Macrophage supernatant infected with <em>Leishmania major</em> mediates the cytology of fibroblast cells in skin wounds

Objective: Healing of Cutaneous Leishmaniasis relies on the effective and modulates protective immune responses. Although the immune system is necessary to eliminate the parasite, it could be considered as the main cause of ulcers. Therefore, main aim of this study was to explore the possible regulatory functions of macrophage supernatant infected with Leishmania major on the fibroblast cells. Materials and Methods: In this experimental study, different concentrations of infected macrophage supernatant extract (50, 100, 150, 200, and 250μg/mL) were tested at different times (6, 24, 48, and 72h) and the effect of the leishmanicidal extract on fibroblast cells was determined by MTS assay. Also, the flow-cytometry technique was used for the investigation of apoptosis induction percentage. Results: MTS assay showed that the leishmanicidal effect of infected macrophage supernatant extract was dependent on the concentration and the time of treatment. So, the best efficacy was observed in 200 μg/mL with 72 hours exposure time. Flow cytometry analysis showed that the infected macrophage supernatant extract could induce apoptosis in cultured fibroblasts. Conclusions: We have demonstrated that reduction of survival rate and induction of apoptosis in fibroblasts displayed a similar manner to keratinocytes when exposed to infected macrophages with L. major. Our data suggest that such a phenomenon can be the underlying cause of lesions with scarring, and future, the mechanism remains to be elucidated.

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

PLoS Pathogens ◽

10.1371/journal.ppat.1008674 ◽

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.

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Real-time flow cytometry analysis of permeability transition in isolated mitochondria

Experimental Cell Research ◽

10.1016/j.yexcr.2003.10.030 ◽

2004 ◽

Vol 294 (1) ◽

pp. 106-117 ◽

Cited By ~ 55

Author(s):

Hervé Lecoeur ◽

Alain Langonné ◽

Ludwig Baux ◽

Dominique Rebouillat ◽

Pierre Rustin ◽

...

Keyword(s):

Flow Cytometry ◽

Real Time ◽

Permeability Transition ◽

Flow Cytometry Analysis ◽

Isolated Mitochondria ◽

Time Flow

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Flow Cytometry Analysis of the Reactive Oxygen Species in Immature Granulocytes in Septic Patient

Case Medical Research ◽

10.31525/ct1-nct03846596 ◽

2000 ◽

Author(s):

Keyword(s):

Reactive Oxygen Species ◽

Flow Cytometry ◽

Septic Patient ◽

Reactive Oxygen ◽

Oxygen Species ◽

Flow Cytometry Analysis

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In Vitro Evaluation of Chitosan Induced Cytotoxicity against Cancerous Cell lines: MCF-7, Saos-2 and HeLa

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520619666190507113949 ◽

2019 ◽

Vol 19 ◽

Author(s):

Zeinab Abedian ◽

Niloofar Jenabian ◽

Ali Akbar Moghadamnia ◽

Ebrahim Zabihi ◽

Roghayeh Pourbagher ◽

...

Keyword(s):

Flow Cytometry ◽

Cell Lines ◽

Anticancer Agents ◽

Fibroblast Cells ◽

Apoptosis Induction ◽

Cytotoxic Effects ◽

Cancerous Cell ◽

Significant Concentration ◽

Mcf 7

Objective/ Background: Cancer is still the most common cause of morbidity in world and new powerful anticancer agents without severe side effects from natural sources is important. Methods: The evaluation of cytotoxicity and apoptosis induction was carried out in MCF-7,HeLa and Saos-2 as cancerous cell lines with different histological origin and human fibroblast served as control normal cell. The cells were treated with different concentrations of chitosan and the cytotoxicity was determined using MTT assay after 24, 48 and 72 h .The mode of death was evaluated by flow cytometry . Results: While both types of chitosan showed significant concentration-dependently cytotoxic effects against the three cancerous cell lines, fibroblast cells showed somehow more compatibility with chitosan. On the other hand, there were no significant differences between LMWC and HMWC cytotoxicity in all cell lines. The flow cytometry results showed the apoptosis pattern of death more in Saos-2 and HeLa while necrosis was more observable with MCF7. Also higher viability with both types of chitosan was seen in fibroblast as normal cells Conclusion: Chitosan shows anticancerous effect against 3 cancerous cell lines, while it is compatible with normal diploid fibroblast cells. Furthermore, it seems that the molecular weight of chitosan does not affect its anticancerous property.

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Evaluating the Effect of Cinnarizine on Promastigotes and Amastigotes forms of Leishmania major

Infectious Disorders - Drug Targets ◽

10.2174/1871526518666181113114820 ◽

2020 ◽

Vol 20 (4) ◽

pp. 550-555 ◽

Cited By ~ 1

Author(s):

Lima Asgharpour Sarouey ◽

Parvaneh Rahimi-Moghaddam ◽

Fatemeh Tabatabaie ◽

Khadijeh Khanaliha

Keyword(s):

Flow Cytometry ◽

Cutaneous Leishmaniasis ◽

Leishmania Major ◽

Inhibitory Effect ◽

Control Group ◽

Secondary Infections ◽

Ic50 Values ◽

J774 Cells ◽

Mtt Assays

: As an important global disease, cutaneous leishmaniasis is associated with complications such as secondary infections and atrophic scars. The first line treatment with antimonials is expensive and reported to have serious side effects and enhance resistance development. The main objective of this study was to evaluate the effect of Cinnarizine on standard strains of Leishmania major because of paucity of information on this subject. Methods: In this experimental study, four concentrations of the drug (5, 10, 15 and 20 μg/ml) were added to Leishmania major cultures at 24, 48 and 72 hours intervals. MTT assays were performed to determine parasite viability and drug toxicity. Leishmania major promastigotes were augmented to the in vitro cultured macrophages (J774 cells) and then incubated for 72 hours. Half maximal inhibitory concentration (IC50) was ascertained by counting parasites. The inhibitory effect of the drug was compared with that of Glucantime. Flow-cytometry was performed to investigate apoptosis. Each test was repeated thrice. Results: The IC50 values of Cinnarizine after 72 hours were calculated to be 34.76 μg/ml and 23.73 μg/ml for promastigotes and amastigotes, respectively. The results of MTT assays showed 48 % promastigote viability after 72 hour-exposure to Cinnarizine at 20 μg/ml concentration. Programmed cell death in promastigote- and amastigote-infected macrophages was quantified to be 13.66 % and 98.7 %, respectively. Flow- cytometry analysis indicated that Cinnarizine induced early and late apoptosis in parasites. All treatments produced results which differed significantly from control group (P<0.05). Conclusion: Cinnarizine showed low toxicity with anti-leishmanial and apoptosis effects on both promastigote and intracellular amastigote forms. Therefore, we may suggest further assessment on animal models of this drug as candidates for cutaneous leishmaniasis therapy.

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A Synergic Potential of Antimicrobial Peptides against Pseudomonas syringae pv. actinidiae

Molecules ◽

10.3390/molecules26051461 ◽

2021 ◽

Vol 26 (5) ◽

pp. 1461

Author(s):

Nuno Mariz-Ponte ◽

Laura Regalado ◽

Emil Gimranov ◽

Natália Tassi ◽

Luísa Moura ◽

...

Keyword(s):

Flow Cytometry ◽

Antimicrobial Peptides ◽

Pseudomonas Syringae ◽

Pathogenic Bacteria ◽

High Efficiency ◽

Bacterial Canker ◽

Membrane Permeation ◽

Flow Cytometry Analysis ◽

Pathogenic Agent ◽

Peptide Mixtures

Pseudomonas syringae pv. actinidiae (Psa) is the pathogenic agent responsible for the bacterial canker of kiwifruit (BCK) leading to major losses in kiwifruit productions. No effective treatments and measures have yet been found to control this disease. Despite antimicrobial peptides (AMPs) having been successfully used for the control of several pathogenic bacteria, few studies have focused on the use of AMPs against Psa. In this study, the potential of six AMPs (BP100, RW-BP100, CA-M, 3.1, D4E1, and Dhvar-5) to control Psa was investigated. The minimal inhibitory and bactericidal concentrations (MIC and MBC) were determined and membrane damaging capacity was evaluated by flow cytometry analysis. Among the tested AMPs, the higher inhibitory and bactericidal capacity was observed for BP100 and CA-M with MIC of 3.4 and 3.4–6.2 µM, respectively and MBC 3.4–10 µM for both. Flow cytometry assays suggested a faster membrane permeation for peptide 3.1, in comparison with the other AMPs studied. Peptide mixtures were also tested, disclosing the high efficiency of BP100:3.1 at low concentration to reduce Psa viability. These results highlight the potential interest of AMP mixtures against Psa, and 3.1 as an antimicrobial molecule that can improve other treatments in synergic action.

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