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Caffeic acid and quercetin exert caspases-independent apoptotic effects on Leishmania major promastigotes, and reactivate the death of infected phagocytes derived from BALB/c mice

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Asian Pacific Journal of Tropical Biomedicine 2017年第4期7卷 321-331页

作者： Radia Belkhelfa-Slimani Bahia Djerdjouri Laboratory of Cellular and Molecular Biology Faculty of Biological SciencesUniversity of Sciences and Technology Houari BoumedieneAlgiersAlgeria

Objective: To investigate the leishmanicidal effects of two antioxidants, caffeic acid and quercetin on Leishmania major(L.major) promastigotes in vitro, and their immunomodulatory effects on infected phagocytes derived from susceptible BALB/c mice.Methods: Caffeic acid and quercetin-induced cell death was examined by Pi-Hoechst double staining of L.major promastigotes and MTT assay, in the presence or absence of protease inhibitors in vitro.Caffeic acid or quercetin were administered subcutaneously to BALB/c mice infected with L.major promastigotes through a dorsal air pouch.Nitric oxide and superoxide anion production by phagocytes infiltrating the air pouch and the expression of inducible nitric oxide synthase(i NOS), tumor necrosis factor alpha(TNF-a) and nuclear factor kappa B in the air pouch membrane were therefore evaluated using appropriate methods.Results: Caffeic acid and quercetin displayed a dose-dependent cytotoxic effect against L.major promastigotes, and induced cell death via caspases-independent pathways.In vivo, L.major promastigotes inoculation into air pouch cavity of BALB/c mice leads to a sequential influx of neutrophils(hours), followed by macrophages(days).Results showed that L.major delayed apoptosis of infected neutrophils and macrophages by the cleavage of the nuclear factor kappa B p65^(RelA) subunit, and persisted by inhibiting TNF-a and i NOS expression and reactive oxygen species generation.Caffeic acid or quercetin restored reactive oxygen species production and TNF-a-induced i NOS activity, and abrogate apoptosis delay of infected phagocytes.Conclusions: The leishmanicidal effect of caffeic acid and quercetin on promastigotes and amastigotes, as well as reactivation of infected phagocytes apoptosis, suggested a potential therapeutic role against cutaneous leishmaniasis.

关键词： Apoptosis Leishmania major phagocytes iNOS NF-kB TNF-a

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Functional Activity of Circulating phagocytes as Potential Pretreatment Marker of Tumor Drug Resistance

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Journal of Biosciences and Medicines 2019年第7期7卷 1-15页

作者： Iryna V. Prokhorova Olga V. Yurchenko Olga N. Pyaskovskaya Igor N. Todor Galina I. Solyanik Laboratory of Molecular and Cellular Mechanisms of Metastasis R.E. Kavetsky Institute of Experimental Pathology Oncology and Radiobiology of NASU Kyiv Ukraine Department of Monitoring of Tumor Process and Therapy Design R.E. Kavetsky Institute of Experimental Pathology Oncology and Radiobiology of NASU Kyiv Ukraine

The aim of this work was a study of the functional activity of neutrophils and peripheral blood monocytes in rats with the transplanted Walker carcinosarcoma as potential predictors of the sensitivity of this tumor to doxorubicin treatment. This study provides an evidence that such indices of the functional activity of circulating phagocytes of the tumor-bearing rats as the quantity and the phagocytosis intensity of monocytes, as well as the intensity of ROS production by monocytes and neutrophils, may reflect the degree of sensitivity of the tumor to doxorubicin. So it was shown that the growth of the resistant tumor caused a significant increase of the number of circulating phagocytic cells and the intensity of phagocytosis by more than 100% (p phagocytes, an assessment of the indices of their functional activity before each subsequent course can be considered as a pretreatment assessment. Meanwhile, further studies are needed to determine the spectrum of malignant neoplasms for which the degree of tumor drug resistance correlates with the functional activity of circulating phagocytes.

关键词： Circulating phagocytes Phagocytic Activity ROS Production Tumor Drug Resistance

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The epididymal immune balance:a key to preserving male fertility

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Asian Journal of Andrology 2019年第6期21卷 531-539页

作者： Allison Voisin Fabrice Saez Joel R Drevet Rachel Guiton Team Mechanisms of Posttesticular Infertility GReD LaboratoryCNRS UMR 6293-INSERM U1103 University of Clermont Auvergne Clermont-Ferrand 63001France

Up to 15%of male in fertility has an immuno logical origi n,either due to repetitive infections or to autoimmune resp on ses mainly affecting the epididymis,prostate,and testis.Clinical observations and epidemiological data clearly contradict the idea that the testis confers immune protection to the whole male genital tract.As a consequenee,the epididymis,in which posttesticular spermatozoa mature and are stored,has raised some in terest in recent years when it comes to its immune mecha nisms.In deed,sperm cells are produced at puberty,long after the establishment of self-toleranee,and they possess unique surface proteins that cannot be recognized as self.These are potential targets of the immune system,with the risk of inducing autoantibodies and consequently male infertility.Epididymal immunity is based on a finely tuned equilibrium between efficient immune responses to pathogens and strong tolerance to sperm cells.These processes rely on incompletely described molecules and cell types.This review compiles recent studies focusing on the immune cell types populating the epididymis,and proposes hypothetical models of the organization of epididymal immunity with a special emphasis on the immune response,while also discussing important aspects of the epididymal immune regulation such as tolerance and tumour control.

关键词： epididymis immune response lymphocytes mononuclear phagocytes toleranee

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Leukocyte adhesion defect:Where do we stand circa 2019?

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Genes & Diseases 2020年第1期7卷 107-114页

作者： Jhumki Das Avinash Sharma Ankur Jindal Vaishali Aggarwal Amit Rawat Allergy Immunology Unit Department of PediatricsAdvanced Pediatrics CentrePostgraduate Institute of Medical Education and ResearchChandigarhIndia Department of Pediatrics Dr Rajendra Prasad Government Medical CollegeTandaKangraHimachal PradeshIndia

Migration of polymorphonuclear leukocytes from bloodstream to the site of inflammation is an important event required for surveillance of foreign antigens.This trafficking of leukocytes from bloodstream to the tissue occurs in several distinct steps and involves several adhesion molecules.Defect in adhesion of leukocytes to vascular endothelium affecting their subsequent migration to extravascular space gives rise to a group of rare primary immunodeficiency diseases(PIDs)known as Leukocyte Adhesion Defects(LAD).Till date,four classes of LAD are discovered with LAD I being the most common form.LAD I is caused by loss of function of common chain,cluster of differentiation(CD)18 of β2 integrin family.These patients suffer from life-threatening bacterial infections and in its severe form death usually occurs in childhood without bone marrow transplantation.LAD II results from a general defect in fucose metabolism.These patients suffer from less severe bacterial infections and have growth and mental retardation.Bombay blood group phenotype is also observed in these patients.LAD III is caused by abnormal integrin activation.LAD III patients suffer from severe bacterial and fungal infections.Patients frequently show delayed detachment of umbilical cord,impaired wound healing and increased tendency to bleed.LAD IV is the most recently described class.It is caused by defects in β2 and α4β1 integrins which impairs lymphocyte adhesion.LAD IV patients have monogenic defect in cystic-fibrosis-transmembraneconductance-regulator(CFTR)gene,resulting in cystic fibrosis.Pathophysiology and genetic etiology of all LAD syndromes are discussed in detail in this paper.

关键词： Neutrophilic defect Neutrophilic leukocytosis Phagocyte rolling phagocytes Primary immunodeficiency disorders

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Introduction of antineoplastic drug NSC631570 in an inpatient and outpatient setting:Comparative evaluation of biological effects

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Asian Journal of Pharmaceutical Sciences 2016年第2期11卷 308-317页

作者： Mariia Rudyk Olexander Fedorchuk Yaroslav Susak Yaroslav Nowicky Larysa Skivka Microbiology and General Immunology Department ESC“Institute of Biology”Taras Shevchenko National University of KyivUkraine R.E.Kavetsky Institute of Experimental Pathology Oncology and RadiobiologyNational Academy of Sciences of UkraineUkraine O.O Bogomolets National Medical University KyivUkraine Ukrainian Anticancer Institute ViennaAustria

The aim of this study is to evaluate the effect of moderate physical exercise and treatment time on the organism’s response to NSC631570. The sensitivity of circulating phagocytes to the drug at different times of day was estimated in in vitro experiments. NSC631570 was administered intravenously to healthy volunteers (eleven men, 23 ± 2 years) in a single therapeutic dose in an inpatient and an outpatient setting. Blood samples were obtained before the drug administration, 30 min after the drug injection and every fourth hour throughout the 24 hour period. Biochemical parameters were determined using the hematological analyzer. Flow cytometry was used to evaluate phagocyte metabolism. Treatment of circulating phagocytes with NSC631570 in vitro resulted in an increase in ROS production along with a decrease in their phagocytic activity, most expressed in the morning time. Drug injection to sedentary persons resulted in pro-inflammatory metabolic polarization of circulating phagocytes. Introduction of NSC631570 to active persons was accompanied by a significant increase in phagocyte endocytosis along with a decrease in the daily mean of ROS generation. Significant oscillation (but in the normal ranges) of urea, creatinine, alanine aminotransferase and aspartate aminotransferase after NSC631570 introduction in the outpatient setting was shown during the day. Physical activity interferes with immunomodulatory action of NSC631570 and abrogates pro-inflammatory shift of circulating phagocytes. Biochemical parameters of blood from patients treated with NSC631570 in the outpatient setting must be interpreted cautiously considering the effect of physical activity on some metabolic biomarkers.

关键词： NSC631570 Physical exercise Blood biochemical parameters phagocytes functional polarization

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Reactive astrocytes function as phagocytes and inflammation mediators through Axl/STAT1/ABCA1 pathway after traumatic brain injury

Reactive astrocytes function as phagocytes and inflammation ...

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第十四届中国医师协会神经外科医师年会

作者：周航

Astrocytes and microglia become reactive following traumatic brain injury（TBI）. However, the coordination of this reactivity and its relation to pathophysiology are poorly understood. Here we show that reactive astrocytes function as phagocytes and inflammation mediators after TBI via Axl/STAT1/ABCA1 pathway. Following TBI, phagocytic astrocytes are observed within the peri-injury region forming a neuroprotective astrocyte scar during the later stage（3 d,7 d）, and phagocytic microglia are mainly observed within the injury core during the earlier stage of TBI（1 d）. This change in astrocytes phenotype results from Axl upregulation mediated by microglia-derived gas6. In mice, Axlenhancement and Axl-inhibition lead to reduced and increased neuronal damage respectively by influencing transformation of astrocyte phenotype and mediation of neuroinflammation, and upregulation of Axl is sufficient for enhancement of phagocytosis in vitro. Inhibition of microglia results in fewer phagocytic astrocytes and diminished neuroprotection in later stage of TBI. Together, these findings suggest that astrocytes are transformed into a phagocytic phenotype by active microglia through Axl/STAT1/ABCA1 pathway and contribute to restricting tissue damage and mediating neuroinflammation.

关键词： Reactive astrocytes phagocytes Axl pathway Traumatic brain injury

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Cooperative apoptosis of coelomocytes of the holothurian <i>Eupentacta fraudatrix</i>and its modulation by dexamethasone

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Advances in Bioscience and Biotechnology 2013年第9期4卷 908-917页

作者： Olga A. Zaika Lyudmila S. Dolmatova Il’ichev Pacific Oceanological Institute Far Eastern Branch Russian Academy of Sciences Vladivostok Russia

The capacities of phagocytes of subpopulation P1 (F) and morula cells (MC) of holothurian Eupentacta fraudatrix to modulate apoptosis of each other as well as cytokine-dependent mechanisms and hormonal regulation of these cells’s interaction were studied. The 18-h treatment of F with supernatants, obtained after centrifugation of MC preincubated for 3 h with phosphate buffered saline (PSB) at the temperature of 22℃?(SMC3) resulted in a significant growth of apoptosis level. A 30-min incubation of F with supernatants of MC, preincubated for 24 h (SMC24), on the contrary, reduced the apoptosis level and increased the level of interleukine-1α (IL-1α)-like substances, and 24-h incubation did not influence apoptosis and reduced level of IL-1α-like substances. Thus, proapoptotic effects of MC’s supernatants in F inversely depended on time of their preincubation with PSB and directly on time of incubation with F. Additionally, this effect was opposite to variations in the level of IL-1α-like substances. The level of apoptosis declined after 30 min of incubation but elevated after 24 h at the inverse treatment of MC with supernatant, obtained after preincubation of F during 24 h (SF24). The level of IL-1α-like substances dropped after 30 min and insignificantly decreased after 24 h. Hence, SF24 proapoptotic effect directly depended on time of incubation with MC and did not correspond to variations in the level of IL-1α-like substances. 100 μM dexamethasone stimulated apoptosis in F and MC in an inverse time-dependent manner during 24-h preincubation, and supernatants of cell suspensions obtained after such preincubations, stimulated apoptosis and reduced the IL-1α-like substances level in target cells at both types of interaction. IL-1α-like substances are?supposed to be mediators for MC’s effects in F, but not for F’s action on MC. In holothurians, steroid hormones apparently may participate in the regulation of the immune response and cell cooperation.

关键词： phagocytes Morula Cells Cell Cooperation Dexamethasone Apoptosis Cytokines

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Granulocyte Colony—Stimulating Factor Multiplies Normal Blood ROS Generation at Less than 1 µg/l

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Optics and Photonics Journal 2018年第1期8卷 1-10页

作者： Thomas Stief Institute of Laboratory Medicine and Pathobiochemistry University Hospital of Giessen & Marburg Giessen & Marburg Germany

Background: The neutrophils (PMN) are our main blood cells to combat fungi, bacteria, and fibrin. For normal function, an activated PMN generates a certain concentration of reactive oxygen species (ROS). If the generated blood ROS concentration is too low, then fungi, bacteria or fibrin might threaten the life of the patient, and it could be of great medical interest to stimulate PMN by physiologic drugs. Granulocyte-Colony Stimulating Factor (G-CSF) is a cell hormone that increases the cell number of PMN and that stimulates the individual PMN. The blood ROS generation assay (BRGA) is an innovative physiologic test to monitor the ROS generation of PMN in blood. Here the ROS generating action of G-CSF on normal PMN is quantified. Material and Methods: 40 μl 0 - 10.3 ng/ml (final conc.) G-CSF (in 5% human albumin) in black Brand? 781608 high quality polystyrene F-microwells was incubated in triplicate with 125 μl Hanks’ balanced salt solution (HBSS;modified without phenol red) and 10 μl normal citrated blood. Immediately (BRGA) or after 60 min (BRGA-60-) 10 μl 5 mM luminol sodium salt in 0.9% NaCl and 10 μl 0 or 36 μg/ml zymosan A in 0.9% NaCl was added. The photons were counted within 0 - 318 min (37°C) in a photons-multiplying microtiter plate luminometer. At about 0.5 t-maxn (0.5 fold the time to normal maximum) the approx. SC200 of G-CSF was determined. Results and Discussion: The approx. SC200 of G-CSF on normal blood ROS generation was 0.2 μg/l (=20 IU/ml). In clinical situations where an increased blood ROS generation is pharmacologically required, few micrograms of G-CSF could be a sufficient dosage for an adult patient. The BRGA helps to find out the correct stimulating G-CSF dosage for each individual. An enhanced PMN function could favor a better clinical outcome in situations of wanted increase of the innate immunology or in cellular fibrinolysis. G-CSF plasma concentrations of 0.1 - 1 μg/l might favor singlet oxygen generation without immunosuppression or cell fragment-induced thr

关键词： Singlet Oxygen (1ΔO2) Reactive Oxygen Species (ROS) Excited Carbonyl (R-C = O*) Photon (hν) phagocytes Neutrophils (PMN) BRGA G-CSF

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Glucose initially inhibits and later stimulates blood ROS generation

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Journal of Diabetes Mellitus 2013年第1期3卷 15-21页

作者： Thomas Stief Institute of Laboratory Medicine and Pathobiochemistry Central Laboratory of the University Hospital Marburg Germany

Background: Glucose is the main substrate for the generation of NADPH, the cofactor of the oxidative burst enzyme NADPH-oxidase of blood neutrophils. Changes in blood glucose are thus expected to modify the generation of reactive oxygen species (ROS). The new blood ROS generation assay (BRGA) quantifies ROS changes induced by blood glucose concentrations as they are found in diabetes mellitus. Material and Methods: Citrated or EDTA blood of 6 healthy donors were analyzed in the BRGA: 10 μl sample in black polystyrene F-microwells (Brand 781608) were incubated in triplicate with 125 μl Hanks’ balanced salt solution, 40 μl 0 - 200 mM glucose in 0.9% NaCl (final added conc.: 0 - 41 mM;final basal glucose conc.: about4 mM), 10 μl5 mMluminol, and 10 μl zymosan A (final conc.: 1.9 μg/ml) in 0.9% NaCl. The plates were measured within 0 - 250 min (37℃) in a photons-multiplyer microtiter plate luminometer (LUmo) with an integration time of 1 s. Results: Up to about 30 min reaction time the mean ROS generation was 50% inhibited by about1 mMadded glucose (= approx. IC50). At ≥80 min reaction time (possibly necessary for full phosphorylation of glucose to glucose-6-phosphate (G6P), the substrate metabolized by G6P-dehydrogenase to generate NADPH, the cofactor of the NADPH-oxidase) the mean ROS generation approximately doubled at about1 mMadded glucose (= approx. SC200) in citrated blood. Discussion: Elevated glucose concentrations not only increase systemic thrombin generation, they can also diminish cellular fibrinolysis and increase systemic inflammation, resulting in a chronic pro-thrombotic state. The fascinating importance of NADPH-oxidases not only in phagocytes but also in the beta cells of pancreas points towards a new pathogenesis explication of diabetes mellitus type 1: whatever stimulus (e.g. a pancreas-tropic virus) could activate the beta cell’s autodestructive NADPH-oxidase.

关键词： Reactive Oxygen Species ROS Neutrophils phagocytes Blood ROS Generation Assay BRGA NADPH-Oxidase Glucose

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解淀粉芽孢杆菌对小鼠肠道健康和免疫功能的影响

解淀粉芽孢杆菌对小鼠肠道健康和免疫功能的影响

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中国畜牧兽医学会动物微生态学分会第十一次全国学术研讨会...

作者：杜威黄琴付爱坤毛予龙李雅丽余东游李卫芬浙江大学动物科学学院饲料研究所动物营养与饲料科学重点实验室

The present study investigated the effects of dietary supplementation of Bacillus amyloliquefaciens SC06 on intestinal environment as well as intestinal,humoral and cellular response in BALB/c mice.The results showed that dietary Bacillus amyloliquefaciens SC06 could improve intestinal health through affecting bacterial enzyme activities in the caecal and small intestinal digesta.In addition,this Bacillus strain could significantly increase the level of cytokines,β-defensins and the activities of myeloperoxidase(MPO)and secretory phospholipase A2(sPLA2)(P0.05).Oral administration of Bacillus amyloliquefaciens SC06 could significantly increase serum antibody titers and the number of plaque-forming cells.(PFC)(pphagocytes,the phagocytic function of the mononuclear phagocyte system(MPS),NK cell-mediated cytotoxicity,the percentage of IFN-γ-producing CD4 and CD8 T cells(P<0.01),the percentages of CD3 and CD4cell(P<0.05).However,it had no effect on lymphocyte proliferation induced by T cell mitogen and B cell mitogen.All above,our results indicated that dietary Bacillus amyloliquefaciens SC06 could improve intestinal health through improving microbita composition,and showed positive effect on intestinal,the humoral and the cellular immune function by activating the Th1-type immune response.

关键词： Bacillus amyloliquefaciens bacterial enzyme activities intestinal immune cellular immune phagocytes spleen lymphocyte subsets

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