ABSTRACT
Objective To investigate the clinical features and diagnosis and treatment of acute fatty liver of pregnancy(AFLP). Methods A retrospective analysis was performed for the clinical data of 12 patients with AFLP who were diagnosed and treated in Department of Infec-tious Diseases,Tongji Hospital,Tongji Medical College of Huazhong University of Science and Technology,from April 2012 to March 2017, including general data,clinical manifestations,laboratory markers,imaging examinations,treatment,and prognosis. Results All 12 pa-tients developed AFLP in late pregnancy,and major clinical manifestations included gastrointestinal symptoms,liver failure,jaundice,and coagulation disorder. All patients were given multimodality therapy to protect the liver,improve coagulation,and reduce infection;11 pa-tients underwent cesarean section;6 underwent blood filtration;5 underwent plasma exchange. One patient died,resulting in a mortality rate of 8.3%;5 perinatal infants died,resulting in a mortality rate of 35.7%. Conclusion In patients with AFLP,early diagnosis,timely ter-mination of pregnancy,maximum symptomatic/supportive treatment,and control of infection,as well as the artificial liver support system,is the key to improving the prognosis of mothers and infants.
ABSTRACT
CD4(+)CD25(+)CD127(dim/-) regulatory T cells (Tregs) have been implicated in suppressing T cell immune responses to hepatitis B virus (HBV), but the inhibition mechanism has not being clear yet. This study investigated the effects of soluble FGL2 (sFGL2) secreted by Tregs on immune suppression in chronic HBV-infected patients. We verified that sFGL2 protein and mRNA were highly expressed in Tregs. The separated Tregs by using magnetic beads from peripheral blood mononuclear cells (PBMCs) in 20 patients with chronic hepatitis B were co-cultured with PBMCs at a ratio of 1:3 with anti-CD3 stimulating antibody or FGL2 blocking antibody. The proliferation index of CD8(+)T cells after blocking FGL2 was higher than that in blank group (3.58±0.18 vs. 3.28±0.17, P=0.034) in 18 of 20 samples, and lower than that in CD3 stimulation group (3.82±0.19, P=0.026) in 16 of 20 samples. The IFN-γ secreted in the mixed culture in the absence of Tregs was higher than that in the culture in the presence of Tregs, but it could be abolished by FGL2 blocking antibody. These results suggest that sFGL2 protein secreted by Tregs suppresses the proliferation and function of CD8(+) T cells in chronic hepatitis B.
ABSTRACT
CD4(+)CD25(+)CD127(dim/-) regulatory T cells (Tregs) have been implicated in suppressing T cell immune responses to hepatitis B virus (HBV), but the inhibition mechanism has not being clear yet. This study investigated the effects of soluble FGL2 (sFGL2) secreted by Tregs on immune suppression in chronic HBV-infected patients. We verified that sFGL2 protein and mRNA were highly expressed in Tregs. The separated Tregs by using magnetic beads from peripheral blood mononuclear cells (PBMCs) in 20 patients with chronic hepatitis B were co-cultured with PBMCs at a ratio of 1:3 with anti-CD3 stimulating antibody or FGL2 blocking antibody. The proliferation index of CD8(+)T cells after blocking FGL2 was higher than that in blank group (3.58±0.18 vs. 3.28±0.17, P=0.034) in 18 of 20 samples, and lower than that in CD3 stimulation group (3.82±0.19, P=0.026) in 16 of 20 samples. The IFN-γ secreted in the mixed culture in the absence of Tregs was higher than that in the culture in the presence of Tregs, but it could be abolished by FGL2 blocking antibody. These results suggest that sFGL2 protein secreted by Tregs suppresses the proliferation and function of CD8(+) T cells in chronic hepatitis B.
Subject(s)
Humans , CD8-Positive T-Lymphocytes , Allergy and Immunology , Metabolism , Cells, Cultured , Fibrinogen , Allergy and Immunology , Metabolism , Hepatitis B, Chronic , Allergy and Immunology , Metabolism , T-Lymphocytes, Regulatory , Allergy and Immunology , MetabolismABSTRACT
To obtain single chain variable fragment (scFv) and bivalent single chain variable fragment (bsFv) against transferrin receptor, up-stream and down-stream primers were designed according to the complementary sequences of FR1 region of variable heavy (VH) and FR4 of variable light (VL), respectively, which contained inter-linker G4S and the restriction endonuclease SfiI, AscI and NotI. Two pieces of scFv fragments were first amplified through PCR and then inserted into plasmid pAB1, which could express scFv protein once induced by IPTG in the host bacteria. To express scFv and bsFv, E. coli TG1 was cultured in LB broth and was induced by IPTG. The restriction enzyme digestion map and DNA sequencing demonstrated that scFv and bsFv genes were successfully inserted into the expression plasmid. SDS-PAGE and Western blotting revealed the protein band at 35kD and 60kD, which were consistent with the molecular weight of scFv and bsFv respectively. Flow cytometry showed that scFv and bsFv harbored the specific binding activity with TfR expressed in various tumor cells, and the avidity of bsFv was higher than that of the parent scFv.