Objective and Impact Statement.We propose a rapid and accurate blood cell identification method exploiting deep learning and label-free refractive index(RI)tomography.Our computational approach that fully utilizes tom...Objective and Impact Statement.We propose a rapid and accurate blood cell identification method exploiting deep learning and label-free refractive index(RI)tomography.Our computational approach that fully utilizes tomographic information of bone marrow(BM)white blood cell(WBC)enables us to not only classify the blood cells with deep learning but also quantitatively study their morphological and biochemical properties for hematology research.Introduction.Conventional methods for examining blood cells,such as blood smear analysis by medical professionals and fluorescence-activated cell sorting,require significant time,costs,and domain knowledge that could affect test results.While label-free imaging techniques that use a specimen’s intrinsic contrast(e.g.,multiphoton and Raman microscopy)have been used to characterize blood cells,their imaging procedures and instrumentations are relatively time-consuming and complex.Methods.The RI tomograms of the BM WBCs are acquired via Mach-Zehnder interferometer-based tomographic microscope and classified by a 3D convolutional neural network.We test our deep learning classifier for the four types of bone marrow WBC collected from healthy donors(n=10):monocyte,myelocyte,B lymphocyte,and T lymphocyte.The quantitative parameters of WBC are directly obtained from the tomograms.Results.Our results show>99%accuracy for the binary classification of myeloids and lymphoids and>96%accuracy for the four-type classification of B and T lymphocytes,monocyte,and myelocytes.The feature learning capability of our approach is visualized via an unsupervised dimension reduction technique.Conclusion.We envision that the proposed cell classification framework can be easily integrated into existing blood cell investigation workflows,providing cost-effective and rapid diagnosis for hematologic malignancy.展开更多
Dendritic cell(DC)-based vaccines continue to be considered an attractive tool for cancer immunotherapy.DCs require an additional signal from the environment or other immune cells to polarize the development of immune...Dendritic cell(DC)-based vaccines continue to be considered an attractive tool for cancer immunotherapy.DCs require an additional signal from the environment or other immune cells to polarize the development of immune responses toward T helper 1(Th1)or Th2 responses.DCs play a role in natural killer(NK)cell activation,and NK cells are also able to activate and induce the maturation of DCs.We investigated the types of NK cells that can induce the maturation and enhanced function of DCs and the conditions under which these interactions occur.DCs that were activated by resting NK cells in the presence of inflammatory cytokines exhibited increased expression of several costimulatory molecules and an enhanced ability to produce IL-12p70.NK cell-stimulated DCs potently induced Th1 polarization and exhibited the ability to generate tumor antigen-specific cytotoxic T lymphocyte responses.Our data demonstrate that functional DCs can be generated by coculturing immature DCs with freshly isolated resting NK cells in the presence of Toll-like receptor agonists and proinflammatory cytokines and that the resulting DCs effectively present antigens to induce tumor-specific T-cell responses,which suggests that these cells may be useful for cancer immunotherapy.展开更多
The use of natural killer(NK)cells is a promising and safe immunotherapeutic approach in the field of cancer immunotherapy.However,combination treatments are required to enhance the effector functions and therapeutic ...The use of natural killer(NK)cells is a promising and safe immunotherapeutic approach in the field of cancer immunotherapy.However,combination treatments are required to enhance the effector functions and therapeutic efficacy of NK cells.In this study,we investigated the potential of daratumumab(Dara),bortezomib,and dexamethasone(Dvd)to augment the antitumor effects of NK cells in a multiple myeloma(MM)xenograft mouse model.NK cells were expanded and activated using the K562-OX40 ligand and membrane-bound IL-18 and IL-21 in the presence of IL-2 and IL-15 from peripheral blood mononuclear cells from MM patients.A human MM xenograft model was established using human RPMI8226-RFP-FLuc cells in NOD/SCID IL-2Rγnull(NSG)mice.Tumor-bearing mice were divided into six treatment groups:no treatment,expanded NK cells(eNKs),Dara,Dara+eNKs,Dvd,and Dvd+eNKs.Dvd treatment strongly enhanced the cytotoxicity of eNKs by upregulating expression of NK cell activation ligands,downregulating expression of NK cell inhibitory ligands,and promoting antibody-dependent cellular cytotoxicity.The combination of eNKs with Dvd significantly prolonged mouse survival and reduced the tumor burden and serum M-protein level.Furthermore,Dvd pretreatment significantly increased eNK persistence and homing to MM sites.Our findings suggest that Dvd treatment potentiates the antimyeloma effects of NK cells expanded and activated ex vivo by modulating immune responses in MM-bearing mice.展开更多
基金supported by KAIST UP program,BK21+program,Tomocube,National Research Foundation of Korea(2015R1A3A2066550)Institute of Information&Communications Technology Planning&Evaluation(IITP)grant funded by the Korea government(MSIT)(2021-0-00745).
文摘Objective and Impact Statement.We propose a rapid and accurate blood cell identification method exploiting deep learning and label-free refractive index(RI)tomography.Our computational approach that fully utilizes tomographic information of bone marrow(BM)white blood cell(WBC)enables us to not only classify the blood cells with deep learning but also quantitatively study their morphological and biochemical properties for hematology research.Introduction.Conventional methods for examining blood cells,such as blood smear analysis by medical professionals and fluorescence-activated cell sorting,require significant time,costs,and domain knowledge that could affect test results.While label-free imaging techniques that use a specimen’s intrinsic contrast(e.g.,multiphoton and Raman microscopy)have been used to characterize blood cells,their imaging procedures and instrumentations are relatively time-consuming and complex.Methods.The RI tomograms of the BM WBCs are acquired via Mach-Zehnder interferometer-based tomographic microscope and classified by a 3D convolutional neural network.We test our deep learning classifier for the four types of bone marrow WBC collected from healthy donors(n=10):monocyte,myelocyte,B lymphocyte,and T lymphocyte.The quantitative parameters of WBC are directly obtained from the tomograms.Results.Our results show>99%accuracy for the binary classification of myeloids and lymphoids and>96%accuracy for the four-type classification of B and T lymphocytes,monocyte,and myelocytes.The feature learning capability of our approach is visualized via an unsupervised dimension reduction technique.Conclusion.We envision that the proposed cell classification framework can be easily integrated into existing blood cell investigation workflows,providing cost-effective and rapid diagnosis for hematologic malignancy.
基金grant no.RTI05-01-01 from the Regional Technology Innovation Program of the Ministry of Commerce,Industry and Energy,Republic of Korea.
文摘Dendritic cell(DC)-based vaccines continue to be considered an attractive tool for cancer immunotherapy.DCs require an additional signal from the environment or other immune cells to polarize the development of immune responses toward T helper 1(Th1)or Th2 responses.DCs play a role in natural killer(NK)cell activation,and NK cells are also able to activate and induce the maturation of DCs.We investigated the types of NK cells that can induce the maturation and enhanced function of DCs and the conditions under which these interactions occur.DCs that were activated by resting NK cells in the presence of inflammatory cytokines exhibited increased expression of several costimulatory molecules and an enhanced ability to produce IL-12p70.NK cell-stimulated DCs potently induced Th1 polarization and exhibited the ability to generate tumor antigen-specific cytotoxic T lymphocyte responses.Our data demonstrate that functional DCs can be generated by coculturing immature DCs with freshly isolated resting NK cells in the presence of Toll-like receptor agonists and proinflammatory cytokines and that the resulting DCs effectively present antigens to induce tumor-specific T-cell responses,which suggests that these cells may be useful for cancer immunotherapy.
基金supported by grants from the Basic Science Research Program through the National Research Foundation of Korea(NRF)funded by the Ministry of Education,Science and Technology(2018R1A2B6006200,2018R1A5A2024181,and 2020R1A2C2010098).
文摘The use of natural killer(NK)cells is a promising and safe immunotherapeutic approach in the field of cancer immunotherapy.However,combination treatments are required to enhance the effector functions and therapeutic efficacy of NK cells.In this study,we investigated the potential of daratumumab(Dara),bortezomib,and dexamethasone(Dvd)to augment the antitumor effects of NK cells in a multiple myeloma(MM)xenograft mouse model.NK cells were expanded and activated using the K562-OX40 ligand and membrane-bound IL-18 and IL-21 in the presence of IL-2 and IL-15 from peripheral blood mononuclear cells from MM patients.A human MM xenograft model was established using human RPMI8226-RFP-FLuc cells in NOD/SCID IL-2Rγnull(NSG)mice.Tumor-bearing mice were divided into six treatment groups:no treatment,expanded NK cells(eNKs),Dara,Dara+eNKs,Dvd,and Dvd+eNKs.Dvd treatment strongly enhanced the cytotoxicity of eNKs by upregulating expression of NK cell activation ligands,downregulating expression of NK cell inhibitory ligands,and promoting antibody-dependent cellular cytotoxicity.The combination of eNKs with Dvd significantly prolonged mouse survival and reduced the tumor burden and serum M-protein level.Furthermore,Dvd pretreatment significantly increased eNK persistence and homing to MM sites.Our findings suggest that Dvd treatment potentiates the antimyeloma effects of NK cells expanded and activated ex vivo by modulating immune responses in MM-bearing mice.
