Objective:Neutrophils are one of the most predominant infiltrating leukocytes in lung cancer tissues and are associated with lung cancer progression.How neutrophils promote lung cancer progression,however,has not been...Objective:Neutrophils are one of the most predominant infiltrating leukocytes in lung cancer tissues and are associated with lung cancer progression.How neutrophils promote lung cancer progression,however,has not been established.Methods:Kaplan–Meier plotter online analysis and tissue immunohistochemistry were used to determine the relationship between neutrophils and overall survival in lung cancer patients.The effect of neutrophils on lung cancer was determined using the Transwell migration assay,a proliferation assay,and a murine tumor model.Gene knockdown was used to determine poly ADPribose polymerase(PARP)-1 function in lung cancer-educated neutrophils.Western blot analysis and gelatin zymography were used to demonstrate the correlation between PARP-1 and matrix metallopeptidase 9(MMP-9).Immunoprecipitation coupled to mass spectrometry(IP/MS)was used to identify the proteins interacting with PARP-1.Co-immunoprecipitation(Co-IP)was used to confirm that PARP-1 interacts with arachidonate 5-lipooxygenase(ALOX5).Neutrophil PARP-1 blockage by AG14361 rescued neutrophil-promoted lung cancer progression.Results:An increased number of infiltrating neutrophils was negatively associated with overall survival in lung cancer patients(P<0.001).Neutrophil activation promoted lung cancer cell invasion,migration,and proliferation in vitro,and murine lung cancer growth in vivo.Mechanistically,PARP-1 was shown to be involved in lung cancer cell-induced neutrophil activation to increase MMP-9 expression through interacting and stabilizing ALOX5 by post-translational protein modification(PARylation).Blocking PARP-1 by gene knockdown or AG14361 significantly decreased ALOX5 expression and MMP-9 production,and eliminated neutrophil-mediated lung cancer cell invasion and in vivo tumor growth.Conclusion:We identified a novel mechanism by which PARP-1 mediates lung cancer cell-induced neutrophil activation and PARylates ALOX5 to regulate MMP-9 expression,which exacerbates lung cancer progression.展开更多
Introduction: Collagen is the primary structural protein fibroblasts produce in the skin’s extracellular matrix. Infiltration of neutrophils into the epidermis and dermis by exposure to UV causes collagen damage and ...Introduction: Collagen is the primary structural protein fibroblasts produce in the skin’s extracellular matrix. Infiltration of neutrophils into the epidermis and dermis by exposure to UV causes collagen damage and contributes to photoaging. Methods: To study the combined effect of Lumenato and ceramide in preventing collagen-1 damage induced by phagocytes, we used co-cultures of normal human dermal fibroblasts (fibroblasts) and activated human neutrophils. The present study aimed to determine the protective effect of the combination of Lumenato and ceramide on fibroblast collagen-1 damage induced by neutrophils. Results: Lumenato (in the range of 6.5 - 208 μg/ml) or ceramide (in the range of 0.1 - 50 μM) inhibited the production of superoxides and MPO by TNFα-stimulated neutrophils, as well as the production of NO by LPS-stimulated macrophages in a dose-dependent manner. The combinations of Lumenato and ceramide, in low concentrations, caused synergistic prevention of fibroblasts’ collagen-1 damage induced by TNFα-activated neutrophils, detected by fluorescence immunostaining and WB analysis. MPO activity in the supernatants of the co-cultures was also synergistically inhibited. Adding Lumenato or ceramide singly or in combinations in these low concentrations to the fibroblast cultures did not affect the expression of collagen-1. The combinations of Lumenato or ceramide in these concentrations also caused a synergistic inhibition of NO production by activated macrophages. Conclusions: The results suggest that combining low concentrations of Lumenato and ceramide results in synergistic protection against fibroblasts’ collagen-1 damage induced by neutrophils, thus indicating their possible potential for enhanced skin health.展开更多
Primary and metastatic lung cancers are malignant lung tumors each with of which has a different pathogenesis,although both threaten patient lives.Tumor development and progression involve communication between tumor ...Primary and metastatic lung cancers are malignant lung tumors each with of which has a different pathogenesis,although both threaten patient lives.Tumor development and progression involve communication between tumor cells and the host microenvironment.Neutrophils are the most abundant immune cells in the tumor microenvironment(TME);they participate in the generation of an inflammatory milieu and influence patient survival through their anti-and pro-tumor abilities.Neutrophils can be classified into various categories according to different criteria;frequent categories include N1 antitumor neutrophils and N2 immunosuppressive neutrophils.The antitumor effects of neutrophils are reported to be mediated through a combination of reactive oxygen species,tumor necrosis factor-related apoptosis-inducing ligand,and receptor for advanced glycation end-products–cathepsin G association,as well as the regulation of the activities of other immune cells.There have also been reports that neutrophils can function as tumor promoters that contribute to lung cancer progression and metastasis by influencing processes including carcinogenesis,angiogenesis,cancer cell proliferation,and invasion ability,as well as having similar roles in the lung metastasis of other cancers.The rapid development of nanotechnology has provided new strategies for cancer treatment targeting neutrophils.展开更多
Objective:Liver cancer is a deadly malignancy associated with high mortality and morbidity.Less than 20%of patients with advanced liver cancer respond to a single anti-PD-1 treatment.The high heterogeneity of neutroph...Objective:Liver cancer is a deadly malignancy associated with high mortality and morbidity.Less than 20%of patients with advanced liver cancer respond to a single anti-PD-1 treatment.The high heterogeneity of neutrophils in the tumor immune microenvironment in liver cancer may contribute to resistance to immune checkpoint blockade(ICB).However,the underlying mechanism remains largely unknown.Methods:We established an orthotopic liver cancer model by using transposable elements to integrate the oncogenes Myc and KrasG12Dinto the genome in liver cells from conditional Trp53 null/null mice(pTMK/Trp53^(-/-)).Flow cytometry and immunohistochemistry were used to assess the changes in immune cells in the tumor microenvironment.An ex vivo coculture assay was performed to test the inhibitory effects of tumor-associated neutrophils(TANs)on CD8^(+)T cells.The roles of neutrophils,T cells,and NK cells were validated through antibody-mediated depletion.The efficacy of the combination of neutrophil depletion and ICB was evaluated.Results:Orthotropic pTMK/Trp53^(-/-)mouse liver tumors displayed a moderate response to anti-Ly6G treatment but not PD-1 blockade.Depletion of neutrophils increased the infiltration of CD8^(+)T cells and decreased the number of exhausted T cells in the tumor microenvironment.Furthermore,depletion of either CD8^(+)T or NK cells abrogated the antitumor efficacy of anti-Ly6G treatment.Moreover,the combination of anti-Ly6G with anti-PD-L1 enhanced the infiltration of cytotoxic CD8^(+)T cells and thereafter resulted in a significantly greater decrease in tumor burden.Conclusions:Our data suggest that TANs may contribute to the resistance of liver cancer to ICB,and combining TAN depletion with T cell immunotherapy synergistically increases antitumor efficacy.展开更多
This review is intended to shed new light on the role of neutrophils in colorectal cancer and in the meanwhile emphasiz</span><span style="font-family:Verdana;">e</span><span style="...This review is intended to shed new light on the role of neutrophils in colorectal cancer and in the meanwhile emphasiz</span><span style="font-family:Verdana;">e</span><span style="font-family:""><span style="font-family:Verdana;"> the differences between rectal and colon cancer, strengthen and highlight the possibility of a clinical prognostic and predictive scoring (Sarandria Score). A novel scoring system described in this review can be used as inclusion criteria and as a predictive and prognostic scoring for stage III rectal cancer patients. </span><b><span style="font-family:Verdana;">Background:</span></b><span style="font-family:Verdana;"> Colorectal Cancer (CRC) is a major public health problem, representing the third most commonly diagnosed cancer in males and the second in females. Various studies have reported relevant differences related to CRC primary location site (right-sided colon, left-sided colon, rectum) including response to adjuvant chemotherapy and prognosis. In stage III CRC patients, previous findings showed that higher density of tumor-associated neutrophils (TANs) was associated with better response to 5-FU-based chemotherapy. </span><b><span style="font-family:Verdana;">Main topics:</span></b><span style="font-family:Verdana;"> In this review</span></span><span style="font-family:Verdana;">,</span><span style="font-family:Verdana;"> the current knowledge status on the role of neutrophils in colorectal cancer is assessed, including novel finding discovered by Dr</span><span style="font-family:Verdana;">.</span><span style="font-family:Verdana;"> Nicola Sarandria on the role of neutrophils in rectal cancer. It includes different factors which point to an anti-tumoral role of neutrophils in rectal cancer when in presence of chemotherapeutic agents (such as 5-fluorouracil). The clinical significance of TANs was assessed and whether it can be different depend</span><span style="font-family:Verdana;">ed</span><span style="font-family:""><span style="font-family:Verdana;"> on the location of the primary CRC (right-sided colon, left-sided colon, rectum). </span><b><span style="font-family:Verdana;">Conclusions:</span></b><span style="font-family:Verdana;"> This review officially highlights the possibility of a new clinical prognostic and predictive scoring (Sarandria Score) involving intratumoral neutrophilic infiltration in rectal cancer and the possibility of a new inclusion criteri</span></span><span style="font-family:Verdana;">on</span><span style="font-family:Verdana;"> based on this infiltrate for Stage III rectal cancer patients treated with 5-FU therapy. This review includes knowledge from data published on my medical degree thesis showing that higher levels of TANs densities were associated with better disease-free survival (DFS) in 5-FU treated patients affected by rectal cancer (while it was inversely related in patients without 5-FU therapy). This </span><span style="font-family:Verdana;">i</span><span style="font-family:Verdana;">s also further evidence in support of a possible conceptual division of what is now known as colorectal cancer into two separate entities: colon and rectal cancer.展开更多
Neutrophils play an essential role in the defense against bacterial infections and orchestrate both the innate and adaptive immune responses.With their abundant numbers,diverse function and short life span,these cells...Neutrophils play an essential role in the defense against bacterial infections and orchestrate both the innate and adaptive immune responses.With their abundant numbers,diverse function and short life span,these cells are at the forefront of immune responses,and have gained attention in recent years because of their presence in tumor sites.Neutrophil involvement pertains to tumor cells'ability to construct a suitable tumor microenvironment(TME)that accelerates their own growth and malignancy,by facilitating their interaction with surrounding cells through the circulatory and lymphatic systems,thereby influencing tumor development and progression.Studies have indicated both pro-and anti-tumor properties of infiltrating neutrophils.The TME can exploit neutrophil function,recruitment,and even production,thus resulting in pro-tumor properties of neutrophils,including promotion of genetic instability,tumor cell proliferation,angiogenesis and suppression of anti-tumor or inflammatory response.In contrast,neutrophils can mediate anti-tumor resistance by direct cytotoxicity to the tumor cells or by facilitating anti-tumor functions via crosstalk with T cells.Here,we summarize current knowledge regarding the effects of neutrophil heterogeneity under homeostatic and tumor conditions,including neutrophil phenotype and function,in cancer biology.展开更多
Neutrophils that are mostly related to our body immunity and inflammatory responses are now considered to be an important player in contrast to tumor progression.Along with many controversies,there is much clear evide...Neutrophils that are mostly related to our body immunity and inflammatory responses are now considered to be an important player in contrast to tumor progression.Along with many controversies,there is much clear evidence that shows neutrophils promote the metastasis process via different modes of actions in a different time frame concerning the advancement of cancer.Not only impacting cell proliferation but also they are found to be helping tumor cells in many events like angiogenesis,immune escape,invasion,and finally it leads to cancer metastasis.Tumor microenvironment,key players of cancer growth and progression are also affected by mostly neutrophils and it’s secretory.Tumor associated neutrophils,having certainly different characteristics and expressions,are found high in number around the tumor site and often in pre-metastatic niches.Increased number of premature neutrophils and more neutrophil-extracellular-traps formation is associated with a high degree of metastasis in cancer is observed.As neutrophils has potential role in cancer,more research are warranted in this field to fully understood the overall role of this immune cells and it will helps to open a new way to target neutrophils in therapeutic implications for better cancer treatment.Here we gather currents profile of neutrophils and its role in cancer metastasis and discuss therapeutics strategy.展开更多
Neutrophils are major innate immune effector cells for host defense and have been a topic of active research for their participation in the pathogenesis of autoimmune inflammatory diseases including rheumatoid arthrit...Neutrophils are major innate immune effector cells for host defense and have been a topic of active research for their participation in the pathogenesis of autoimmune inflammatory diseases including rheumatoid arthritis(RA)due to recently discovered neutrophil extracellular trap(NET) formation. NET formation and other mechanisms leading to the release of neutrophil nuclear and cytoplasmic contents are implicated as a source of citrullinated antigens in RA. Further investigations are required to delineate what factors diverge neutrophils from host defense to autoimmune response in RA.展开更多
BACKGROUND:Patients with sepsis often exhibit an acute inflammatory response,followed by an immunosuppressive phase with a poor immune response.However,the underlying mechanisms have not been fully elucidated.METHODS:...BACKGROUND:Patients with sepsis often exhibit an acute inflammatory response,followed by an immunosuppressive phase with a poor immune response.However,the underlying mechanisms have not been fully elucidated.METHODS:We sought to comprehensively characterize the transcriptional changes in neutrophils of patients with sepsis by transcriptome sequencing.Additionally,we conducted a series of experiments,including real-time quantitative polymerase chain reaction(RT-qPCR)and flow cytometry to investigate the role of arginase-1 signaling in sepsis.RESULTS:Through the analysis of gene expression profiles,we identified that the negative regulation of T cell activation signaling was enriched,and the expression of arginase-1 was high in neutrophils from patients with sepsis.Furthermore,we conducted flow cytometry and found that the function of CD8^(+)T cells in septic patients was impaired.Moreover,neutrophils from septic patients inhibited the percentage of polyfunctional effector CD8^(+)T cells through arginase-1.Additionally,the proportions of granzyme B^(+)IFN^(-)γ^(+)CD8^(+)T and TNF^(-)α^(+)IFN^(-)γ^(+)CD8^(+)T cells increased after inhibition of arginase-1 signaling.CONCLUSION:The impaired effector function of CD8^(+)T cells could be restored by blocking arginase-1 signaling in patients with sepsis.展开更多
<span style="font-family:Verdana;">Neutrophils are the most numerous leukocyte in mammals and normally they are the first phagocyte observed in recently damaged or infected tissues. They play a key rol...<span style="font-family:Verdana;">Neutrophils are the most numerous leukocyte in mammals and normally they are the first phagocyte observed in recently damaged or infected tissues. They play a key role </span><span style="font-family:Verdana;">in</span><span style="font-family:;" "=""><span><span style="font-family:Verdana;"> the innate immune responses to </span><i></i></span><i><i><span style="font-family:Verdana;">Leishmania</span></i><span></span></i><span style="font-family:Verdana;"> and several other microorganisms, nonetheless an exacerbated neutrophils activity can generate a harmful response to the host, therefore its turnover rate is very important to maintain the homeostasis and averts the host tissue damage. Both apoptosis followed by phagocytosis by mononuclear phagocytes (eferocytosis) and reverse transmigration have been considered the main processes for the clearance of neutrophils from injured or infected tissues. However, the interaction with </span><i></i></span><i><i><span style="font-family:Verdana;">Leishmania</span></i><span></span></i><span style="font-family:Verdana;"> and other microbes, as well as molecules produced by arthropod vectors such as sandflies saliva can modify the behavior of neutrophils, causing immediate lysis to prolong their life. In fact, as a result of a long course of coevolution, several microorganisms have developed skills to avoid neutrophil effector mechanisms and take advantage of neutrophil clearance pathways to promote their spread in the host’s body. </span><i><i><span style="font-family:Verdana;">Leishmania</span></i><span></span></i><span style="font-family:Verdana;">, </span><i><i><span style="font-family:Verdana;">Chlamydia pneumoniae</span></i><span></span></i><span style="font-family:Verdana;"> and </span><i><i><span style="font-family:Verdana;">Yersinia pestis</span></i><span></span></i><span style="font-family:Verdana;"> for example use the efferocytic Trojan horse process for their dissemination and immune protection, in a different way vaccinia Ankara virus and </span><i><i><span style="font-family:Verdana;">Toxoplasma gondii</span></i><span></span></i><span style="font-family:Verdana;"> exploit the neutrophil reverse transmigration for the same reason. Here we present an overview of some characteristics of neutrophils and their different destinations after interaction with several microorganisms, with an emphasis on </span><i><i><span style="font-family:Verdana;">Leishmania</span></i><span></span></i><span style="font-family:Verdana;"> species. It was also suggested the probable role of neutrophils reverse transmigration as another possible route for the spreading of </span><i><i><span style="font-family:Verdana;">Leishmania</span></i><span></span></i><span style="font-family:Verdana;"> in the visceral leishmaniasis.</span>展开更多
Objective:Neutrophils play an important role in the pathogenesis of rheumatoid arthritis (RA).In this study,we used the adjuvant-induced arthritis murine model to evaluate the efficacy of celastrol on neutrophil-media...Objective:Neutrophils play an important role in the pathogenesis of rheumatoid arthritis (RA).In this study,we used the adjuvant-induced arthritis murine model to evaluate the efficacy of celastrol on neutrophil-mediated inflammation in RA.Methods:Freund's complete adjuvant-induced arthritis was used as the murine model of RA.Celastrol was intraperitoneally administrated daily after onset of the disease.The joint diameter and inflammatory score were evaluated daily during the treatment period.Myeloperoxidase (MPO) and neutrophil elastase (NE) activities were evaluated by immunohistochemical analyses.Quantitative PCR and enzyme-linked immunoabsorbent assay were used to quantify the expression of cytokines.The expression of apoptosis-related proteins Bcl-2,Bax and caspase-3 were evaluated by western blot.Results:Celastrol suppressed inflammation in joints of arthritic mice and diminished the expression of MPO and NE in the joint tissue.Celastrol significantly inhibited the expression of TNFα and IL-6 induced by LPS in neutrophils in a dose-dependent manner in vitro.Moreover,celastrol induced apoptosis of LPS-stimulated neutrophils by increasing the expression of Bax and cleaved caspase-3 while decreasing Bcl-2 expression.Conclusion:Our findings show that celastrol significantly alleviates murine arthritis by modulating the inflammatory activities of neutrophils.These results indicate that celastrol could serve as an alternative or adjunct modality for the treatment of RA.展开更多
Atherosclerosis or fibrosis and cirrhosis undergo chronic inflammation associated with the adhesion between neutrophils and endothelial cells(ECs)that is mediated by their respective cellular adhesive molecules on sti...Atherosclerosis or fibrosis and cirrhosis undergo chronic inflammation associated with the adhesion between neutrophils and endothelial cells(ECs)that is mediated by their respective cellular adhesive molecules on stiffened blood vessel wall or extracellular matrix(ECM)under shear flow[1-3].However,the mechanical dependence of calcium flux and trail formation in neutrophils remains unclear yet in these processes.First,the effect of substrate stiffness through ECs on neutrophil calcium spike was quantified when the individual neutrophils adhered to EC monolayer pre-placed onto stiffness-varied polyacrylamide(PA)substrate(5 or 34.88 kPa)or glass surface.Our data indicated that E-/P-selectins and ICAM-1s on HUVECs and b2-integrins,PSGL-1s,and CD44s on neutrophils were all involved in mediating neutrophil calcium spike in a stiffness-dependent manner,in which the increase of substrate stiffness enhanced the calcium intensity and spike number.Such stiffness-dependent calcium response is associated with selectin-induced b2-integrin activation through Syk/Src signaling pathway and the F-actin/myosin II function.Moreover,tension-activated calcium ion channels displayed critical roles in initiating stiffness-dependent calcium spike [4].Second,the trail formation of neutrophils to ECs monolayer pre-placed onto the same PA substrate were also tested under shear flow.Live fluorescence imaging showed that neutrophils are able to form long membrane tethers during migration and subsequently leave behind membranous long-lasting trails under shear,which are enriched in LFA-1,Mac-1,and CD44.Moreover,the formation of the trails was inhibited by blocking LFA-1s and Mac-1s,suggesting an important role forβ2-integrins in the trial formation.The recruitment of monocytes was inhibited when pre-blocking ICAM-1s on flowing monocytes,indicating that the neutrophil’s trails employβ2-integrin-ICAM-1 binding to recruit the monocytes.Intriguingly,both the length and the area of the trails increase with increasing substrate stiffness,resulting in the enhanced monocyte recruitment.Inhibition of actin binding protein Arp2/3 impairs the trail formation and dramatically decreases the neutrophil-dependent monocyte recruitment.These data provide an insight into understanding how stiffening of vascular wall could regulate the calcium flux of adhered neutrophils and thus the immune responses in atherosclerosis.They also imply that local mechanical microenvironment is remodeled with the migration of neutrophils,leaving the trails presented to induce and regulate monocyte recruitment.All the results are meaningful in elucidating the occurrence and development of atherosclerosis or fibrosis from the viewpoint of mechanotransduction and also for the potential intervention of cardiovascular disease progress.展开更多
AIM:To assess the amount and pattern of reactive oxygen species(ROS)production in diabetic patientderived neutrophils.METHODS:Blood samples from type 2 diabetes mellitus(DM)patients and volunteers(controls)were subjec...AIM:To assess the amount and pattern of reactive oxygen species(ROS)production in diabetic patientderived neutrophils.METHODS:Blood samples from type 2 diabetes mellitus(DM)patients and volunteers(controls)were subjected to neutrophil isolation and the assessment of neutrophil oxidative burst using chemiluminescence assay.Neutrophils were activated by using phorbol myristate acetate(PMA)and neutrophils without activation were kept as a negative control.The chemilu-minescence readings were obtained by transferring cell suspension into a 1.5 m L Eppendorf tube,with PMA and luminol.Reaction mixtures were gently vortexed and placed inside luminometer for a duration of 5 min.RESULTS:Our results showed that in the resting condition,the secretion of ROS in normal non-diabetic individuals was relatively low compared to diabetic patients.However,the time scale observation revealed that the secreted ROS declined accordingly with time in non-diabetic individuals,yet such a reduction was not detected in diabetic patients where at all the time points,the secretion of ROS was maintained at similar magnitudes.This preliminary study demonstrated that ROS production was significantly higher in patients with DM compared to non-diabetic subjects in both resting and activated conditions.CONCLUSION:The respiratory burst activity of neutrophils could be affected by DM and the elevation of ROS production might be an aggravating factor in diabetic-related complications.展开更多
Modification was made on the reversed-phase high performance liquidchromatography（RP-HPLC）with Yue et al’s method as a base.The modified RP-HPLCwas used to detect leukotriene B<sub>4</sub>（LTB<sub&g...Modification was made on the reversed-phase high performance liquidchromatography（RP-HPLC）with Yue et al’s method as a base.The modified RP-HPLCwas used to detect leukotriene B<sub>4</sub>（LTB<sub>4</sub>）and 5-hydroxyeicosatetraenoic acid（5-HETE）.Itwas found that the modified method has the merits of simpler procedures,shortertesting time and more satisfactory efficacy.展开更多
AIM: To explore the effect of indoleamine 2,3-dioxygenase(IDO) on recruitment and chemotaxis function of neutrophils in Aspergillus fumigatus(A.fumigatus) keratitis.METHODS: C57BL/6 mice models of A.fumigatus keratiti...AIM: To explore the effect of indoleamine 2,3-dioxygenase(IDO) on recruitment and chemotaxis function of neutrophils in Aspergillus fumigatus(A.fumigatus) keratitis.METHODS: C57BL/6 mice models of A.fumigatus keratitis were established by inoculating hyphae of A.fumigatus evenly on the corneas.The clinical scores and inflammatory cytokines expression were measured respectively on the 1^(st), 3^(th), 5^(th) day after infection.The 1-MT(1 mg/m L) was administered by gavage to exert an inhibitory effect on IDO during infection.The mice were divided into control group, 1-MT group, A.fumigatus(A.F.) group, and 1-MT+A.F.groups.The corneas were monitored by slit lamp microscopy, and recorded disease scores in 3 d after infection.Myeloperoxidase(MPO) assay was done to evaluate the neutrophils infiltration.Immunofluorescence staining was used to detect the recruitment of neutrophils in murine corneas.The m RNA of inflammatory cytokines was measured with reverse transcription-polymerase chain reaction(RT-PCR).RESULTS: The corneal inflammation and the clinical score reached the peak on the 3;day after the corneal infection.The m RNA of inflammatory cytokines of the A.F.group reached the highest on the 3;day after the infection accordingly.Meanwhile, the results of slit light photography indicated that inhibitors of IDO made inflammation more serious contrasted with the A.F.group on the 3;day.Besides, imunofluorescence staining and MPO indicated that 1-MT enhanced the recruitment, infiltration and chemotaxis of neutrophils obviously in contrast to the A.F.group.RT-PCR indicated that 1-MT increased the expression of CXCL-1, ICAM-1, IL-1β, and IL-8 significantly.CONCLUSION: IDO participates in the pathogenesis of A.fumigatus keratitis and plays an important role in inducing immune protection by inhibiting neutrophils-related inflammatory reaction and suppressing recruitment and chemotaxis of the neutrophils.展开更多
Background: There is a necessity for an establishment of specific markers of oxidative stress for screening in populations of high risk and an estimation of efficiency antioxidative therapies. Objective: Studying the ...Background: There is a necessity for an establishment of specific markers of oxidative stress for screening in populations of high risk and an estimation of efficiency antioxidative therapies. Objective: Studying the activity of antioxidative enzymes at neutrophils in patients with acute coronary syndrome depending on outcome during the year. Methods: Intracellular metabolism of neutrophils was studied in 108 patients, of whom, in 58 persons has been diagnosed acute coronary syndrome, and 50 individuals were without coronary heart disease. Results: In patients with acute coronary syndrome, in comparison with patients without coronary heart disease, growth of production of superoxide anion on background reduction of glutathione reductase activity in neutrophils was revealed. The greatest reduction in parameters of glutathione reductase and catalase at simultaneous growth of activity of NAD(P)H oxidase and myeloperoxidase at neutrophils was observed in patients with fatal outcome during the year. Conclusion: The decrease of efficiency antioxidative protection of neutrophils associates with risk of fatal outcome in patients with acute coronary syndrome.展开更多
Reference genes are essential for studying mRNA expression with quantitative PCR (qPCR). We investigated 11 candidate whole-blood neutrophil reference genes (ACTB, B2M, G6PD, GAPDH, GYPC, HPRT, PGK1, RPL19, SDHA, TFRC...Reference genes are essential for studying mRNA expression with quantitative PCR (qPCR). We investigated 11 candidate whole-blood neutrophil reference genes (ACTB, B2M, G6PD, GAPDH, GYPC, HPRT, PGK1, RPL19, SDHA, TFRC, and YWHAZ) for beef calves, both males and females, with or without selenium supplementation. Initial screening was based on gene expression level (<28 Cq cycles), variability (SD < 1.5 Cq cycles), excluded GYPC and TFRC from further analysis. Expression stability of the remaining genes was evaluated using four software programs: geNorm, NormFinder, BestKeeper, and the comparative delta Cq method. The neutrophil reference genes, YWHAZ, PGK1, and RPL19, consistently ranked among the top four most stable genes under these experimental conditions. The commonly used reference genes, ACTB and HPRT, were not reliable, underscoring the need to validate neutrophil reference genes under different experimental conditions. Multiple reference genes rather than a single gene may provide more robust and reliable results. The best pair of reference genes in whole-blood neutrophils from beef calves overall was PGK1|YWHAZ.展开更多
Neutrophils are granulocytic cytotoxic leukocytes of the innate immune system that activate during acute inflammation. Neutrophils can also persist beyond the acute phase of inflammation to impact the adaptive immune ...Neutrophils are granulocytic cytotoxic leukocytes of the innate immune system that activate during acute inflammation. Neutrophils can also persist beyond the acute phase of inflammation to impact the adaptive immune response during chronic inflammation. In the context of the autoimmune disease, neutrophils modulating T and B cell functions by producing cytokines and chemokines, forming neutrophil extracellular traps, and acting as or priming antigen presentation cells. Thus, neutrophils are actively involved in chronic inflammation and tissue damage in autoimmune disease. Using rheumatoid arthritis as an example, this review focuses on functions of neutrophils in adaptive immunity and the therapeutic potential of these cells in the treatment of autoimmune disease and chronic inflammation.展开更多
基金supported by grants from the National Key R&D Program of China(Grant No.2018YFA0900900)the National Natural Science Foundation of China(Grant Nos.82273334,82203172,81871869,and 81400055)+3 种基金the Jiangsu Province Social Development Key Projects(Grant Nos.BE2020641 and BE2020640)the Xuzhou Medical University Excellent Talent Research Start-up Fund(Grant No.RC20552157)the Jiangsu Province Capability Improvement Project through Science,Technology and Education(Grant No.CXZX202234)funded by the China Postdoctoral Science Foundation(Grant No.2023M732970)。
文摘Objective:Neutrophils are one of the most predominant infiltrating leukocytes in lung cancer tissues and are associated with lung cancer progression.How neutrophils promote lung cancer progression,however,has not been established.Methods:Kaplan–Meier plotter online analysis and tissue immunohistochemistry were used to determine the relationship between neutrophils and overall survival in lung cancer patients.The effect of neutrophils on lung cancer was determined using the Transwell migration assay,a proliferation assay,and a murine tumor model.Gene knockdown was used to determine poly ADPribose polymerase(PARP)-1 function in lung cancer-educated neutrophils.Western blot analysis and gelatin zymography were used to demonstrate the correlation between PARP-1 and matrix metallopeptidase 9(MMP-9).Immunoprecipitation coupled to mass spectrometry(IP/MS)was used to identify the proteins interacting with PARP-1.Co-immunoprecipitation(Co-IP)was used to confirm that PARP-1 interacts with arachidonate 5-lipooxygenase(ALOX5).Neutrophil PARP-1 blockage by AG14361 rescued neutrophil-promoted lung cancer progression.Results:An increased number of infiltrating neutrophils was negatively associated with overall survival in lung cancer patients(P<0.001).Neutrophil activation promoted lung cancer cell invasion,migration,and proliferation in vitro,and murine lung cancer growth in vivo.Mechanistically,PARP-1 was shown to be involved in lung cancer cell-induced neutrophil activation to increase MMP-9 expression through interacting and stabilizing ALOX5 by post-translational protein modification(PARylation).Blocking PARP-1 by gene knockdown or AG14361 significantly decreased ALOX5 expression and MMP-9 production,and eliminated neutrophil-mediated lung cancer cell invasion and in vivo tumor growth.Conclusion:We identified a novel mechanism by which PARP-1 mediates lung cancer cell-induced neutrophil activation and PARylates ALOX5 to regulate MMP-9 expression,which exacerbates lung cancer progression.
文摘Introduction: Collagen is the primary structural protein fibroblasts produce in the skin’s extracellular matrix. Infiltration of neutrophils into the epidermis and dermis by exposure to UV causes collagen damage and contributes to photoaging. Methods: To study the combined effect of Lumenato and ceramide in preventing collagen-1 damage induced by phagocytes, we used co-cultures of normal human dermal fibroblasts (fibroblasts) and activated human neutrophils. The present study aimed to determine the protective effect of the combination of Lumenato and ceramide on fibroblast collagen-1 damage induced by neutrophils. Results: Lumenato (in the range of 6.5 - 208 μg/ml) or ceramide (in the range of 0.1 - 50 μM) inhibited the production of superoxides and MPO by TNFα-stimulated neutrophils, as well as the production of NO by LPS-stimulated macrophages in a dose-dependent manner. The combinations of Lumenato and ceramide, in low concentrations, caused synergistic prevention of fibroblasts’ collagen-1 damage induced by TNFα-activated neutrophils, detected by fluorescence immunostaining and WB analysis. MPO activity in the supernatants of the co-cultures was also synergistically inhibited. Adding Lumenato or ceramide singly or in combinations in these low concentrations to the fibroblast cultures did not affect the expression of collagen-1. The combinations of Lumenato or ceramide in these concentrations also caused a synergistic inhibition of NO production by activated macrophages. Conclusions: The results suggest that combining low concentrations of Lumenato and ceramide results in synergistic protection against fibroblasts’ collagen-1 damage induced by neutrophils, thus indicating their possible potential for enhanced skin health.
基金financially supported by the National Natural Science Foundation of China(31971318,21876205,22027810,and 32101091)China Postdoctoral Science Foundation(2021M690043)+2 种基金the Key-Area Research and Development Program of Guangdong Province(2020B0101020001)the Chinese Academy of Sciences(CAS)Key Research Program for Frontier Sciences(QYZDJSSW-SLH022)the CAS Interdisciplinary Innovation Team,and Big Data Program of PLA General Hospital(2017MBD-016)。
文摘Primary and metastatic lung cancers are malignant lung tumors each with of which has a different pathogenesis,although both threaten patient lives.Tumor development and progression involve communication between tumor cells and the host microenvironment.Neutrophils are the most abundant immune cells in the tumor microenvironment(TME);they participate in the generation of an inflammatory milieu and influence patient survival through their anti-and pro-tumor abilities.Neutrophils can be classified into various categories according to different criteria;frequent categories include N1 antitumor neutrophils and N2 immunosuppressive neutrophils.The antitumor effects of neutrophils are reported to be mediated through a combination of reactive oxygen species,tumor necrosis factor-related apoptosis-inducing ligand,and receptor for advanced glycation end-products–cathepsin G association,as well as the regulation of the activities of other immune cells.There have also been reports that neutrophils can function as tumor promoters that contribute to lung cancer progression and metastasis by influencing processes including carcinogenesis,angiogenesis,cancer cell proliferation,and invasion ability,as well as having similar roles in the lung metastasis of other cancers.The rapid development of nanotechnology has provided new strategies for cancer treatment targeting neutrophils.
基金jointly supported by the National Natural Science Foundation of China(Grant Nos.81972735,82030079,and 81972656)Beijing Natural Science Foundation(Grant No.7212108)+2 种基金Michigan Medicine and PKU-HSC JI(Grant No.BMU2020JI005)Baidu Foundation(Grant No.2020BD015)Ying Shi Foundation。
文摘Objective:Liver cancer is a deadly malignancy associated with high mortality and morbidity.Less than 20%of patients with advanced liver cancer respond to a single anti-PD-1 treatment.The high heterogeneity of neutrophils in the tumor immune microenvironment in liver cancer may contribute to resistance to immune checkpoint blockade(ICB).However,the underlying mechanism remains largely unknown.Methods:We established an orthotopic liver cancer model by using transposable elements to integrate the oncogenes Myc and KrasG12Dinto the genome in liver cells from conditional Trp53 null/null mice(pTMK/Trp53^(-/-)).Flow cytometry and immunohistochemistry were used to assess the changes in immune cells in the tumor microenvironment.An ex vivo coculture assay was performed to test the inhibitory effects of tumor-associated neutrophils(TANs)on CD8^(+)T cells.The roles of neutrophils,T cells,and NK cells were validated through antibody-mediated depletion.The efficacy of the combination of neutrophil depletion and ICB was evaluated.Results:Orthotropic pTMK/Trp53^(-/-)mouse liver tumors displayed a moderate response to anti-Ly6G treatment but not PD-1 blockade.Depletion of neutrophils increased the infiltration of CD8^(+)T cells and decreased the number of exhausted T cells in the tumor microenvironment.Furthermore,depletion of either CD8^(+)T or NK cells abrogated the antitumor efficacy of anti-Ly6G treatment.Moreover,the combination of anti-Ly6G with anti-PD-L1 enhanced the infiltration of cytotoxic CD8^(+)T cells and thereafter resulted in a significantly greater decrease in tumor burden.Conclusions:Our data suggest that TANs may contribute to the resistance of liver cancer to ICB,and combining TAN depletion with T cell immunotherapy synergistically increases antitumor efficacy.
文摘This review is intended to shed new light on the role of neutrophils in colorectal cancer and in the meanwhile emphasiz</span><span style="font-family:Verdana;">e</span><span style="font-family:""><span style="font-family:Verdana;"> the differences between rectal and colon cancer, strengthen and highlight the possibility of a clinical prognostic and predictive scoring (Sarandria Score). A novel scoring system described in this review can be used as inclusion criteria and as a predictive and prognostic scoring for stage III rectal cancer patients. </span><b><span style="font-family:Verdana;">Background:</span></b><span style="font-family:Verdana;"> Colorectal Cancer (CRC) is a major public health problem, representing the third most commonly diagnosed cancer in males and the second in females. Various studies have reported relevant differences related to CRC primary location site (right-sided colon, left-sided colon, rectum) including response to adjuvant chemotherapy and prognosis. In stage III CRC patients, previous findings showed that higher density of tumor-associated neutrophils (TANs) was associated with better response to 5-FU-based chemotherapy. </span><b><span style="font-family:Verdana;">Main topics:</span></b><span style="font-family:Verdana;"> In this review</span></span><span style="font-family:Verdana;">,</span><span style="font-family:Verdana;"> the current knowledge status on the role of neutrophils in colorectal cancer is assessed, including novel finding discovered by Dr</span><span style="font-family:Verdana;">.</span><span style="font-family:Verdana;"> Nicola Sarandria on the role of neutrophils in rectal cancer. It includes different factors which point to an anti-tumoral role of neutrophils in rectal cancer when in presence of chemotherapeutic agents (such as 5-fluorouracil). The clinical significance of TANs was assessed and whether it can be different depend</span><span style="font-family:Verdana;">ed</span><span style="font-family:""><span style="font-family:Verdana;"> on the location of the primary CRC (right-sided colon, left-sided colon, rectum). </span><b><span style="font-family:Verdana;">Conclusions:</span></b><span style="font-family:Verdana;"> This review officially highlights the possibility of a new clinical prognostic and predictive scoring (Sarandria Score) involving intratumoral neutrophilic infiltration in rectal cancer and the possibility of a new inclusion criteri</span></span><span style="font-family:Verdana;">on</span><span style="font-family:Verdana;"> based on this infiltrate for Stage III rectal cancer patients treated with 5-FU therapy. This review includes knowledge from data published on my medical degree thesis showing that higher levels of TANs densities were associated with better disease-free survival (DFS) in 5-FU treated patients affected by rectal cancer (while it was inversely related in patients without 5-FU therapy). This </span><span style="font-family:Verdana;">i</span><span style="font-family:Verdana;">s also further evidence in support of a possible conceptual division of what is now known as colorectal cancer into two separate entities: colon and rectal cancer.
基金A.Y.H. was supported by NRSA Institutional Postdoctoral Training grant T32 (Grant No. 5T32HL066987-20)C.S. was supported by grants from the National Natural Science Foundation of China (Grant No. 82001661)+1 种基金F.X.M. and C.S. were supported by the Haihe Laboratory of Cell Ecosystem Innovation Fund (Grant No. HH22KYZX0019)F.X.M. was supported by the National Natural Science Foundation of China (Grant No. 82171756)
文摘Neutrophils play an essential role in the defense against bacterial infections and orchestrate both the innate and adaptive immune responses.With their abundant numbers,diverse function and short life span,these cells are at the forefront of immune responses,and have gained attention in recent years because of their presence in tumor sites.Neutrophil involvement pertains to tumor cells'ability to construct a suitable tumor microenvironment(TME)that accelerates their own growth and malignancy,by facilitating their interaction with surrounding cells through the circulatory and lymphatic systems,thereby influencing tumor development and progression.Studies have indicated both pro-and anti-tumor properties of infiltrating neutrophils.The TME can exploit neutrophil function,recruitment,and even production,thus resulting in pro-tumor properties of neutrophils,including promotion of genetic instability,tumor cell proliferation,angiogenesis and suppression of anti-tumor or inflammatory response.In contrast,neutrophils can mediate anti-tumor resistance by direct cytotoxicity to the tumor cells or by facilitating anti-tumor functions via crosstalk with T cells.Here,we summarize current knowledge regarding the effects of neutrophil heterogeneity under homeostatic and tumor conditions,including neutrophil phenotype and function,in cancer biology.
文摘Neutrophils that are mostly related to our body immunity and inflammatory responses are now considered to be an important player in contrast to tumor progression.Along with many controversies,there is much clear evidence that shows neutrophils promote the metastasis process via different modes of actions in a different time frame concerning the advancement of cancer.Not only impacting cell proliferation but also they are found to be helping tumor cells in many events like angiogenesis,immune escape,invasion,and finally it leads to cancer metastasis.Tumor microenvironment,key players of cancer growth and progression are also affected by mostly neutrophils and it’s secretory.Tumor associated neutrophils,having certainly different characteristics and expressions,are found high in number around the tumor site and often in pre-metastatic niches.Increased number of premature neutrophils and more neutrophil-extracellular-traps formation is associated with a high degree of metastasis in cancer is observed.As neutrophils has potential role in cancer,more research are warranted in this field to fully understood the overall role of this immune cells and it will helps to open a new way to target neutrophils in therapeutic implications for better cancer treatment.Here we gather currents profile of neutrophils and its role in cancer metastasis and discuss therapeutics strategy.
基金supported by Rheumatology Research Foundation Innovative and Pilot grants and VA Merit Review grant(BX002858).
文摘Neutrophils are major innate immune effector cells for host defense and have been a topic of active research for their participation in the pathogenesis of autoimmune inflammatory diseases including rheumatoid arthritis(RA)due to recently discovered neutrophil extracellular trap(NET) formation. NET formation and other mechanisms leading to the release of neutrophil nuclear and cytoplasmic contents are implicated as a source of citrullinated antigens in RA. Further investigations are required to delineate what factors diverge neutrophils from host defense to autoimmune response in RA.
基金This work was supported by the Research Fund for the Key Laboratory of Anhui Province(KLICD-2022-Z2)the Scientific Research Fund of Anhui Medical University(2011×kj083)the Scientific Research Fund of the First People's Hospital of Hefei(201642).
文摘BACKGROUND:Patients with sepsis often exhibit an acute inflammatory response,followed by an immunosuppressive phase with a poor immune response.However,the underlying mechanisms have not been fully elucidated.METHODS:We sought to comprehensively characterize the transcriptional changes in neutrophils of patients with sepsis by transcriptome sequencing.Additionally,we conducted a series of experiments,including real-time quantitative polymerase chain reaction(RT-qPCR)and flow cytometry to investigate the role of arginase-1 signaling in sepsis.RESULTS:Through the analysis of gene expression profiles,we identified that the negative regulation of T cell activation signaling was enriched,and the expression of arginase-1 was high in neutrophils from patients with sepsis.Furthermore,we conducted flow cytometry and found that the function of CD8^(+)T cells in septic patients was impaired.Moreover,neutrophils from septic patients inhibited the percentage of polyfunctional effector CD8^(+)T cells through arginase-1.Additionally,the proportions of granzyme B^(+)IFN^(-)γ^(+)CD8^(+)T and TNF^(-)α^(+)IFN^(-)γ^(+)CD8^(+)T cells increased after inhibition of arginase-1 signaling.CONCLUSION:The impaired effector function of CD8^(+)T cells could be restored by blocking arginase-1 signaling in patients with sepsis.
文摘<span style="font-family:Verdana;">Neutrophils are the most numerous leukocyte in mammals and normally they are the first phagocyte observed in recently damaged or infected tissues. They play a key role </span><span style="font-family:Verdana;">in</span><span style="font-family:;" "=""><span><span style="font-family:Verdana;"> the innate immune responses to </span><i></i></span><i><i><span style="font-family:Verdana;">Leishmania</span></i><span></span></i><span style="font-family:Verdana;"> and several other microorganisms, nonetheless an exacerbated neutrophils activity can generate a harmful response to the host, therefore its turnover rate is very important to maintain the homeostasis and averts the host tissue damage. Both apoptosis followed by phagocytosis by mononuclear phagocytes (eferocytosis) and reverse transmigration have been considered the main processes for the clearance of neutrophils from injured or infected tissues. However, the interaction with </span><i></i></span><i><i><span style="font-family:Verdana;">Leishmania</span></i><span></span></i><span style="font-family:Verdana;"> and other microbes, as well as molecules produced by arthropod vectors such as sandflies saliva can modify the behavior of neutrophils, causing immediate lysis to prolong their life. In fact, as a result of a long course of coevolution, several microorganisms have developed skills to avoid neutrophil effector mechanisms and take advantage of neutrophil clearance pathways to promote their spread in the host’s body. </span><i><i><span style="font-family:Verdana;">Leishmania</span></i><span></span></i><span style="font-family:Verdana;">, </span><i><i><span style="font-family:Verdana;">Chlamydia pneumoniae</span></i><span></span></i><span style="font-family:Verdana;"> and </span><i><i><span style="font-family:Verdana;">Yersinia pestis</span></i><span></span></i><span style="font-family:Verdana;"> for example use the efferocytic Trojan horse process for their dissemination and immune protection, in a different way vaccinia Ankara virus and </span><i><i><span style="font-family:Verdana;">Toxoplasma gondii</span></i><span></span></i><span style="font-family:Verdana;"> exploit the neutrophil reverse transmigration for the same reason. Here we present an overview of some characteristics of neutrophils and their different destinations after interaction with several microorganisms, with an emphasis on </span><i><i><span style="font-family:Verdana;">Leishmania</span></i><span></span></i><span style="font-family:Verdana;"> species. It was also suggested the probable role of neutrophils reverse transmigration as another possible route for the spreading of </span><i><i><span style="font-family:Verdana;">Leishmania</span></i><span></span></i><span style="font-family:Verdana;"> in the visceral leishmaniasis.</span>
基金the National Natural Science Foundation of China(Grant number 81430099)International S&T Cooperation Program of China(Grant number 2014DFA32950)Beijing Municipal Commission of Education(Grant number 521/0101312).
文摘Objective:Neutrophils play an important role in the pathogenesis of rheumatoid arthritis (RA).In this study,we used the adjuvant-induced arthritis murine model to evaluate the efficacy of celastrol on neutrophil-mediated inflammation in RA.Methods:Freund's complete adjuvant-induced arthritis was used as the murine model of RA.Celastrol was intraperitoneally administrated daily after onset of the disease.The joint diameter and inflammatory score were evaluated daily during the treatment period.Myeloperoxidase (MPO) and neutrophil elastase (NE) activities were evaluated by immunohistochemical analyses.Quantitative PCR and enzyme-linked immunoabsorbent assay were used to quantify the expression of cytokines.The expression of apoptosis-related proteins Bcl-2,Bax and caspase-3 were evaluated by western blot.Results:Celastrol suppressed inflammation in joints of arthritic mice and diminished the expression of MPO and NE in the joint tissue.Celastrol significantly inhibited the expression of TNFα and IL-6 induced by LPS in neutrophils in a dose-dependent manner in vitro.Moreover,celastrol induced apoptosis of LPS-stimulated neutrophils by increasing the expression of Bax and cleaved caspase-3 while decreasing Bcl-2 expression.Conclusion:Our findings show that celastrol significantly alleviates murine arthritis by modulating the inflammatory activities of neutrophils.These results indicate that celastrol could serve as an alternative or adjunct modality for the treatment of RA.
基金supported by National Natural Science Foundation of China Grant( 31627804,91642203, 11772345,91539119)Chinese Academy of Sciences Strategic Priority Research Program ( XDB22040101)Frontier Science Key Project( QYZDJ-SSWJSC018)
文摘Atherosclerosis or fibrosis and cirrhosis undergo chronic inflammation associated with the adhesion between neutrophils and endothelial cells(ECs)that is mediated by their respective cellular adhesive molecules on stiffened blood vessel wall or extracellular matrix(ECM)under shear flow[1-3].However,the mechanical dependence of calcium flux and trail formation in neutrophils remains unclear yet in these processes.First,the effect of substrate stiffness through ECs on neutrophil calcium spike was quantified when the individual neutrophils adhered to EC monolayer pre-placed onto stiffness-varied polyacrylamide(PA)substrate(5 or 34.88 kPa)or glass surface.Our data indicated that E-/P-selectins and ICAM-1s on HUVECs and b2-integrins,PSGL-1s,and CD44s on neutrophils were all involved in mediating neutrophil calcium spike in a stiffness-dependent manner,in which the increase of substrate stiffness enhanced the calcium intensity and spike number.Such stiffness-dependent calcium response is associated with selectin-induced b2-integrin activation through Syk/Src signaling pathway and the F-actin/myosin II function.Moreover,tension-activated calcium ion channels displayed critical roles in initiating stiffness-dependent calcium spike [4].Second,the trail formation of neutrophils to ECs monolayer pre-placed onto the same PA substrate were also tested under shear flow.Live fluorescence imaging showed that neutrophils are able to form long membrane tethers during migration and subsequently leave behind membranous long-lasting trails under shear,which are enriched in LFA-1,Mac-1,and CD44.Moreover,the formation of the trails was inhibited by blocking LFA-1s and Mac-1s,suggesting an important role forβ2-integrins in the trial formation.The recruitment of monocytes was inhibited when pre-blocking ICAM-1s on flowing monocytes,indicating that the neutrophil’s trails employβ2-integrin-ICAM-1 binding to recruit the monocytes.Intriguingly,both the length and the area of the trails increase with increasing substrate stiffness,resulting in the enhanced monocyte recruitment.Inhibition of actin binding protein Arp2/3 impairs the trail formation and dramatically decreases the neutrophil-dependent monocyte recruitment.These data provide an insight into understanding how stiffening of vascular wall could regulate the calcium flux of adhered neutrophils and thus the immune responses in atherosclerosis.They also imply that local mechanical microenvironment is remodeled with the migration of neutrophils,leaving the trails presented to induce and regulate monocyte recruitment.All the results are meaningful in elucidating the occurrence and development of atherosclerosis or fibrosis from the viewpoint of mechanotransduction and also for the potential intervention of cardiovascular disease progress.
文摘AIM:To assess the amount and pattern of reactive oxygen species(ROS)production in diabetic patientderived neutrophils.METHODS:Blood samples from type 2 diabetes mellitus(DM)patients and volunteers(controls)were subjected to neutrophil isolation and the assessment of neutrophil oxidative burst using chemiluminescence assay.Neutrophils were activated by using phorbol myristate acetate(PMA)and neutrophils without activation were kept as a negative control.The chemilu-minescence readings were obtained by transferring cell suspension into a 1.5 m L Eppendorf tube,with PMA and luminol.Reaction mixtures were gently vortexed and placed inside luminometer for a duration of 5 min.RESULTS:Our results showed that in the resting condition,the secretion of ROS in normal non-diabetic individuals was relatively low compared to diabetic patients.However,the time scale observation revealed that the secreted ROS declined accordingly with time in non-diabetic individuals,yet such a reduction was not detected in diabetic patients where at all the time points,the secretion of ROS was maintained at similar magnitudes.This preliminary study demonstrated that ROS production was significantly higher in patients with DM compared to non-diabetic subjects in both resting and activated conditions.CONCLUSION:The respiratory burst activity of neutrophils could be affected by DM and the elevation of ROS production might be an aggravating factor in diabetic-related complications.
文摘Modification was made on the reversed-phase high performance liquidchromatography（RP-HPLC）with Yue et al’s method as a base.The modified RP-HPLCwas used to detect leukotriene B<sub>4</sub>（LTB<sub>4</sub>）and 5-hydroxyeicosatetraenoic acid（5-HETE）.Itwas found that the modified method has the merits of simpler procedures,shortertesting time and more satisfactory efficacy.
基金Supported by the National Natural Science Foundation of China (No.81870632)the Youth National Natural Science Foundation of China (No.81700800)the Natural Science Foundation of Shandong Province (No.ZR2017MH008)。
文摘AIM: To explore the effect of indoleamine 2,3-dioxygenase(IDO) on recruitment and chemotaxis function of neutrophils in Aspergillus fumigatus(A.fumigatus) keratitis.METHODS: C57BL/6 mice models of A.fumigatus keratitis were established by inoculating hyphae of A.fumigatus evenly on the corneas.The clinical scores and inflammatory cytokines expression were measured respectively on the 1^(st), 3^(th), 5^(th) day after infection.The 1-MT(1 mg/m L) was administered by gavage to exert an inhibitory effect on IDO during infection.The mice were divided into control group, 1-MT group, A.fumigatus(A.F.) group, and 1-MT+A.F.groups.The corneas were monitored by slit lamp microscopy, and recorded disease scores in 3 d after infection.Myeloperoxidase(MPO) assay was done to evaluate the neutrophils infiltration.Immunofluorescence staining was used to detect the recruitment of neutrophils in murine corneas.The m RNA of inflammatory cytokines was measured with reverse transcription-polymerase chain reaction(RT-PCR).RESULTS: The corneal inflammation and the clinical score reached the peak on the 3;day after the corneal infection.The m RNA of inflammatory cytokines of the A.F.group reached the highest on the 3;day after the infection accordingly.Meanwhile, the results of slit light photography indicated that inhibitors of IDO made inflammation more serious contrasted with the A.F.group on the 3;day.Besides, imunofluorescence staining and MPO indicated that 1-MT enhanced the recruitment, infiltration and chemotaxis of neutrophils obviously in contrast to the A.F.group.RT-PCR indicated that 1-MT increased the expression of CXCL-1, ICAM-1, IL-1β, and IL-8 significantly.CONCLUSION: IDO participates in the pathogenesis of A.fumigatus keratitis and plays an important role in inducing immune protection by inhibiting neutrophils-related inflammatory reaction and suppressing recruitment and chemotaxis of the neutrophils.
文摘Background: There is a necessity for an establishment of specific markers of oxidative stress for screening in populations of high risk and an estimation of efficiency antioxidative therapies. Objective: Studying the activity of antioxidative enzymes at neutrophils in patients with acute coronary syndrome depending on outcome during the year. Methods: Intracellular metabolism of neutrophils was studied in 108 patients, of whom, in 58 persons has been diagnosed acute coronary syndrome, and 50 individuals were without coronary heart disease. Results: In patients with acute coronary syndrome, in comparison with patients without coronary heart disease, growth of production of superoxide anion on background reduction of glutathione reductase activity in neutrophils was revealed. The greatest reduction in parameters of glutathione reductase and catalase at simultaneous growth of activity of NAD(P)H oxidase and myeloperoxidase at neutrophils was observed in patients with fatal outcome during the year. Conclusion: The decrease of efficiency antioxidative protection of neutrophils associates with risk of fatal outcome in patients with acute coronary syndrome.
文摘Reference genes are essential for studying mRNA expression with quantitative PCR (qPCR). We investigated 11 candidate whole-blood neutrophil reference genes (ACTB, B2M, G6PD, GAPDH, GYPC, HPRT, PGK1, RPL19, SDHA, TFRC, and YWHAZ) for beef calves, both males and females, with or without selenium supplementation. Initial screening was based on gene expression level (<28 Cq cycles), variability (SD < 1.5 Cq cycles), excluded GYPC and TFRC from further analysis. Expression stability of the remaining genes was evaluated using four software programs: geNorm, NormFinder, BestKeeper, and the comparative delta Cq method. The neutrophil reference genes, YWHAZ, PGK1, and RPL19, consistently ranked among the top four most stable genes under these experimental conditions. The commonly used reference genes, ACTB and HPRT, were not reliable, underscoring the need to validate neutrophil reference genes under different experimental conditions. Multiple reference genes rather than a single gene may provide more robust and reliable results. The best pair of reference genes in whole-blood neutrophils from beef calves overall was PGK1|YWHAZ.
基金Supported by The National Institute of Arthritis and Muscu-loskeletal and Skin Diseases of the National Institutes of Health(NIAMS-NIH)No.1R01AR063132
文摘Neutrophils are granulocytic cytotoxic leukocytes of the innate immune system that activate during acute inflammation. Neutrophils can also persist beyond the acute phase of inflammation to impact the adaptive immune response during chronic inflammation. In the context of the autoimmune disease, neutrophils modulating T and B cell functions by producing cytokines and chemokines, forming neutrophil extracellular traps, and acting as or priming antigen presentation cells. Thus, neutrophils are actively involved in chronic inflammation and tissue damage in autoimmune disease. Using rheumatoid arthritis as an example, this review focuses on functions of neutrophils in adaptive immunity and the therapeutic potential of these cells in the treatment of autoimmune disease and chronic inflammation.
