Relationship Between Myeloid-derived Suppressor Cells And Biliary Atresia

Objectives1. To detect the expression of myeloid derived suppressor cells in the peripheralblood of biliary atresia patients and investigate the relationship between myeloidderived suppressor cells and pathology classification.2. To explore the correlation between MDSCs and11kinds of cytokines in biliaryatresia，to explore the functions of MDSCs in biliary atresia.MethodsPeripheral blood samples were collected from BA patients (n=27), nonhepaticcongenital anomalies patients (n=7), CBD patients (n=27), NH patients (n=13), andthe expression of MDSCs (CD33+CD11b+HLA-DR-) from peripheral blood sampleswere detected by flow cytometry. At the same time, the serum blood samples fromBA (n=18), CBD (n=13), NH (n=8) patients were collected, the expression of11kinds of cytokines were tested by flowing microsphere technology. Meanwhile,Samples of liver tissues were ground and then supernatants were harvested,11kindsof cytokines were tested. The formalin-fixed paraffin liver specimens were cut intopieces and subsequently performed with pathological and immunohistochemicalstaining. Immunohistochemical staining were carried out by using double stainingmethod to find MDSCs (CD33+HLA-DR-). Results1. The expression of MDSCs in whole bloodThe frequency of MDSCs (CD33+CD11b+HLA-DR-) in whole blood of BApatients was significantly increased in comparison with controls and CBD patients,although there is no difference between BA and NH patients. Moreover, MDSCslevels correlate with pathological classification of BA, the late stage was significantlyincreased when compared with the early stage.2. The pathological changes of biliary atresia and immunohistochemicalstaining of MDSCsBile duct hyperplasia, degeneration, varying degrees of fibrosis could be found inthe periportal area, meanwhile, multinucleated giant cells and pseudolobule could befound in the liver tissue of the patients. Lymphocytes and granulocyte were infiltratedaround the bile canalicus, with various degrees of fiber bundle package timid tube.MDSCs were found around bile canalicus in liver tissue samples of BA.3. The expression of11kinds of cytokines in serum and liver tissueAmong the11kinds of cytokines,the expression of IFN-γ、IL-2、IL-10、IL-8、IL-6、IL-1β and TNF-β were significantly increased in serum blood of BA. The levelof IFN-γ was lower in liver tissues of BA than in the control groups, however, thelevel of IL-4was higher in liver tissues of BA than in the control groups.4. The correlation between MDSCs and cytokinesThe expression of MDSCs show strong positive correlation with some kinds ofcytokines（IL-1β, TNF-β）.Conclusions1. MDSCs has a close relationship with biliary atresia, MDSCs levels correlate withpathological classification of BA 2. The increase of MDSCs may be correlated with T cell cytokines.