AIM： To investigate the mechanism underlying the loss of responsiveness to anti-vascular endothelial growth factor（VEGF） treatment after repeated injections for choroidal neovascularization, VEGF and VEGF receptor...AIM： To investigate the mechanism underlying the loss of responsiveness to anti-vascular endothelial growth factor（VEGF） treatment after repeated injections for choroidal neovascularization, VEGF and VEGF receptor（VEGFR） expressions were evaluated following repeated bevacizumab treatments in hypoxic human umbilical vein endothelial cells（HUVECs） in vitro.METHODS： HUVECs were incubated under hypoxic conditions in two media of different bevacizumab concentrations（1.0 or 2.5 mg/m L） for 17 h, and then in a new medium without bevacizumab for 7h. This procedure was repeated twice more. A culture with an identical volume of excipients served as the control. Cytotoxicity and cell proliferation were assessed using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide and Ki-67 assays, respectively. Levels of VEGF and VEGFR were assessed using enzyme-linked immunosorbent assay and Western blot respectively.RESULTS： Cytotoxic effects were not reported for either bevacizumab concentration. Cell proliferation was not reduced after anti-VEGF treatments. VEGF level after single treatment was significantly higher than that of the control and after repeated treatments. Phosphorylated VEGFR-2 expression increased significantly after singleand repeated bevacizumab treatments compared with the control. The 1.0 mg/m L bevacizumab induced significantly higher expressions of VEGFR-2 than the 2.5 mg/m L in single and repeated treatment groups.CONCLUSION： Bevacizumab treatment of HUVECs elevated VEGFR expression in both single and repeated treatments, indicating a mechanism for the reduced efficacy of anti-VEGF therapy in ocular neovascular disorders.展开更多
The antitumor capabilities of agonistic anti-4-1BB mAbs have made them an attractive target for tumor immunotherapy.However,the adverse side effects associated with agonist antibodies have hindered their clinical deve...The antitumor capabilities of agonistic anti-4-1BB mAbs have made them an attractive target for tumor immunotherapy.However,the adverse side effects associated with agonist antibodies have hindered their clinical development.Here,we aimed to study the immune-related adverse events of repeated doses and long-term use of agonistic anti-4-1BB mAbs.We show that chronic activation of 4-1BB signals induced the accumulation of IFN-γ-producing PD-1^(+)CD8^(+)T cells in the secondary lymphoid organs of tumor-bearing mice by increasing the number of dividing CD8^(+)T cells,which was beneficial for suppressing tumor growth in the early phase of anti-4-1BB induction.However,repeated exposure to anti-4-1BB mAbs led to granuloma development in tumor-draining lymph nodes(TDLNs)of mice due to recruitment and accumulation of macrophages via the CD8^(+)T cell-IFN-γaxis.This was accompanied by excessive lymph node swelling,which impaired the sequential activation of CD8^(+)T cells.Our data provide insights into the immune-related adverse events of long-term agonist 4-1BB antibody dosing,which should be considered during the clinical development of immunomodulating therapy.展开更多
4-1BB is an inducible receptor expressed on activated T cells,while its ligand,4-1BBL,is mainly expressed in antigen-presenting cells and macrophages[1,2].To the best of our knowledge,ligandmediated transactivation of...4-1BB is an inducible receptor expressed on activated T cells,while its ligand,4-1BBL,is mainly expressed in antigen-presenting cells and macrophages[1,2].To the best of our knowledge,ligandmediated transactivation of 4-1BB is responsible for the survival and immune effector functions of T cells.展开更多
Reading biomolecular signatures and understanding their role in health and disease is one of the greatest scientific challenges in modern biology.Decoding this information is not only foundational for biology but also...Reading biomolecular signatures and understanding their role in health and disease is one of the greatest scientific challenges in modern biology.Decoding this information is not only foundational for biology but also a cornerstone for next generation molecular diagnostics.This calls for novel methods that can capture the presence and identity of low-abundance compounds at the individual molecule level.Since its inception in the 1980s,nanopore sequencing has become an essential part of the single-molecule sensing toolkit,proving that long,labelfree reads of DNA can be achieved at low cost and high throughput.1 Despite their huge success in genome sequencing,reading the linear sequence of proteins is a considerably more complex task that requires differentiating 20 different amino acids(as opposed to the 4 DNA bases),as well as their modifications,during real-time translocation.Deconvoluting the time-dependent electrical current traces to determine entire amino acid sequences over long reads is as yet an unaccomplished milestone.展开更多
文摘AIM： To investigate the mechanism underlying the loss of responsiveness to anti-vascular endothelial growth factor（VEGF） treatment after repeated injections for choroidal neovascularization, VEGF and VEGF receptor（VEGFR） expressions were evaluated following repeated bevacizumab treatments in hypoxic human umbilical vein endothelial cells（HUVECs） in vitro.METHODS： HUVECs were incubated under hypoxic conditions in two media of different bevacizumab concentrations（1.0 or 2.5 mg/m L） for 17 h, and then in a new medium without bevacizumab for 7h. This procedure was repeated twice more. A culture with an identical volume of excipients served as the control. Cytotoxicity and cell proliferation were assessed using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide and Ki-67 assays, respectively. Levels of VEGF and VEGFR were assessed using enzyme-linked immunosorbent assay and Western blot respectively.RESULTS： Cytotoxic effects were not reported for either bevacizumab concentration. Cell proliferation was not reduced after anti-VEGF treatments. VEGF level after single treatment was significantly higher than that of the control and after repeated treatments. Phosphorylated VEGFR-2 expression increased significantly after singleand repeated bevacizumab treatments compared with the control. The 1.0 mg/m L bevacizumab induced significantly higher expressions of VEGFR-2 than the 2.5 mg/m L in single and repeated treatment groups.CONCLUSION： Bevacizumab treatment of HUVECs elevated VEGFR expression in both single and repeated treatments, indicating a mechanism for the reduced efficacy of anti-VEGF therapy in ocular neovascular disorders.
基金This study was supported by the National Research Foundation of Korea(NRF)grant funded by the Korean government(2018R1A6A3A01011692[SHK]from MOE and 2019R1C1C1008999[CH]from MSIT)the National Cancer Center of Korea(NCC-1810102/191050/1911261[BKC]and NCC-2010190[CH]).
文摘The antitumor capabilities of agonistic anti-4-1BB mAbs have made them an attractive target for tumor immunotherapy.However,the adverse side effects associated with agonist antibodies have hindered their clinical development.Here,we aimed to study the immune-related adverse events of repeated doses and long-term use of agonistic anti-4-1BB mAbs.We show that chronic activation of 4-1BB signals induced the accumulation of IFN-γ-producing PD-1^(+)CD8^(+)T cells in the secondary lymphoid organs of tumor-bearing mice by increasing the number of dividing CD8^(+)T cells,which was beneficial for suppressing tumor growth in the early phase of anti-4-1BB induction.However,repeated exposure to anti-4-1BB mAbs led to granuloma development in tumor-draining lymph nodes(TDLNs)of mice due to recruitment and accumulation of macrophages via the CD8^(+)T cell-IFN-γaxis.This was accompanied by excessive lymph node swelling,which impaired the sequential activation of CD8^(+)T cells.Our data provide insights into the immune-related adverse events of long-term agonist 4-1BB antibody dosing,which should be considered during the clinical development of immunomodulating therapy.
基金This study was supported by the National Research Foundation of Korea(NRF)grant funded by the Korean government(2022R1A2C1005463[BKC],2022R1C1C1010078[SHK],and 2022R1C1C1003152[CH]from MSIT)by the National Cancer Center of Korea(NCC)grant funded by the Ministry of Health and Welfare(NCC-2212450[CH]).
文摘4-1BB is an inducible receptor expressed on activated T cells,while its ligand,4-1BBL,is mainly expressed in antigen-presenting cells and macrophages[1,2].To the best of our knowledge,ligandmediated transactivation of 4-1BB is responsible for the survival and immune effector functions of T cells.
基金funding from the European Union’s Horizon 2020 research and innovation program(ERC StG,SIMPHONICS,Project No.101041486)X.Y.acknowledges funding from the Chinese Scholarship Council(Scholarship No.202108270002).All authors acknowledge K.Watanabe and T.Taniguchi from the National Institute of Materials Science(NIMS)for the bulk hBN crystals.
文摘Reading biomolecular signatures and understanding their role in health and disease is one of the greatest scientific challenges in modern biology.Decoding this information is not only foundational for biology but also a cornerstone for next generation molecular diagnostics.This calls for novel methods that can capture the presence and identity of low-abundance compounds at the individual molecule level.Since its inception in the 1980s,nanopore sequencing has become an essential part of the single-molecule sensing toolkit,proving that long,labelfree reads of DNA can be achieved at low cost and high throughput.1 Despite their huge success in genome sequencing,reading the linear sequence of proteins is a considerably more complex task that requires differentiating 20 different amino acids(as opposed to the 4 DNA bases),as well as their modifications,during real-time translocation.Deconvoluting the time-dependent electrical current traces to determine entire amino acid sequences over long reads is as yet an unaccomplished milestone.
