scholarly journals The role of eosinophils and corresponding cytokines and chemokines asthma-COPD overlap (ACO) patients

2020 ◽  
Author(s):  
Qinglan Li ◽  
Liang Lu ◽  
Shiyang Geng ◽  
Huiyun Zhang ◽  
Xin Li ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Healthy Volunteers ◽  
Clinical Response ◽  
Acute Exacerbation ◽  
Plasma Levels ◽  
Flow Cytometry Analysis ◽  
Cytokines And Chemokines ◽  
Acute Exacerbations ◽  
Treatment Responses

Abstract Background: ACO has been characterized as a kind of clinical disease with overlap symptoms of asthma and COPD. However, little is known of the role of eosinophils and corresponding cytokines and chemokines in ACO patients with different treatment responses. Methods: To evaluate factors which associates with different treatment responses in patients with ACO. In the present study, we investigated the eosinophils proportion of peripheral blood from ACO patients with acute exacerbation (AE) after treatment, ACO patients with clinical response (CR) after treatment, and healthy volunteers (HV) by using flow cytometry analysis. The plasma levels of corresponding cytokines and chemokines from the three groups were evaluated by ELISA.Results: The results showed that ACO patients that had acute exacerbation have relatively lower eosinophils proportions compared to healthy volunteers but have higher eosinophils inflammation compared with patients with clinical response. The percentage of NK1R+ expression population eosinophils was also decreased. Further analysis revealed ACO patients that had acute exacerbation also have relatively higher plasma levels of cytokines and chemokines compared to patients with clinical response after treatments and healthy volunteers.Conclusion: ACO patients from AE group have relatively lower eosinophil proportion and NK1R+ expression population eosinophil proportion, but higher plasma levels of cytokines and chemokines. Inhibitors of cytokines and chemokines are likely useful agents for treatment of ACO patients which had acute exacerbations.

scholarly journals A New MAP Kinase Protein Involved in Estradiol-Stimulated Reproduction of the Helminth ParasiteTaenia crassiceps

2010 ◽  
Vol 2010 ◽  
pp. 1-11 ◽  
Author(s):  
Galileo Escobedo ◽  
Gloria Soldevila ◽  
Guadalupe Ortega-Pierres ◽  
Jesús Ramsés Chávez-Ríos ◽  
Karen Nava ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Map Kinases ◽  
Host Cells ◽  
Cross Contamination ◽  
Flow Cytometry Analysis ◽  
Cellular Processes ◽  
17Β Estradiol ◽  
Activation Motif

MAP kinases (MAPK) are involved in the regulation of cellular processes such as reproduction and growth. In parasites, the role of MAPK has been scarcely studied. Here, we describe the participation of an ERK-like protein in estrogen-dependent reproduction of the helminth parasiteTaenia crassiceps. Our results show that 17β-estradiol induces a concentration-dependent increase in the bud number of in vitro cultured cysticerci. If parasites are also incubated in presence of an ERK-inhibitor, the stimulatory effect of estrogen is blocked. The expression of ERK-like mRNA and its corresponding protein was detected in the parasite. The ERK-like protein was over-expressed by all treatments. Nevertheless, a strong induction of phosphorylation of this protein was observed only in response to 17β-estradiol. Cross-contamination by host cells was discarded by flow cytometry analysis. Parasite cells expressing the ERK-like protein were exclusively located at the subtegument tissue by confocal microscopy. Finally, the ERK-like protein was separated by bidimensional electrophoresis and then sequenced, showing the conserved TEY activation motif, typical of all known ERK 1/2 proteins. Our results show that an ERK-like protein is involved in the molecular signalling during the interaction between the host andT. crassiceps, and may be considered as target for anti-helminth drugs design.

scholarly journals The role of dermis resident macrophages and their interaction with neutrophils in the early establishment of Leishmania major infection transmitted by sand fly bite

2020 ◽  
Vol 16 (11) ◽  
pp. e1008674
Author(s):  
Mariana M. Chaves ◽  
Sang Hun Lee ◽  
Olena Kamenyeva ◽  
Kashinath Ghosh ◽  
Nathan C. Peters ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Tyrosine Kinases ◽  
Leishmania Major ◽  
Direct Evidence ◽  
Sand Fly ◽  
Flow Cytometry Analysis ◽  
Cell Type ◽  
Severe Pathology ◽  
Early Establishment

There is substantial experimental evidence to indicate that Leishmania infections that are transmitted naturally by the bites of infected sand flies differ in fundamental ways from those initiated by needle inocula. We have used flow cytometry and intravital microscopy (IVM) to reveal the heterogeneity of sand fly transmission sites with respect to the subsets of phagocytes in the skin that harbor L. major within the first hours and days after infection. By flow cytometry analysis, dermis resident macrophages (TRMs) were on average the predominant infected cell type at 1 hr and 24 hr. By confocal IVM, the co-localization of L. major and neutrophils varied depending on the proximity of deposited parasites to the presumed site of vascular damage, defined by the highly localized swarming of neutrophils. Some of the dermal TRMs could be visualized acquiring their infections via transfer from or efferocytosis of parasitized neutrophils, providing direct evidence for the “Trojan Horse” model. The role of neutrophil engulfment by dermal TRMs and the involvement of the Tyro3/Axl/Mertk family of receptor tyrosine kinases in these interactions and in sustaining the anti-inflammatory program of dermal TRMs was supported by the effects observed in neutrophil depleted and in Axl-/-Mertk-/- mice. The Axl-/-Mertk-/- mice also displayed reduced parasite burdens but more severe pathology following L. major infection transmitted by sand fly bite.

scholarly journals Luteolin Promotes Cell Apoptosis by Inducing Autophagy in Hepatocellular Carcinoma

2017 ◽  
Vol 43 (5) ◽  
pp. 1803-1812 ◽  
Author(s):  
Zhijia Cao ◽  
Huainian Zhang ◽  
Xiaoyan Cai ◽  
Wei Fang ◽  
Dong Chai ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Flow Cytometry ◽  
Western Blotting ◽  
Cell Apoptosis ◽  
Flow Cytometry Analysis ◽  
Hoechst 33342 ◽  
Qrt Pcr ◽  
Induced Apoptosis ◽  
Luteolin Treatment

Background/Aims: Hepatocellular carcinoma (HCC) is the most common primary liver malignancy and is a leading cause of cancer-related death worldwide. Luteolin, a flavonoid from traditional Chinese medicine, shows anti-cancer activity in many cancer cells, including HCC. However, the mechanism underlying the action of luteolin in HCC, especially its role in regulating cell autophagy, remains unclear. In the present study, we investigated the role of luteolin in regulating cell autophagy and the role of autophagy in luteolin-induced apoptosis. Methods: The 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay (MTT) was used to investigate cell viability. Flow cytometry analysis was used to detect the cell cycle and cell apoptosis. Hoechst 33342 staining was used to detect cell apoptosis. Transmission electron microscopy was used to investigate autophagy. qRT-PCR and western blotting were used to detect apoptosis- and autophagy-related mRNAs and proteins. Results: Luteolin reduced the viability of SMMC-7721 cells in a time and dose-dependent manner, and induced significant G0/G1-phase arrest. In addition, the results of flow cytometry analysis and Hoechst 33342 staining showed that luteolin treatment increased the number of apoptotic cells obviously, and the results of qRT-PCR and western blotting showed that luteolin treatment increased caspase 8 and decreased bcl-2 at the mRNA and protein levels. Furthermore, luteolin increased the number of intracellular autophagosomes, promoted LC3B-I conversion to LC3B-II, and increased Beclin 1 expression. Finally, co-treatment with the autophagy inhibitor chloroquine weakened the effects of luteolin on cell apoptosis. Conclusion: Luteolin induced apoptosis in human liver cancer SMMC-7721 cells, partially via autophagy. Thus, luteolin could be used as a regulator of autophagy in HCC treatment.

scholarly journals Increased Lipid Peroxidation May Be Linked to Ferritin Levels Elevation in Adult-Onset Still’s Disease

Biomedicines ◽  
2021 ◽  
Vol 9 (11) ◽  
pp. 1508
Author(s):  
Po-Ku Chen ◽  
Kai-Jieh Yeo ◽  
Po-Hao Huang ◽  
Shih-Hsin Chang ◽  
Ching-Kun Chang ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Flow Cytometry ◽  
Fluorescence Intensity ◽  
Plasma Levels ◽  
Mean Fluorescence Intensity ◽  
Inflammatory Responses ◽  
Flow Cytometry Analysis ◽  
Inflammatory Parameters ◽  
Healthy Control ◽  
The Relationship

Lipid peroxidation (LPO) and hyper-ferritinemia are involved in inflammatory responses. Although hyper-ferritinemia is a characteristic of AOSD, its link to LPO remains unclear. We investigated the association between LPO and ferritin expression, and evaluated the relationship between LPO-related metabolites and inflammatory parameters. Mean fluorescence intensity (MFI) of LPO (C11-Biodipy581/591)-expressing PBMCs/monocytes in AOSD patients and healthy control (HC) subjects was determined by flow-cytometry analysis. Expression of ferritin and cytokines on PBMCs/macrophages was examined by immunoblotting. Plasma levels of LPO-related metabolites and cytokines were determined by ELISA and the MULTIPLEX platform, respectively. LPO MFI on PBMCs/monocytes were significantly higher in patients (median 4456 and 9091, respectively) compared with HC (1900, p < 0.05, and 4551, p < 0.01, respectively). Patients had higher ferritin expression on PBMCs (mean fold, 1.02) than HC (0.55, p < 0.05). Their ferritin expression levels on PBMCs stimulated with LPO inducers erastin or RSL3 (2.47 or 1.61, respectively) were higher than HC (0.84, p < 0.05, or 0.74, p < 0.01). Ferritin expression on erastin-treated/IL-1β-treated macrophages from patients were higher than those from HC (p < 0.001). The elevated levels of LPO-related metabolites, including malondialdehyde and 4-hydroxyalkenals, were positively correlated with disease activity scores, suggesting LPO involvement in AOSD pathogenesis. Increased ferritin expression on PBMCs/macrophages stimulated with LPO inducers indicates a link between LPO and elevated ferritin.

scholarly journals The Role of Moringa oleifera- Ifalmin® Formulation in Regulation of B220+IgM+ and B220+IgG+ in Diabetic Mice

2020 ◽  
Vol 10 (1) ◽  
pp. 43-48
Author(s):  
Wahyu Isnia Adharini ◽  
◽  
Ruri Vivian Nilamsari ◽  
Noviana Dwi Lestari ◽  
Nashi Widodo ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Flow Cytometry ◽  
Humoral Immunity ◽  
Moringa Oleifera ◽  
Normal Group ◽  
Metformin Treatment ◽  
Diabetic Mice ◽  
Flow Cytometry Analysis ◽  
Immunoglobulin Levels

It has been known that the immunoglobulin levels were altered in diabetes mellitus (DM) conditions. This study aimed to evaluate the levels of immunoglobulins in DM mice after the administration of Moringa oleifera-Ifalmin® formulation (MI). Streptozotocin, at a dose of 145 mg.kg-1, was injected intraperitoneally to experimental mice to obtain diabetic mice. The groups were divided into normal mice, diabetic mice without treatment, diabetic mice with metformin treatment (307.5 mg.kg-1 BW), and diabetic mice with MI treatment at dose 1 (M:I= 800 mg.kg-1 BW: 800 mg.kg-1 BW), dose 2 (M:I= 615 mg.kg-1 BW: 615 mg.kg-1 BW), and dose 3 (M:I= 800 mg.kg-1 BW: 615 mg.kg-1 BW). Mice were orally treated by MI for 14 days. Subsequently, the levels of immunoglobulin IgM and IgG were evaluated using flow cytometry analysis. IgM and IgG levels were significantly lower in the DM group than the normal group. These results indicated that DM altered immunoglobulin levels. MI treatment for 14 days significantly increased the number of IgM and IgG at the level equivalent to the normal group and significantly different as compared to the DM group. Based on the results, MI can be used as an immunomodulatory agent in humoral immunity through the precise regulation of IgM and IgG.

scholarly journals Enhancement of Antigen-specific functional responses by neutrophils from allergic patients.

1996 ◽  
Vol 183 (6) ◽  
pp. 2571-2579 ◽  
Author(s):  
J Monteseirín ◽  
M J Camacho ◽  
R Montaño ◽  
E Llamas ◽  
M Conde ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Surface ◽  
Respiratory Burst ◽  
Functional Role ◽  
Specific Binding ◽  
Flow Cytometry Analysis ◽  
Functional Responses ◽  
E Proteins ◽  
Healthy Donors

It has been demonstrated that neutrophils from healthy donors or from patients with inflammatory disorders can bind immunoglobulin (Ig) E proteins through binding to Mac-2/epsilon bp. Functional responses to allergens were assessed by measuring the respiratory burst and intracellular Ca2+ levels, and binding of allergens to neutrophils was assessed by flow cytometry analysis and fluorescence microscopy. In this article, we demonstrate that neutrophils sensitized to specific allergens (from allergic patients), but not from healthy donors, are sensitive to allergens of the same type as those that produce clinical allergic symptoms. The activation of neutrophils was analyzed by the induction of a respiratory burst that was detected with luminol-dependent chemiluminescence. Intracellular Ca2+ levels increased parallel to those of the inducing allergens. In addition, the specific binding of allergens on the cell surface was revealed by flow cytometry and allergen-FITC-labeled staining analyses. The present data suggest a restricted recognition of allergen by sensitive neutrophils, probably associated with the specific binding of the allergen to its corresponding IgE molecule, which is bound to the Mac-2/epsilon bp structure. These findings demonstrate a functional role of allergen-associated neutrophils during the allergic state.

Flow cytometry analysis reveals different activation profiles of thrombin- or TRAP-stimulated platelets in db/db mice. The regulatory role of PAR-3

2017 ◽  
Vol 65 ◽  
pp. 16-22 ◽  
Author(s):  
Hassan Kassassir ◽  
Karolina Siewiera ◽  
Marcin Talar ◽  
Tomasz Przygodzki ◽  
Cezary Watala
Keyword(s):  
Flow Cytometry ◽  
Regulatory Role ◽  
Flow Cytometry Analysis

scholarly journals A20 regulates the therapeutic effect of anti-PD-1 immunotherapy in melanoma

2020 ◽  
Vol 8 (2) ◽  
pp. e001866
Author(s):  
Weinan Guo ◽  
Jinyuan Ma ◽  
Sen Guo ◽  
Huina Wang ◽  
Sijia Wang ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Mouse Model ◽  
Therapeutic Effect ◽  
Immune Checkpoint ◽  
Antitumor Immunity ◽  
Flow Cytometry Analysis ◽  
Melanoma Patients

BackgroundThe therapeutic effect of immune checkpoint blockers, especially the neutralizing antibodies of programmed cell death (PD-1) and its ligand programmed death ligand 1 (PD-L1), has been well verified in melanoma. Nevertheless, the dissatisfactory response rate and the occurrence of resistance significantly hinder the treatment effect. Inflammation-related molecules like A20 are greatly implicated in cancer immune response, but the role of tumorous A20 in antitumor immunity and immunotherapy efficacy remains elusive.MethodsThe association between tumorous A20 expression and the effect of anti-PD-1 immunotherapy was determined by immunoblotting, immunofluorescence staining and flow cytometry analysis of primary tumor specimens from melanoma patients. Preclinical mouse model, in vitro coculture system, immunohistochemical staining and flow cytometry analysis were employed to investigate the role of A20 in regulating the effect of anti-PD-1 immunotherapy. Bioinformatics, mass spectrum analysis and a set of biochemical analyzes were used to figure out the underlying mechanism.ResultsWe first discovered that upregulated A20 was associated with impaired antitumor capacity of CD8+T cells and poor response to anti-PD-1 immunotherapy in melanoma patients. Subsequent functional studies in preclinical mouse model and in vitro coculture system proved that targeting tumorous A20 prominently improved the effect of immunotherapy through the invigoration of infiltrating CD8+T cells via the regulation of PD-L1. Mechanistically, A20 facilitated the ubiquitination and degradation of prohibitin to potentiate STAT3 activation and PD-L1 expression. Moreover, tumorous A20 expression was highly associated with the ratio of Ki-67 percentage in circulating PD-1+CD8+T cells to tumor burden.ConclusionsTogether, our findings uncover a novel crosstalk between inflammatory molecules and antitumor immunity in melanoma, and highlight that A20 can be exploited as a promising target to bring clinical benefit to melanomas refractory to immune checkpoint blockade.

scholarly journals Spectrum of acute leukemias diagnosed on flow cytometry: Analysis from tertiary care centre from North India

2015 ◽  
Vol 1 (1) ◽  
pp. 12-15 ◽  
Author(s):  
Surendra Koju ◽  
Man Updesh Singh Sachdeva ◽  
Praveen Bose ◽  
Neelam Varma
Keyword(s):  
Flow Cytometry ◽  
Tertiary Care ◽  
Antigen Expression ◽  
North India ◽  
Molecular Characteristics ◽  
Flow Cytometry Analysis ◽  
Tertiary Care Centre ◽  
Acute Leukemias

BACKGROUND: Acute leukemias (ALs) are a heterogeneous group of malignancies with varying clinical, morphologic, immunologic, and molecular characteristics. WHO 2008 classification of ALs require a multi-parametric approach to the diagnosis. This study aims to evaluate the role of flow cytometry in diagnosis and sub-classification of acute leukemias. METHODS: Consecutive patients of adult and paediatric ALs during June 2012 to May 2013 were retrospectively analyzed and studied using BD FACS Canto-II flow cytometer. The results of immunophenotyping were reviewed and analyzed for cross-lineage antigen expression. RESULT: Over a period of year, 422 individuals were diagnosed as AL. There were 287 males and 135 females with M:F = 2.1:1. There were 237 adults & 185 children. 36.3% were AML and 60.4% were ALL, while 3.3% of cases were mixed phenotypic acute leukemia (MPAL). The commonest WHO subtype in AML group was AML with maturation being 31%. In case of ALL there were 83.9% B-ALLs and 16.1% T-ALLs. In MPAL B-Myeloid was 71.4%, whereas T-Myeloid was 28.6% of cases. Both AML and MPAL were more frequently seen in adults accounting to 83% and 92.9% respectively of all ALs cases. In contrast, 62% of ALLs were children and only 38% were adults. Out of all ALs, 37.6% of showed cross lineage antigen expression. In AML, B-ALL and T-ALL cross lineage antigen expression were 26.14%, 39.71% and 82.9% respectively. CONCLUSION: Flow cytometry is useful in diagnosis and sub classification of AL. It is essential in cytochemical myeloperoxidase (MPO) negative cases. Cross- lineage antigen expression is frequent in ALs, and hence, lineage specific intra-cytoplasmic antibodies including anti-MPO and cytoplasmic-CD3 are essential for correct categorization of ALs. DOI: http://dx.doi.org/10.3126/acclm.v1i1.12308 Ann. Clin. Chem. & Lab. Med. 1(1) 2015: 12-15

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