摘要
Objective To investigate the preventive therapeutic effect and possible mechanism of single chain variable fragments chimeric protein (SD) of ovalbumin epitopes internalizing receptor DEC-205 antibody on food allergy in mice. Methods Mice were randomly divided to five groups (control, PBS, scFv DEC 100 μg, SD 50 μg, SD 100 μg) and treated for 24 hours before OVA administration. After challenge, the serum level of OVA-specific IgE, IgG1, IgG2a and IL-4 were detected by ELISA. Infiltration of eosinophils and mast cells in the jejunum was observed by HE staining and toluidine blue staining respectively. The bone marrow of tibia and femur was isolated and cultured to obtain immature dendritic cells(BMDCs), which were further treated with LPS (10 ng/mL), TSLP (50 ng/mL), scFv DEC protein (1000 ng/mL) and SD protein (10,100,1000)ng/mL for 24 hours, and the IL-10 level of supernatant was assayed by ELISA. Results Compared with PBS group, the number of SD-treated mice with diarrhea was markedly reduced. The difference in rectal temperature and the levels of serum OVA-specific IgE, IgG1, IgG2a and IL-4 decreased significantly after prophylactic administration of SD; The number of eosinophils and mast cells in jejunum also decreased significantly while the IL-10 level in the supernatant of BMDCs increased significantly after SD intervention. Conclusion SD mitigates experimental FA response by fosters the immune tolerance property of dendritic cells.
Subject(s)
Mice , Animals , Ovalbumin , Interleukin-10 , Single-Chain Antibodies/genetics , Immunoglobulin E , Epitopes/therapeutic use , Interleukin-4 , Food Hypersensitivity/prevention & control , Immunoglobulin G , Recombinant Fusion Proteins/genetics , Mice, Inbred BALB C , Disease Models, Animal摘要
Objective To study the effects of B cells from lupus prone Triple congenic (TC) mouse model on the differentiation and development of T cell subsets. Methods The spleen size and B cell numbers were measured, and surface CD40, CD86 and Ⅰ-Ab molecules on B cells as well as CD4+T cell subsets were detected using flow cytometry when the spontaneous systemic lupus erythematosus (SLE) model TC mice and control B6 mice were 6 months old. In addition, the chimera of TC B cells and B6 CD4+T cells or chimera of B6 B cells and B6 CD4+ T cells were transferred into B6.Rag-/- mice via intravenous injection. Then, T cell subsets in the spleen of recipient B6.Rag-/-mice were observed 7 days after cell transplantation. Results TC mice had significantly bigger spleen [(5337±934) mg] and more CD19+B cell number [(276.0±48.7)×107] than control B6 mice [spleen weight: (91±4) mg; B cell number: (6.4±0.3)×107](P0.05). The recipient B6.Rag-/-mice transplanted with TC B cells had significantly more Th1 subset [(54.1±2.8)%] and IL-21+CD4+T cell population [(14.3±1.0)%], but less Th17 subset [(2.05±0.09)%] in spleen than the recipient B6.Rag-/-mice administered by B6 B cells [Th1 subset: (39.5±1.1)%; IL-21+CD4+T cell population:(7.5±1.2)%;Th17 subset:(6.45±1.10)%](P<0.01). Conclusion The B cells of lupus-prone TC mice exhibit a markedly hyper-activation in spleen, and promote CD4+T cells differentiation preferentially into Th1 subset and IL-21+CD4+T cell population, which may further contribute to SLE pathogenesis.
摘要
Aim To investigate the effect of PLK1 on epithelial-mesenchymal transition (EMT)of human e-sophageal squamous cell carcinoma (ESCC)cells TE-1 5 and its relevant molecular mechanisms.Methods PLK1 overexpressed ESCC cells and control vector were used as the experimental cells.The expression of EMT-related protein markers E-cadherin and vimentin were measured by Western blot.vimentin mRNA was measured by Real-time PCR.Total cellular protein and nuclear protein were respectively extracted,and then they were used to detect the expression of β-catenin by Western blot.β-catenin siRNA and non-specific siR-NA were transiently transfected into the cell clones overexpressed PLK1 ,and then vimentin was detected by Western blot.β-catenin protein degradation com-plex was detected by immunoprecipitation and Western blot.Results The mesenchymal marker vimentin was distinctively upregulated and the epithelial marker E-cadherin was distinctively downregulated in the cell clones overexpressed PLK1 ,compared with those in the vector clones.This indicated that EMT occurred in ESCC cells.vimentin mRNA was also markedly in-creased.In the cell clones overexpressed PLK1 ,β-catenin were both elevated from the total cells and the nucleus.The expression of vimentin was reduced whenβ-catenin was knocked down.APC and GSK-3βwere both reduced from Axin immunoprecipitate in the cell clones overexpressed PLK1 .Conclusion PLK1 up-regulates vimentin and promotes EMT in ESCC cells probably by inhibiting the formation of protein degrada-tion complex and stabilizing β-catenin.