Psoriasis is a common chronic inflammatory skin disease.The diversity and heterogeneity of immune cells in human skin have been studied in recent years,but the spatial distribution of immune cells at the single-cell l...Psoriasis is a common chronic inflammatory skin disease.The diversity and heterogeneity of immune cells in human skin have been studied in recent years,but the spatial distribution of immune cells at the single-cell level in the human psoriatic epidermis and dermis remains unclear.In this study,we mapped psoriatic skin immune cells from paired lesional,perilesional,and nonlesional skin samples using mass cytometry.Phenotypic dendritic cells(DCs)were found in the psoriatic epidermis and dermis.Psoriatic dermal CD1c^(+)CD11b^(+)cDC2s migrated to the epidermis in the perilesional skin during the preinitiation stage.CD1c^(+)CD11b^(+)cDC2s rapidly replaced EpCAM^(+)CD11c^(low) LC cells and initiated inflammation.Simultaneously,CD207^(+)CD11c^(hi) LC and CD5^(+)T cells accumulated in the psoriatic epidermis and orchestrated epidermal inflammation in psoriasis.The immune cell pool in the psoriatic dermis primarily included APCs and T cells.However,unlike that in the dermis,the epidermal immune environment was more significant and coincided with the inflammation occurring during psoriasis.展开更多
Single-cell mass cytometry(SCMC)combines features of traditional flow cytometry(i.e.,fluorescence-activated cell sorting)with mass spectrometry,making it possible to measure several parameters at the single-cell level...Single-cell mass cytometry(SCMC)combines features of traditional flow cytometry(i.e.,fluorescence-activated cell sorting)with mass spectrometry,making it possible to measure several parameters at the single-cell level for a complex analysis of biological regulatory mechanisms.In this study,we optimized SCMC to analyze hemocytes of the Drosophila innate immune system.We used metal-conjugated antibodies(against cell surface antigens H2,H3,H18,L1,L4,and P1,and intracellular antigens 3A5 and L2)and anti-IgM(against cell surface antigen L6)to detect the levels of antigens,while anti-GFP was used to detect crystal cells in the immune-induced samples.We investigated the antigen expression profile of single cells and hemocyte populations in naive states,in immune-induced states,in tumorous mutants bearing a driver mutation in the Drosophila homologue of Janus kinase(hopTum)and carrying a deficiency of the tumor suppressor gene lethal(3)malignant blood neoplasm-1[l(3)mbn1],as well as in stem cell maintenance-defective hdcD84 mutant larvae.Multidimensional analysis enabled the discrimination of the functionally different major hemocyte subsets for lamellocytes,plasmatocytes,and crystal cells,and delineated the unique immunophenotype of Drosophila mutants.We have identified subpopulations of L2^(+)/P1^(+)and L2^(+)/L4^(+)/P1^(+)transitional phenotype cells in the tumorous strains l(3)mbn1 and hopTum,respectively,and a subpopulation of L4^(+)/P1^(+)cells upon immune induction.Our results demonstrated for the first time that SCMC,combined with multidimensional bioinformatic analysis,represents a versatile and powerful tool to deeply analyze the regulation of cell-mediated immunity of Drosophila.展开更多
Susceptibility to pathogens in the elderly is heightened with age,largely because of immunosenescence.As an immune regulatory organ,bone marrow creates immune cells that move to other organs and tis-sues through the b...Susceptibility to pathogens in the elderly is heightened with age,largely because of immunosenescence.As an immune regulatory organ,bone marrow creates immune cells that move to other organs and tis-sues through the blood.Despite the significance of this process of this organ,there is limited research on changes in immune cell generation in the bone marrow and their effects on immunosenescence.In this study,the compositions of immune cells in bone marrow from young(three months)and old(24+months)mice were compared by means of mass cytometry,with further validation obtained through the reanalysis of single-cell RNA sequencing data and cell sorting via flow cytometry.The effects of differential immune cells on immunosenescence in old mice were evaluated using the Clostridium difficile(C.difficile)infection model.Our results showed that aged mice presented with a reduction in bone tra-beculae structure,which was accompanied by a notable increase in polymorphonuclear(PMN)-myeloid-derived suppressor cell(MDSC)abundance.Through bulk-seq and reverse transcription quantitative polymerase chain reaction(RT-qPCR)analysis,we identified differential genes associated with the immune response—specifically,the Th17 cell differentiation pathway.Furthermore,the increase in exported PMN-MDSCs to the large intestine resulted in increased gut permeability and inflammatory damage to the colon following C.difficile infection.After clearing the PMN-MDSCs in old mice using the anti-Gr-1 antibody,the symptoms induced by C.difficile were significantly relieved,as evidenced by an inhibited IL-17 pathway in the colon and reduced gut permeability.In conclusion,aging increases the number of PMN-MDSCs in both the generated bone marrow and the outputted intestine,which con-tributes to susceptibility to C.difficile infection.This study provides a novel target for anti-aging therapy for immunosenescence,which is beneficial for improving immune function in elders.展开更多
Molecular profiling of cell-surface proteins is a powerful strategy for precise cancer diagnosis.While mass cytometry(MC)enables synchronous detection of over 40 cellular parameters,its full potential in disease class...Molecular profiling of cell-surface proteins is a powerful strategy for precise cancer diagnosis.While mass cytometry(MC)enables synchronous detection of over 40 cellular parameters,its full potential in disease classification is challenged by the limited types of recognition probes currently available.In this work,we synthesize a panel of heavy isotopeconjugated aptamers to profile cancer-associated signatures on the surface of hematological malignancy(HM)cells.Based on 15 molecular signatures,we performed cell-surface profiling that allowed the precise classification of 8 HM cell lines.Combined with machine-learning technology,this aptamer-based MC platform also achieved multiclass identification of HM subtypes in clinical sampleswith 100%accuracy in the training cohort and 80%accuracy in the test cohort.Therefore,we report an effective and practical strategy for precise cancer classification at the singlecell level,paving the way for its clinical use in the near future.展开更多
Self-assembling carrier-free nanodrugs are attractive agents because they accumulate at tumor by an enhanced permeability and retention(EPR) effect without introduction of inactive substances,and some nanodrugs can al...Self-assembling carrier-free nanodrugs are attractive agents because they accumulate at tumor by an enhanced permeability and retention(EPR) effect without introduction of inactive substances,and some nanodrugs can alter the immune environment. We synthesized a peptidyl arginine deiminase 4(PAD4) molecular inhibitor, ZD-E-1 M. It could self-assembled into nanodrug ZD-E-1. Using confocal laser scanning microscopy, we observed its cellular colocalization, PAD4 activity and neutrophil extracellular traps(NETs) formation. The populations of immune cells and expression of immune-related proteins were determined by single-cell mass cytometry. ZD-E-1 formed nanoflowers in an acidic environment, whereas it formed nanospheres at pH 7.4. Accumulation of ZD-E-1 at tumor was pHresponsive because of its pH-dependent differences in the size and shape. It could enter the nucleus and bind to PAD4 to prolong the intracellular retention time. In mice, ZD-E-1 inhibited tumor growth and metastasis by inhibiting PAD4 activity and NETs formation. Besides, ZD-E-1 could regulate the ratio of immune cells in LLC tumor-bearing mice. Immunosuppressive proteins like LAG3 were suppressed,while IFN-γ and TNF-a as stimulators of tumor immune response were upregulated. Overall, ZD-E-1 is a self-assembling carrier-free nanodrug that responds to pH, inhibits PAD4 activity, blocks neutrophil extracellular traps formation, and improves the tumor immune microenvironment.展开更多
Although ultra-small nanoclusters(USNCs,<2 nm)have immense application capabilities in biomedicine,the investigation on body-wide organ responses towards USNCs is scant.Here,applying a novel strategy of single-cell...Although ultra-small nanoclusters(USNCs,<2 nm)have immense application capabilities in biomedicine,the investigation on body-wide organ responses towards USNCs is scant.Here,applying a novel strategy of single-cell mass cytometry combined with Nano Genome Atlas of multi-tissues,we systematically evaluate the interactions between the host and calcium phosphate(CaP)USNCs at the organism level.Combining single-cell mass cytometry,and magnetic luminex assay results,we identify dynamic immune responses to CaP USNCs at the single cell resolution.The innate immune is initially activated and followed by adaptive immune activation,as evidenced by dynamic immune cells proportions.Furthermore,using Nano Genome Atlas of multi-tissues,we uncover CaP USNCs induce stronger activation of the immune responses in the cartilage and subchondral bone among the five local tissues while promote metabolic activities in the liver and kidney.Moreover,based on the immunological response profiles,histological evaluation of major organs and local tissue,and a body-wide transcriptomics,we demonstrate that CaP USNCs are not more hazardous than the Food and Drug Administration-approved CaP nanoparticles after 14 days of injection.Our findings provide valuable information on the future clinical applications of USNCs and introduce an innovative strategy to decipher the whole body response to implants.展开更多
Primary or secondary(i.e.,acquired)resistance is a common occurrence in cancer patients and is often associated with high numbers of T regulatory(T_(reg))cells(CD4^(+)CD25^(+)FOXP3^(+)).The approval of ipilimumab and ...Primary or secondary(i.e.,acquired)resistance is a common occurrence in cancer patients and is often associated with high numbers of T regulatory(T_(reg))cells(CD4^(+)CD25^(+)FOXP3^(+)).The approval of ipilimumab and the development of similar pharmacological agents targeting cell surface proteins on T_(reg)cells demonstrates that such intervention may overcome resistance in cancer patients.Hence,the clinical development and subsequent approval of Cytotoxic T Lymphocyte Antigen-4(CTLA-4)targeting agents can serve as a prototype for similar agents.Such new agents aspire to be highly specific and have a reduced toxicity profile while increasing effector T cell function or effector T/T regulatory(T_(eff)/T_(reg))ratio.While clinical development with large molecules has shown the greatest advancement,small molecule inhibitors that target immunomodulation are increasingly entering early clinical investigation.These new small molecule inhibitors often target specific intracellular signaling pathways[e.g.,phosphoinositide-3-kinase delta(PI3K-δ)]that play an important role in regulating the function of T_(reg)cells.This review will summarize the lessons currently applied to develop novel clinical agents that target T_(reg)cells.展开更多
基金supported by grants from the National Natural Science Foundation of China (Nos.81930089,82103709,and 82230104).
文摘Psoriasis is a common chronic inflammatory skin disease.The diversity and heterogeneity of immune cells in human skin have been studied in recent years,but the spatial distribution of immune cells at the single-cell level in the human psoriatic epidermis and dermis remains unclear.In this study,we mapped psoriatic skin immune cells from paired lesional,perilesional,and nonlesional skin samples using mass cytometry.Phenotypic dendritic cells(DCs)were found in the psoriatic epidermis and dermis.Psoriatic dermal CD1c^(+)CD11b^(+)cDC2s migrated to the epidermis in the perilesional skin during the preinitiation stage.CD1c^(+)CD11b^(+)cDC2s rapidly replaced EpCAM^(+)CD11c^(low) LC cells and initiated inflammation.Simultaneously,CD207^(+)CD11c^(hi) LC and CD5^(+)T cells accumulated in the psoriatic epidermis and orchestrated epidermal inflammation in psoriasis.The immune cell pool in the psoriatic dermis primarily included APCs and T cells.However,unlike that in the dermis,the epidermal immune environment was more significant and coincided with the inflammation occurring during psoriasis.
基金the National Research,Development and Innovation Office,Hungary(Grant Nos.GINOP-2.3.2-15-2016-00001,GINOP-2.3.2-15-2016-00030 to LGP,GINOP-2.3.2-15-2016-00035 to E´K,NKFI NN118207 and NKFI K120142 to IA,NKFI 120140 to EK,and OTKA K-131484 to VH)Gabor J.Szebeni was supported by the New National Excellence Program of the Ministry for Innovation and Technology,Hungary(Grant No.UNKP-19-4-SZTE-36)and by the Janos Bolyai Research Scholarship of the Hungarian Academy of Sciences(Grant No.BO/00139/17/8)。
文摘Single-cell mass cytometry(SCMC)combines features of traditional flow cytometry(i.e.,fluorescence-activated cell sorting)with mass spectrometry,making it possible to measure several parameters at the single-cell level for a complex analysis of biological regulatory mechanisms.In this study,we optimized SCMC to analyze hemocytes of the Drosophila innate immune system.We used metal-conjugated antibodies(against cell surface antigens H2,H3,H18,L1,L4,and P1,and intracellular antigens 3A5 and L2)and anti-IgM(against cell surface antigen L6)to detect the levels of antigens,while anti-GFP was used to detect crystal cells in the immune-induced samples.We investigated the antigen expression profile of single cells and hemocyte populations in naive states,in immune-induced states,in tumorous mutants bearing a driver mutation in the Drosophila homologue of Janus kinase(hopTum)and carrying a deficiency of the tumor suppressor gene lethal(3)malignant blood neoplasm-1[l(3)mbn1],as well as in stem cell maintenance-defective hdcD84 mutant larvae.Multidimensional analysis enabled the discrimination of the functionally different major hemocyte subsets for lamellocytes,plasmatocytes,and crystal cells,and delineated the unique immunophenotype of Drosophila mutants.We have identified subpopulations of L2^(+)/P1^(+)and L2^(+)/L4^(+)/P1^(+)transitional phenotype cells in the tumorous strains l(3)mbn1 and hopTum,respectively,and a subpopulation of L4^(+)/P1^(+)cells upon immune induction.Our results demonstrated for the first time that SCMC,combined with multidimensional bioinformatic analysis,represents a versatile and powerful tool to deeply analyze the regulation of cell-mediated immunity of Drosophila.
基金supported by the National Key Research and Development Program of China(2022YFF1100504)the 111 project from the Education Ministry of China(B18053)+2 种基金the National Natural Science Foundation of China(32101938 and 32302758)the China Postdoctoral Science Foundation(2022M723422)the Postdoctoral Fellowship Program of CPSF(GZB20230848).
文摘Susceptibility to pathogens in the elderly is heightened with age,largely because of immunosenescence.As an immune regulatory organ,bone marrow creates immune cells that move to other organs and tis-sues through the blood.Despite the significance of this process of this organ,there is limited research on changes in immune cell generation in the bone marrow and their effects on immunosenescence.In this study,the compositions of immune cells in bone marrow from young(three months)and old(24+months)mice were compared by means of mass cytometry,with further validation obtained through the reanalysis of single-cell RNA sequencing data and cell sorting via flow cytometry.The effects of differential immune cells on immunosenescence in old mice were evaluated using the Clostridium difficile(C.difficile)infection model.Our results showed that aged mice presented with a reduction in bone tra-beculae structure,which was accompanied by a notable increase in polymorphonuclear(PMN)-myeloid-derived suppressor cell(MDSC)abundance.Through bulk-seq and reverse transcription quantitative polymerase chain reaction(RT-qPCR)analysis,we identified differential genes associated with the immune response—specifically,the Th17 cell differentiation pathway.Furthermore,the increase in exported PMN-MDSCs to the large intestine resulted in increased gut permeability and inflammatory damage to the colon following C.difficile infection.After clearing the PMN-MDSCs in old mice using the anti-Gr-1 antibody,the symptoms induced by C.difficile were significantly relieved,as evidenced by an inhibited IL-17 pathway in the colon and reduced gut permeability.In conclusion,aging increases the number of PMN-MDSCs in both the generated bone marrow and the outputted intestine,which con-tributes to susceptibility to C.difficile infection.This study provides a novel target for anti-aging therapy for immunosenescence,which is beneficial for improving immune function in elders.
基金the National Key Research Program(grant nos.2021YFA0910101,2018YFC1602900,and 2019YFA0905800)the National Natural Science Foundation of China(NSFC+1 种基金grant nos.21922404,22174039,22107027,and 21827811)the Science and Technology Project of Hunan Province(grant nos.2022JJ10005,2021RC4022,2019SK2201,2018RS3035,and 2017XK2103).
文摘Molecular profiling of cell-surface proteins is a powerful strategy for precise cancer diagnosis.While mass cytometry(MC)enables synchronous detection of over 40 cellular parameters,its full potential in disease classification is challenged by the limited types of recognition probes currently available.In this work,we synthesize a panel of heavy isotopeconjugated aptamers to profile cancer-associated signatures on the surface of hematological malignancy(HM)cells.Based on 15 molecular signatures,we performed cell-surface profiling that allowed the precise classification of 8 HM cell lines.Combined with machine-learning technology,this aptamer-based MC platform also achieved multiclass identification of HM subtypes in clinical sampleswith 100%accuracy in the training cohort and 80%accuracy in the test cohort.Therefore,we report an effective and practical strategy for precise cancer classification at the singlecell level,paving the way for its clinical use in the near future.
基金supported by the Beijing Municipal Colleges and Universities High Level Talents Introduction and Cultivate Project-Beijing Great Wall Scholar Program (CIT&TCD 20180332,China)。
文摘Self-assembling carrier-free nanodrugs are attractive agents because they accumulate at tumor by an enhanced permeability and retention(EPR) effect without introduction of inactive substances,and some nanodrugs can alter the immune environment. We synthesized a peptidyl arginine deiminase 4(PAD4) molecular inhibitor, ZD-E-1 M. It could self-assembled into nanodrug ZD-E-1. Using confocal laser scanning microscopy, we observed its cellular colocalization, PAD4 activity and neutrophil extracellular traps(NETs) formation. The populations of immune cells and expression of immune-related proteins were determined by single-cell mass cytometry. ZD-E-1 formed nanoflowers in an acidic environment, whereas it formed nanospheres at pH 7.4. Accumulation of ZD-E-1 at tumor was pHresponsive because of its pH-dependent differences in the size and shape. It could enter the nucleus and bind to PAD4 to prolong the intracellular retention time. In mice, ZD-E-1 inhibited tumor growth and metastasis by inhibiting PAD4 activity and NETs formation. Besides, ZD-E-1 could regulate the ratio of immune cells in LLC tumor-bearing mice. Immunosuppressive proteins like LAG3 were suppressed,while IFN-γ and TNF-a as stimulators of tumor immune response were upregulated. Overall, ZD-E-1 is a self-assembling carrier-free nanodrug that responds to pH, inhibits PAD4 activity, blocks neutrophil extracellular traps formation, and improves the tumor immune microenvironment.
基金the National Key Research and Development Program of China(2018YFC1105100)NSFC grants(T2121004,81972099,82072463,81871764)+1 种基金Zhejiang Provincial Natural Science Foundation of China(LZ22H060002,LR20H060001)Fundamental Research Funds for the Central Universities.
文摘Although ultra-small nanoclusters(USNCs,<2 nm)have immense application capabilities in biomedicine,the investigation on body-wide organ responses towards USNCs is scant.Here,applying a novel strategy of single-cell mass cytometry combined with Nano Genome Atlas of multi-tissues,we systematically evaluate the interactions between the host and calcium phosphate(CaP)USNCs at the organism level.Combining single-cell mass cytometry,and magnetic luminex assay results,we identify dynamic immune responses to CaP USNCs at the single cell resolution.The innate immune is initially activated and followed by adaptive immune activation,as evidenced by dynamic immune cells proportions.Furthermore,using Nano Genome Atlas of multi-tissues,we uncover CaP USNCs induce stronger activation of the immune responses in the cartilage and subchondral bone among the five local tissues while promote metabolic activities in the liver and kidney.Moreover,based on the immunological response profiles,histological evaluation of major organs and local tissue,and a body-wide transcriptomics,we demonstrate that CaP USNCs are not more hazardous than the Food and Drug Administration-approved CaP nanoparticles after 14 days of injection.Our findings provide valuable information on the future clinical applications of USNCs and introduce an innovative strategy to decipher the whole body response to implants.
文摘Primary or secondary(i.e.,acquired)resistance is a common occurrence in cancer patients and is often associated with high numbers of T regulatory(T_(reg))cells(CD4^(+)CD25^(+)FOXP3^(+)).The approval of ipilimumab and the development of similar pharmacological agents targeting cell surface proteins on T_(reg)cells demonstrates that such intervention may overcome resistance in cancer patients.Hence,the clinical development and subsequent approval of Cytotoxic T Lymphocyte Antigen-4(CTLA-4)targeting agents can serve as a prototype for similar agents.Such new agents aspire to be highly specific and have a reduced toxicity profile while increasing effector T cell function or effector T/T regulatory(T_(eff)/T_(reg))ratio.While clinical development with large molecules has shown the greatest advancement,small molecule inhibitors that target immunomodulation are increasingly entering early clinical investigation.These new small molecule inhibitors often target specific intracellular signaling pathways[e.g.,phosphoinositide-3-kinase delta(PI3K-δ)]that play an important role in regulating the function of T_(reg)cells.This review will summarize the lessons currently applied to develop novel clinical agents that target T_(reg)cells.
