As a T cell-mediated autoimmune disease,type 1 diabetes mellitus(T1DM)is marked by insulin defect resulting from the destruction of pancreaticβ-cells.The understanding of various aspects of T1DM,such as its epidemiol...As a T cell-mediated autoimmune disease,type 1 diabetes mellitus(T1DM)is marked by insulin defect resulting from the destruction of pancreaticβ-cells.The understanding of various aspects of T1DM,such as its epidemiology,pathobiology,pathogenesis,clinical manifestations,and complications,has been greatly promoted by valuable research performed during the past decades.However,these findings have not been translated into an effective treatment.The ideal treatment should safely repair the destroyed immune balance in a longlasting manner,preventing or stopping the destruction ofβ-cells.As a type of immune hypo-responsiveness to the orally administrated antigen,oral tolerance may be induced by enhancement of regulatory T cells(Tregs)or by anergy/deletion of T cells,depending on the dosage of orally administrated antigen.Acting as an antigen-specific immunotherapy,oral tolerance therapy for T1DM has been mainly performed using animal models and some clinical trials have been completed or are still ongoing.Based on the review of the proposed mechanism of the development of T1DM and oral tolerance,we give a current overview of oral tolerance therapy for T1DM conducted in both animal models and clinical trials.展开更多
Analysis of the frequency of antigen-specific cytotoxic T lymphocytes (CTLs) ex vivo is largely dependent on the use of MHC/peptide tetramers. However, the latter reagents have not been widely available, most likely b...Analysis of the frequency of antigen-specific cytotoxic T lymphocytes (CTLs) ex vivo is largely dependent on the use of MHC/peptide tetramers. However, the latter reagents have not been widely available, most likely because of their costly and time-consuming production. In this report we utilized an economic strategy to construct HLA/peptide tetramers with recombinant peptide-linked β2 microglobulin (β2m). The HLA-A2-restricted, melanoma antigen MARTI-derived pep- tide MART127-35( AAGIGILTV) was fused to the N terminus of human β2m through a 15-amino acid (aa)-long linker before being refolded with the recombinant biotinylated HLA-A2 heavy chain ectodomain. The resulted 2-component (2C) monomer was then tetramerized with phycoerythin-labeled streptavidin. The experimental result showed that the 2C HLA-A2/ MART127-35 monomer was shown to bind to the HLA class complex-specific monoclonal antibody W6/32 and the HLA-A2/ MART127-35 complex-specific single chain antibody fragment (scFv) 8.3, suggesting the correctness of its specificity. Fur- thermore, the 2C HLA-A2/MART127-35 tetramer detected a specific CD8+ T cell population in HLA-A2-restricted melanoma infiltrating lymphocytes as the conventional 3C HLA-A2/MART127-35 tetramer. The yield of 2C HLA-A2/MART127-35 monomer was 2. 5 times more than that of the conventional 3C monomer. Taken together, these data indicate that the HLA-A2/ MART127-35 tetramer can be generated conveniently through the use of MART127-35 peptide-β2 m fusion proteins, which can fa- cilitate the monitoring of HLA-A2-restricted, MART1-specific CTL responses in patients with melanoma.展开更多
基金Supported by National Natural Science Foundation of China,No.81803418Natural Science Foundation of the Jiangsu Higher Education Institutions,No.18KJD350001and Project for Youth Scholar of Jiangsu Collaborative Innovation Center of Regional Modern Agriculture and Environmental Projection,No.HSXT2-314.
文摘As a T cell-mediated autoimmune disease,type 1 diabetes mellitus(T1DM)is marked by insulin defect resulting from the destruction of pancreaticβ-cells.The understanding of various aspects of T1DM,such as its epidemiology,pathobiology,pathogenesis,clinical manifestations,and complications,has been greatly promoted by valuable research performed during the past decades.However,these findings have not been translated into an effective treatment.The ideal treatment should safely repair the destroyed immune balance in a longlasting manner,preventing or stopping the destruction ofβ-cells.As a type of immune hypo-responsiveness to the orally administrated antigen,oral tolerance may be induced by enhancement of regulatory T cells(Tregs)or by anergy/deletion of T cells,depending on the dosage of orally administrated antigen.Acting as an antigen-specific immunotherapy,oral tolerance therapy for T1DM has been mainly performed using animal models and some clinical trials have been completed or are still ongoing.Based on the review of the proposed mechanism of the development of T1DM and oral tolerance,we give a current overview of oral tolerance therapy for T1DM conducted in both animal models and clinical trials.
文摘Analysis of the frequency of antigen-specific cytotoxic T lymphocytes (CTLs) ex vivo is largely dependent on the use of MHC/peptide tetramers. However, the latter reagents have not been widely available, most likely because of their costly and time-consuming production. In this report we utilized an economic strategy to construct HLA/peptide tetramers with recombinant peptide-linked β2 microglobulin (β2m). The HLA-A2-restricted, melanoma antigen MARTI-derived pep- tide MART127-35( AAGIGILTV) was fused to the N terminus of human β2m through a 15-amino acid (aa)-long linker before being refolded with the recombinant biotinylated HLA-A2 heavy chain ectodomain. The resulted 2-component (2C) monomer was then tetramerized with phycoerythin-labeled streptavidin. The experimental result showed that the 2C HLA-A2/ MART127-35 monomer was shown to bind to the HLA class complex-specific monoclonal antibody W6/32 and the HLA-A2/ MART127-35 complex-specific single chain antibody fragment (scFv) 8.3, suggesting the correctness of its specificity. Fur- thermore, the 2C HLA-A2/MART127-35 tetramer detected a specific CD8+ T cell population in HLA-A2-restricted melanoma infiltrating lymphocytes as the conventional 3C HLA-A2/MART127-35 tetramer. The yield of 2C HLA-A2/MART127-35 monomer was 2. 5 times more than that of the conventional 3C monomer. Taken together, these data indicate that the HLA-A2/ MART127-35 tetramer can be generated conveniently through the use of MART127-35 peptide-β2 m fusion proteins, which can fa- cilitate the monitoring of HLA-A2-restricted, MART1-specific CTL responses in patients with melanoma.
