed on liver paraffin sections of 12 male BA cases and 10 male controls, which involved 8 neonatal hepatitis and 2 choledochus cyst cases. Counted the maternal cells with XX chromosomes in each liver section.ResultsUnder fluorescence microscope, the tissue morphology for an optimal digestion time is effective DAPI blue uptake and well-defined, intact cell peripheries (as viewed using a DAPI filter), as well as consecutive borders green and clear outlines (as viewed using a FITC filter). Digestion time evaluation of heterogeneous tissue section was based on target site of the whole section. Independent cells adapt for analysis should be well-defined, DAPI staining uniform and signals distinct. Maternalcells with XX chromosomes were found in liver sections of all male BA cases, while occasionally in male control cases. The maternal cells in per 30 views appropriate of BA group and control group were 11±2.59 and 1.8±1.69, respectively(P<0.01), demonstrating a significant statistical difference.Conclusions1. Maternal microchimerism could be found in the liver tissues of both BA and other infant hepatopathy cases;2.The quantity of maternal cells in the male BA livers was significantly higher than that in other neonatal liverdiseases;3.Consequently, it is possible that maternal microchimerism contributes to the pathogenesis of BA.