Ultraviolet blood irradiation has been used as a physical therapy to treat many nonspeci¯c diseases in clinics;however,the underlying mechanisms remain largely unclear.Neutrophils,the first line of host defense,p...Ultraviolet blood irradiation has been used as a physical therapy to treat many nonspeci¯c diseases in clinics;however,the underlying mechanisms remain largely unclear.Neutrophils,the first line of host defense,play a crucial role in a variety of in°ammatory responses.In the present work,we investigated the effects of ultraviolet light A(UVA)on the immune functions of human neutrophils at the single-cell level by using an inverted°uorescence microscope.N-Formylmethionyl-leucyl-phenylalanine(FMLP),a classic physiological chemotactic peptide,was used to induce a series of immune responses in neutrophils in vitro.FMLP-induced calcium mobilization,migration,and phagocytosis in human neutrophils was significantly blocked after treatment with 365 nm UVA irradiation,demonstrating the immunosuppressive effects of UVA irradiation on neutrophils.Similar responses were also observed when the cells were pretreated with H2O2,a type of reactive oxygen species(ROS).Furthermore,UVA irradiation resulted in an increase in NAD(P)H,a member of host oxidative stress in cells.Taken together,our data indicate that UVA irradiation results in immunosuppression associated with the production of ROS in human neutrophils.展开更多
In order to clarify, the mechanism of inhibition of human neutrophil peptide-1 （ HNP-1 ） on hu- man immunodeficiency vires type 1 （HIV-1 ）, CD4^ ＋ cells were used as the target cells for acute infection with HIV-...In order to clarify, the mechanism of inhibition of human neutrophil peptide-1 （ HNP-1 ） on hu- man immunodeficiency vires type 1 （HIV-1 ）, CD4^ ＋ cells were used as the target cells for acute infection with HIV-1, and experiments were peffomed separately with the interaction of different concentrations of HNP-1 with free vires particles, un-infected and infected CD4^＋ cells. The activity of reverse transcriptase （RT） in the supematant of cell cultures of different lots of experiments were then assayed accordingly, and the toxicity effect on human lymphocytic cells MT4 was measured by MTT assay. The experimental results showed that pre-incubation of HNP-1 with the concentrated stock of vires could block the binding of vires to target cells with EC50 of 2.49 μg/ml, while pre-treatment of CD4^＋ cells with HNP- 1 prior to inoculation could reduce the ability of cells to bind vires with EC50 of 20.7 μg/ml. In addition, When culturing the infected CD4^＋ cells in the continuous presence of various concentrations of HNP-1 added immediately after infection, HNP-1 exhibited modest inhibitory effect on viral replication with reduced RT activities in comparison with those of the control group （ P 〈 0.05 at 100 μg/ml of the highest concentration） . No cytotoxieity effect of HNP-1 was observed as demonstrated by MTT assay. These results indicate that HNP-1 exerts anti-HIV activity by at least two levels： direct inactivation of vires particles and effect on the ability of target cells to bind with viruses. The evaluation of two parameters, inhibitoty effect and the cytotoxicity renders HNP-1 an available candidate for anti-HIV therapeutic agent.展开更多
Background:Human neutrophil lipocalin(HNL)has been used extensively to differentiate acute bacterial infection from febrile diseases as a biomarker to reflect the activation of the neutrophil.The serum HNL levels in t...Background:Human neutrophil lipocalin(HNL)has been used extensively to differentiate acute bacterial infection from febrile diseases as a biomarker to reflect the activation of the neutrophil.The serum HNL levels in the adult-onset Still’s disease(AOSD)patients with and without infection,as well as the healthy controls(HCs),were analyzed statistically in this study to evaluate the value of HNL for the diagnosis of AOSD.Methods:A total of 129 AOSD patients were enrolled,from whom blood samples were drawn and the AOSD diagnosis was confirmed through the review of the medical records,where the systemic score,demographic characteristics,clinical manifestations,and laboratory parameters were also collected for the patients;in addition,a total of 40 HCs were recruited among the blood donors from the healthcare center with the relevant information collected.The HNL test was done for the blood samples with the enzyme-linked immunosorbent assay and the analyses were done for the correlations of HNL with clinical manifestations and diagnostic effectiveness.Results:The serum HNL increased significantly in the patients with only AOSD as compared with that in the HCs(139.76±8.99 ng/mL vs.55.92±6.12 ng/mL;P<0.001).The serum HNL level was correlated with the white blood cell(WBC)count(r=0.335,P<0.001),neutrophil count(r=0.334,P<0.001),erythrocyte sedimentation rate(r=0.241,P=0.022),C-reactive protein(r=0.442,P<0.0001),and systemic score(r=0.343,P<0.0001)in the AOSD patients significantly.Patients with fever,leukocytosis≥15,000/mm^(3),and myalgia in the HNL-positive group were observed relatively more than those in the HNL-negative group(P=0.009,P=0.023,and P=0.007,respectively).HNL was a more sensitive indicator than ferritin and C-reactive protein(CRP)to differentiate the AOSD patients with bacterial infection from AOSD-only patients,and the Youden index was 0.6 for HNL and 0.29 for CRP.Conclusion:Serum HNL can be used as a biomarker for the diagnosis of the AOSD,and HNL is also observed to be associated with the disease activity.展开更多
Aim:Although treatment of lung cancer,one of the deadliest diseases worldwide,with immune checkpoint inhibitors(ICIs)has shown promising outcomes,these survival outcomes are only observed in a relatively small subset ...Aim:Although treatment of lung cancer,one of the deadliest diseases worldwide,with immune checkpoint inhibitors(ICIs)has shown promising outcomes,these survival outcomes are only observed in a relatively small subset of lung cancer patients.In a previous study,we elucidated that the presence of human neutrophil peptide(HNP)1,2 and 3 in non-small cell lung cancerous(NSCLC)biopsies is associated with a clinical response towards treatment with PD-1/PD-L1 immune checkpoint inhibitors.Furthermore,HNP1 has shown in vitro an immune-activated function towards lung cancer cells,but the specific role of HNP1 in(lung)cancer is still unknown.The aim of this study was to provide a better understanding of HNP1 in an NSCLC microenvironment.Methods:To gain better insights into the role of HNP1 on cancer growth and to unravel immune responses,in vitro(SILAC-labelled)A549/PBMC(from three healthy donors)cocultures were set up and treated/not treated with HNP1.After 5 days,both secretome and cellular analysis using mass spectrometry were performed on these cocultures.After protein identification in all different tested conditions,pathway analyses(MetaCoreTM)were performed to investigate the biological significance of HNP1 stimulation on the A549/PBMC cocultures.The proteomic outcomes were confirmed by multiplex ELISA for a proinflammatory cytokine panel(TNF-α,IL-1β,IL-2,IL-4,IL-6,IL-12p70 and IL-18).Results:A number of biological pathways and process networks were observed to be upregulated after treatment of the coculture with HNP1.HNP1 stimulation leads to an increase in pathways and proteins stimulating chemotaxis(including plasmin signaling,leucocyte recruitment,CCL2 and CXCL8 expression),proinflammatory cytokine secretion(including IL-1β,IL-6 and TNF-α),dendritic cell(DC)maturation,phagocytosis and antigen presentation,leading to a more efficient adaptive anti-tumoral immunity.Conclusion:These results enhance our understanding of the role of HNP1 in the tumor microenvironment and suggest that HNP1 may be able to induce tumor necrosis by inducing prostimulatory immune responses.展开更多
The primary cause of mortality at 5 years following a cardiac transplantation is the development of atherosclerosis, termed coronary allograft vasculopathy （CAV）. This pathology is characterized by diffused intimal ...The primary cause of mortality at 5 years following a cardiac transplantation is the development of atherosclerosis, termed coronary allograft vasculopathy （CAV）. This pathology is characterized by diffused intimal hyperplasia and emanates from coronary arterial injuries caused by immune inflammatory cells. Neutrophils play an important role in this inflammatory process; however, their potential participation in the pathogenesis of CAV is poorly understood. Despite their essential contribution to the prevention of graft rejection, immunosuppressive drugs could have detrimental effects owing to their pro-inflammatory activities. Thus, we investigated the impact of different immunosuppressive drugs on the inflammatory response of neutrophils isolated from the blood of healthy volunteers. Under basal conditions, mammalian target of rapamycin （mTOR） inhibitors （sirolimus and everolimus） had the most potent anti-inflammatory effect, decreasing both IL-8 release （≈-80%） and vascular endothelial growth factor （VEGF） release （≈-65%） and preserving the release of the anti-inflammatory cytokine interleukin-1 receptor antagonist （IL-1RA）. In TNF-α-treated neutrophils, pre-incubation with everolimus provided the most potent effect, simultaneously reducing the release of both VEGF and IL-8 while doubling the release of IL-1RA. This latter effect of everolimus was maintained even when administered in combination with other immunosuppressive drugs. Sirolimus and everolimus decreased the tumor necrosis factor （TNF）-α-induced adhesion of neutrophils to human endothelial cells and human extracellular matrix. This effect was largely dependent on the ability of these compounds to alter P2-integrin/CD18 activation. Our results suggest a potential mechanism for the beneficial effect of everolimus in the prevention of CAV in heart transplant recipients.展开更多
Selectins are carbohydrate-binding cell adhesion molecules that play a major role in the initiation of inflammatory responses. Accumulaed evidence has suggested that heparin＇s anti-inflammatory effects are mainly med...Selectins are carbohydrate-binding cell adhesion molecules that play a major role in the initiation of inflammatory responses. Accumulaed evidence has suggested that heparin＇s anti-inflammatory effects are mainly mediated by blocking L-or P-selectin-initiated cell adhesion. Recently, we have reported that periodate-oxidized, borohydridereduced heparin （RO-heparln） can inhibit P-selectin-mediated acute inflammation. Here we further examined the effect of RO-heparin on the adhesion of L-selectin-mediated leukocytes to vascular endothelium under flow conditions in vivo and in vitro. The results show that RO-heparin with a low anticoagulant activity can effectively reduce leucocyte roiling on thioglycoUate-induced rat mesenterlc venules and L-selectin-metadiated neutrophil roiling on TNF-α-induced human umbilical vein endothelial cells（HUVECs） under flow conditions. Our findings suggest that the effect of RO-heparin on inflammatory responses is mainly a result of its inhibiting the interaction between P- or L-selectin and its ligands. The findings also suggest that RO-heparin may be useful in preventing inflammation diseases.展开更多
基金This work was supported by the National Natural Science Foundation of China(No.11204142),the National Basic Research Program of China (No.2013CB328702)International S&T Cooperation Program of China (2011DFA52870)+1 种基金the 111 Project(No.B07013),the National Science Fund for Talent Training in Basic Sciences(No.J1103208),the PCSIRT (IRT0149)the Social Development Fund of Ge'ermu Science and Technology Bureau and the State Key Laboratory of Medicinal Chemical Biology.
文摘Ultraviolet blood irradiation has been used as a physical therapy to treat many nonspeci¯c diseases in clinics;however,the underlying mechanisms remain largely unclear.Neutrophils,the first line of host defense,play a crucial role in a variety of in°ammatory responses.In the present work,we investigated the effects of ultraviolet light A(UVA)on the immune functions of human neutrophils at the single-cell level by using an inverted°uorescence microscope.N-Formylmethionyl-leucyl-phenylalanine(FMLP),a classic physiological chemotactic peptide,was used to induce a series of immune responses in neutrophils in vitro.FMLP-induced calcium mobilization,migration,and phagocytosis in human neutrophils was significantly blocked after treatment with 365 nm UVA irradiation,demonstrating the immunosuppressive effects of UVA irradiation on neutrophils.Similar responses were also observed when the cells were pretreated with H2O2,a type of reactive oxygen species(ROS).Furthermore,UVA irradiation resulted in an increase in NAD(P)H,a member of host oxidative stress in cells.Taken together,our data indicate that UVA irradiation results in immunosuppression associated with the production of ROS in human neutrophils.
文摘In order to clarify, the mechanism of inhibition of human neutrophil peptide-1 （ HNP-1 ） on hu- man immunodeficiency vires type 1 （HIV-1 ）, CD4^ ＋ cells were used as the target cells for acute infection with HIV-1, and experiments were peffomed separately with the interaction of different concentrations of HNP-1 with free vires particles, un-infected and infected CD4^＋ cells. The activity of reverse transcriptase （RT） in the supematant of cell cultures of different lots of experiments were then assayed accordingly, and the toxicity effect on human lymphocytic cells MT4 was measured by MTT assay. The experimental results showed that pre-incubation of HNP-1 with the concentrated stock of vires could block the binding of vires to target cells with EC50 of 2.49 μg/ml, while pre-treatment of CD4^＋ cells with HNP- 1 prior to inoculation could reduce the ability of cells to bind vires with EC50 of 20.7 μg/ml. In addition, When culturing the infected CD4^＋ cells in the continuous presence of various concentrations of HNP-1 added immediately after infection, HNP-1 exhibited modest inhibitory effect on viral replication with reduced RT activities in comparison with those of the control group （ P 〈 0.05 at 100 μg/ml of the highest concentration） . No cytotoxieity effect of HNP-1 was observed as demonstrated by MTT assay. These results indicate that HNP-1 exerts anti-HIV activity by at least two levels： direct inactivation of vires particles and effect on the ability of target cells to bind with viruses. The evaluation of two parameters, inhibitoty effect and the cytotoxicity renders HNP-1 an available candidate for anti-HIV therapeutic agent.
基金supported by grants from the National Natural Science Foundation of China(Nos.81871281,81801618,82171773,81971523,U1903210)China International Medical Foundation(No.Z-2018-40-2101)the Clinical Medicine Plus X-Young Scholars Project of Peking University(No.PKU2021LCXQ008)by the Fundamental Research Funds for the Central Universities.
文摘Background:Human neutrophil lipocalin(HNL)has been used extensively to differentiate acute bacterial infection from febrile diseases as a biomarker to reflect the activation of the neutrophil.The serum HNL levels in the adult-onset Still’s disease(AOSD)patients with and without infection,as well as the healthy controls(HCs),were analyzed statistically in this study to evaluate the value of HNL for the diagnosis of AOSD.Methods:A total of 129 AOSD patients were enrolled,from whom blood samples were drawn and the AOSD diagnosis was confirmed through the review of the medical records,where the systemic score,demographic characteristics,clinical manifestations,and laboratory parameters were also collected for the patients;in addition,a total of 40 HCs were recruited among the blood donors from the healthcare center with the relevant information collected.The HNL test was done for the blood samples with the enzyme-linked immunosorbent assay and the analyses were done for the correlations of HNL with clinical manifestations and diagnostic effectiveness.Results:The serum HNL increased significantly in the patients with only AOSD as compared with that in the HCs(139.76±8.99 ng/mL vs.55.92±6.12 ng/mL;P<0.001).The serum HNL level was correlated with the white blood cell(WBC)count(r=0.335,P<0.001),neutrophil count(r=0.334,P<0.001),erythrocyte sedimentation rate(r=0.241,P=0.022),C-reactive protein(r=0.442,P<0.0001),and systemic score(r=0.343,P<0.0001)in the AOSD patients significantly.Patients with fever,leukocytosis≥15,000/mm^(3),and myalgia in the HNL-positive group were observed relatively more than those in the HNL-negative group(P=0.009,P=0.023,and P=0.007,respectively).HNL was a more sensitive indicator than ferritin and C-reactive protein(CRP)to differentiate the AOSD patients with bacterial infection from AOSD-only patients,and the Youden index was 0.6 for HNL and 0.29 for CRP.Conclusion:Serum HNL can be used as a biomarker for the diagnosis of the AOSD,and HNL is also observed to be associated with the disease activity.
文摘Aim:Although treatment of lung cancer,one of the deadliest diseases worldwide,with immune checkpoint inhibitors(ICIs)has shown promising outcomes,these survival outcomes are only observed in a relatively small subset of lung cancer patients.In a previous study,we elucidated that the presence of human neutrophil peptide(HNP)1,2 and 3 in non-small cell lung cancerous(NSCLC)biopsies is associated with a clinical response towards treatment with PD-1/PD-L1 immune checkpoint inhibitors.Furthermore,HNP1 has shown in vitro an immune-activated function towards lung cancer cells,but the specific role of HNP1 in(lung)cancer is still unknown.The aim of this study was to provide a better understanding of HNP1 in an NSCLC microenvironment.Methods:To gain better insights into the role of HNP1 on cancer growth and to unravel immune responses,in vitro(SILAC-labelled)A549/PBMC(from three healthy donors)cocultures were set up and treated/not treated with HNP1.After 5 days,both secretome and cellular analysis using mass spectrometry were performed on these cocultures.After protein identification in all different tested conditions,pathway analyses(MetaCoreTM)were performed to investigate the biological significance of HNP1 stimulation on the A549/PBMC cocultures.The proteomic outcomes were confirmed by multiplex ELISA for a proinflammatory cytokine panel(TNF-α,IL-1β,IL-2,IL-4,IL-6,IL-12p70 and IL-18).Results:A number of biological pathways and process networks were observed to be upregulated after treatment of the coculture with HNP1.HNP1 stimulation leads to an increase in pathways and proteins stimulating chemotaxis(including plasmin signaling,leucocyte recruitment,CCL2 and CXCL8 expression),proinflammatory cytokine secretion(including IL-1β,IL-6 and TNF-α),dendritic cell(DC)maturation,phagocytosis and antigen presentation,leading to a more efficient adaptive anti-tumoral immunity.Conclusion:These results enhance our understanding of the role of HNP1 in the tumor microenvironment and suggest that HNP1 may be able to induce tumor necrosis by inducing prostimulatory immune responses.
文摘The primary cause of mortality at 5 years following a cardiac transplantation is the development of atherosclerosis, termed coronary allograft vasculopathy （CAV）. This pathology is characterized by diffused intimal hyperplasia and emanates from coronary arterial injuries caused by immune inflammatory cells. Neutrophils play an important role in this inflammatory process; however, their potential participation in the pathogenesis of CAV is poorly understood. Despite their essential contribution to the prevention of graft rejection, immunosuppressive drugs could have detrimental effects owing to their pro-inflammatory activities. Thus, we investigated the impact of different immunosuppressive drugs on the inflammatory response of neutrophils isolated from the blood of healthy volunteers. Under basal conditions, mammalian target of rapamycin （mTOR） inhibitors （sirolimus and everolimus） had the most potent anti-inflammatory effect, decreasing both IL-8 release （≈-80%） and vascular endothelial growth factor （VEGF） release （≈-65%） and preserving the release of the anti-inflammatory cytokine interleukin-1 receptor antagonist （IL-1RA）. In TNF-α-treated neutrophils, pre-incubation with everolimus provided the most potent effect, simultaneously reducing the release of both VEGF and IL-8 while doubling the release of IL-1RA. This latter effect of everolimus was maintained even when administered in combination with other immunosuppressive drugs. Sirolimus and everolimus decreased the tumor necrosis factor （TNF）-α-induced adhesion of neutrophils to human endothelial cells and human extracellular matrix. This effect was largely dependent on the ability of these compounds to alter P2-integrin/CD18 activation. Our results suggest a potential mechanism for the beneficial effect of everolimus in the prevention of CAV in heart transplant recipients.
文摘Selectins are carbohydrate-binding cell adhesion molecules that play a major role in the initiation of inflammatory responses. Accumulaed evidence has suggested that heparin＇s anti-inflammatory effects are mainly mediated by blocking L-or P-selectin-initiated cell adhesion. Recently, we have reported that periodate-oxidized, borohydridereduced heparin （RO-heparln） can inhibit P-selectin-mediated acute inflammation. Here we further examined the effect of RO-heparin on the adhesion of L-selectin-mediated leukocytes to vascular endothelium under flow conditions in vivo and in vitro. The results show that RO-heparin with a low anticoagulant activity can effectively reduce leucocyte roiling on thioglycoUate-induced rat mesenterlc venules and L-selectin-metadiated neutrophil roiling on TNF-α-induced human umbilical vein endothelial cells（HUVECs） under flow conditions. Our findings suggest that the effect of RO-heparin on inflammatory responses is mainly a result of its inhibiting the interaction between P- or L-selectin and its ligands. The findings also suggest that RO-heparin may be useful in preventing inflammation diseases.
