scholarly journals Endochin-like quinolone-300 and ELQ-316 inhibit Babesia bovis, B. bigemina, B. caballi and Theileria equi

2020 ◽  
Author(s):  
Carlos Suarez ◽  
Marta G. Silva ◽  
Reginaldo G. Bastos ◽  
J. Stone Doggett ◽  
Michael K. Riscoe ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Babesia Bovis ◽  
Host Cells ◽  
Bovine Babesiosis ◽  
Theileria Equi ◽  
Peripheral Blood Mononuclear ◽  
Equine Piroplasmosis ◽  
Apicomplexan Parasites ◽  
And Control

Abstract Background: The most common apicomplexan parasites causing bovine babesiosis are Babesia bovis and B. bigemina, while B. caballi and Theileria equi are responsible for equine piroplasmosis. Treatment and control of these diseases are usually achieved using potentially toxic chemotherapeutics, such as imidocarb diproprionate, but drug-resistant parasites are emerging, and alternative effective and safer drugs are needed. Endochin-like quinolones (ELQ)-300 and ELQ-316 proved safe and efficacious against related apicomplexans, such as Plasmodium spp., and ELQ-316 was also effective against B. microti, without showing toxicity in mammals.Methods: Inhibitory effects of ELQ-300 and ELQ-316 were assessed on the growth of cultured B. bovis, B. bigemina, B. caballi and T. equi. Percentage of parasitized erythrocytes was measured by flow cytometry. Effect of the ELQ drugs on the viability of actively replicating horse and bovine peripheral blood mononuclear cells (PBMC) was assessed by ELISA. Results: We calculated IC50 ranging from 0.04 to 0.37 nM for ELQ-300, and from 0.002 to 0.1 nM for ELQ-316 at 72 hr among all cultured parasites tested. None of the parasites tested were able to replicate in cultures in the presence of the ELQs-300 and ELQ-316 at IC100, which range from 1.3 to 5.7 nM for ELQ-300 and from 1.0 to 6.0 nM for ELQ-316 at 72 hours. Neither ELQ-300 nor ELQ-316 altered the viability of equine and bovine PBMC at their IC100 in in vitro testing. Conclusions: ELQ-300 and ELQ-316 have a significant inhibitory activity on the main parasites responsible for bovine babesiosis and equine piroplasmosis at doses that are tolerable to host cells. These ELQ drugs may be viable candidates for developing alternative protocols for the treatment of bovine babesiosis and equine piroplasmosis.

scholarly journals Endochin-like quinolone-300 and ELQ-316 inhibit Babesia bovis, B. bigemina, B. caballi and Theileria equi

2020 ◽  
Author(s):  
Marta G. Silva ◽  
Reginaldo G. Bastos ◽  
J. Stone Doggett ◽  
Michael K. Riscoe ◽  
Sovitj Pou ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Colorimetric Assay ◽  
Babesia Bovis ◽  
Host Cells ◽  
Bovine Babesiosis ◽  
Theileria Equi ◽  
Peripheral Blood Mononuclear ◽  
Equine Piroplasmosis ◽  
And Control

Abstract Background: The most common apicomplexan parasites causing bovine babesiosis are Babesia bovis and B. bigemina, while B. caballi and Theileria equi are responsible for equine piroplasmosis. Treatment and control of these diseases are usually achieved using potentially toxic chemotherapeutics, such as imidocarb diproprionate, but drug-resistant parasites are emerging, and alternative effective and safer drugs are needed. Endochin-like quinolones (ELQ)-300 and ELQ-316 proved safe and efficacious against related apicomplexans, such as Plasmodium spp., and ELQ-316 was also effective against B. microti, without showing toxicity in mammals.Methods: Inhibitory effects of ELQ-300 and ELQ-316 were assessed on the growth of cultured B. bovis, B. bigemina, B. caballi and T. equi. Percentage of parasitized erythrocytes was measured by flow cytometry. Effect of the ELQ drugs on the viability of horse and bovine peripheral blood mononuclear cells (PBMC) was assessed by monitoring cell metabolic activity using a colorimetric assay.Results: We calculated IC50 ranging from 0.04 to 0.37 nM for ELQ-300, and from 0.002 to 0.1 nM for ELQ-316 at 72 h among all cultured parasites tested. None of the parasites tested were able to replicate in cultures in the presence of the ELQ-300 and ELQ-316 at IC100, which range from 1.3 to 5.7 nM for ELQ-300 and from 1.0 to 6.0 nM for ELQ-316 at 72 h. Neither ELQ-300 nor ELQ-316 altered the viability of equine and bovine PBMC at their IC100 in in vitro testing. Conclusions: ELQ-300 and ELQ-316 showed significant inhibitory activity on the main parasites responsible for bovine babesiosis and equine piroplasmosis at doses that are tolerable to host cells. These ELQ drugs may be viable candidates for developing alternative protocols for the treatment of bovine babesiosis and equine piroplasmosis.

scholarly journals Endochin-like quinolone-300 and ELQ-316 inhibit Babesia bovis, B. bigemina, B. caballi and Theileria equi 

2020 ◽  
Author(s):  
Marta G. Silva ◽  
Reginaldo G. Bastos ◽  
J. Stone Doggett ◽  
Michael K. Riscoe ◽  
Sovitj Pou ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Colorimetric Assay ◽  
Babesia Bovis ◽  
Host Cells ◽  
Bovine Babesiosis ◽  
Theileria Equi ◽  
Peripheral Blood Mononuclear ◽  
Equine Piroplasmosis ◽  
And Control

Abstract Background: The most common apicomplexan parasites causing bovine babesiosis are Babesia bovis and B. bigemina, while B. caballi and Theileria equi are responsible for equine piroplasmosis. Treatment and control of these diseases are usually achieved using potentially toxic chemotherapeutics, such as imidocarb diproprionate, but drug-resistant parasites are emerging, and alternative effective and safer drugs are needed. Endochin-like quinolones (ELQ)-300 and ELQ-316 proved safe and efficacious against related apicomplexans, such as Plasmodium spp., and ELQ-316 was also effective against B. microti, without showing toxicity in mammals.Methods: Inhibitory effects of ELQ-300 and ELQ-316 were assessed on the growth of cultured B. bovis, B. bigemina, B. caballi and T. equi. Percentage of parasitized erythrocytes was measured by flow cytometry. Effect of the ELQ drugs on the viability of horse and bovine peripheral blood mononuclear cells (PBMC) was assessed by monitoring cell metabolic activity using a colorimetric assay.Results: We calculated IC50 ranging from 0.04 to 0.37 nM for ELQ-300, and from 0.002 to 0.1 nM for ELQ-316 at 72 h among all cultured parasites tested. None of the parasites tested were able to replicate in cultures in the presence of the ELQ-300 and ELQ-316 at IC100, which range from 1.3 to 5.7 nM for ELQ-300 and from 1.0 to 6.0 nM for ELQ-316 at 72 h. Neither ELQ-300 nor ELQ-316 altered the viability of equine and bovine PBMC at their IC100 in in vitro testing. Conclusions: ELQ-300 and ELQ-316 showed significant inhibitory activity on the main parasites responsible for bovine babesiosis and equine piroplasmosis at doses that are tolerable to host cells. These ELQ drugs may be viable candidates for developing alternative protocols for the treatment of bovine babesiosis and equine piroplasmosis.

scholarly journals Endochin-like quinolone-300 and ELQ-316 inhibit Babesia bovis, B. bigemina, B. caballi and Theileria equi

2020 ◽  
Author(s):  
Marta G. Silva ◽  
Reginaldo G. Bastos ◽  
J. Stone Doggett ◽  
Michael K. Riscoe ◽  
Sovitj Pou ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Colorimetric Assay ◽  
Babesia Bovis ◽  
Host Cells ◽  
Bovine Babesiosis ◽  
Theileria Equi ◽  
Peripheral Blood Mononuclear ◽  
Equine Piroplasmosis ◽  
And Control

Abstract Background: The most common apicomplexan parasites causing bovine babesiosis are Babesia bovis and B. bigemina, while B. caballi and Theileria equi are responsible for equine piroplasmosis. Treatment and control of these diseases are usually achieved using potentially toxic chemotherapeutics, such as imidocarb diproprionate, but drug-resistant parasites are emerging, and alternative effective and safer drugs are needed. Endochin-like quinolones (ELQ)-300 and ELQ-316 proved safe and efficacious against related apicomplexans, such as Plasmodium spp., and ELQ-316 was also effective against B. microti, without showing toxicity in mammals.Methods: Inhibitory effects of ELQ-300 and ELQ-316 were assessed on the growth of cultured B. bovis, B. bigemina, B. caballi and T. equi. Percentage of parasitized erythrocytes was measured by flow cytometry. Effect of the ELQ drugs on the viability of horse and bovine peripheral blood mononuclear cells (PBMC) was assessed by monitoring cell metabolic activity using a colorimetric assay.Results: We calculated IC50 ranging from 0.04 to 0.37 nM for ELQ-300, and from 0.002 to 0.1 nM for ELQ-316 at 72 h among all cultured parasites tested. None of the parasites tested were able to replicate in cultures in the presence of the ELQ-300 and ELQ-316 at IC100, which range from 1.3 to 5.7 nM for ELQ-300 and from 1.0 to 6.0 nM for ELQ-316 at 72 h. Neither ELQ-300 nor ELQ-316 altered the viability of equine and bovine PBMC at their IC100 in in vitro testing. Conclusions: ELQ-300 and ELQ-316 showed significant inhibitory activity on the main parasites responsible for bovine babesiosis and equine piroplasmosis at doses that are tolerable to host cells. These ELQ drugs may be viable candidates for developing alternative protocols for the treatment of bovine babesiosis and equine piroplasmosis.

scholarly journals Endochin-like quinolone-300 and ELQ-316 inhibit Babesia bovis, B. bigemina, B. caballi and Theileria equi

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Marta G. Silva ◽  
Reginaldo G. Bastos ◽  
J. Stone Doggett ◽  
Michael K. Riscoe ◽  
Sovitj Pou ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Inhibitory Concentration ◽  
Colorimetric Assay ◽  
Babesia Bovis ◽  
Host Cells ◽  
Babesia Microti ◽  
Bovine Babesiosis ◽  
Theileria Equi ◽  
Peripheral Blood Mononuclear ◽  
Equine Piroplasmosis

Abstract Background The most common apicomplexan parasites causing bovine babesiosis are Babesia bovis and B. bigemina, while B. caballi and Theileria equi are responsible for equine piroplasmosis. Treatment and control of these diseases are usually achieved using potentially toxic chemotherapeutics, such as imidocarb diproprionate, but drug-resistant parasites are emerging, and alternative effective and safer drugs are needed. The endochin-like quinolones (ELQ)-300 and ELQ-316 have been proven to be safe and efficacious against related apicomplexans, such as Plasmodium spp., with ELQ-316 also being effective against Babesia microti, without showing toxicity in mammals. Methods The inhibitory effects of ELQ-300 and ELQ-316 were assessed on the growth of cultured B. bovis, B. bigemina, B. caballi and T. equi. The percentage of parasitized erythrocytes was measured by flow cytometry, and the effect of the ELQ compounds on the viability of horse and bovine peripheral blood mononuclear cells (PBMC) was assessed by monitoring cell metabolic activity using a colorimetric assay. Results We calculated the half maximal inhibitory concentration (IC50) at 72 h, which ranged from 0.04 to 0.37 nM for ELQ-300, and from 0.002 to 0.1 nM for ELQ-316 among all cultured parasites tested at 72 h. None of the parasites tested were able to replicate in cultures in the presence of ELQ-300 and ELQ-316 at the maximal inhibitory concentration (IC100), which ranged from 1.3 to 5.7 nM for ELQ-300 and from 1.0 to 6.0 nM for ELQ-316 at 72 h. Neither ELQ-300 nor ELQ-316 altered the viability of equine and bovine PBMC at their IC100 in in vitro testing. Conclusions The compounds ELQ-300 and ELQ-316 showed significant inhibitory activity on the main parasites responsible for bovine babesiosis and equine piroplasmosis at doses that are tolerable to host cells. These ELQ drugs may be viable candidates for developing alternative protocols for the treatment of bovine babesiosis and equine piroplasmosis.

scholarly journals Long-term stable hematopoietic chimerism following marrow transplantation for acute lymphoblastic leukemia: a case report with in vitro marrow culture studies

Blood ◽  
1983 ◽  
Vol 62 (4) ◽  
pp. 869-872 ◽  
Author(s):  
JW Singer ◽  
A Keating ◽  
R Ramberg ◽  
R McGuffin ◽  
JE Sanders ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Mononuclear Cells ◽  
Lymphoblastic Leukemia ◽  
Host Cells ◽  
Marrow Graft ◽  
Peripheral Blood Mononuclear ◽  
Hematopoietic Chimerism

Abstract This article describes the course of a patient who received an allogeneic marrow graft from his HLA-identical sister for acute lymphoblastic leukemia in second remission. In the second month after grafting, marrow aspirates showed the presence of 7%-10% lymphoblasts. In addition, cytogenetic examination indicated the persistence of host cells. Thereafter, the patient had morphologically normal marrow examinations, with no evidence for recurrent leukemia. In addition, stable hematopoietic chimerism in both the lymphoid and myeloid cell lines has persisted for over 5 yr. Between 20% and 50% of phytohemagglutinin-stimulated peripheral blood mononuclear cells were host-derived on repeated studies. A marrow sample 4 yr after transplantation was established in long-term culture and produced 2% host granulocyte-macrophage colonies at its inception, but 24% host colonies by week 4. Despite this persistent chimerism, no in vitro or in vivo abnormalities of hematopoiesis have been detected.

scholarly journals Cell-in-cell phenomenon: leukocyte engulfment by non-tumorigenic cells and cancer cell lines

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Mareike F. Bauer ◽  
Michael Hader ◽  
Markus Hecht ◽  
Maike Büttner-Herold ◽  
Rainer Fietkau ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
Mononuclear Cells ◽  
Cancer Cell Lines ◽  
Host Cells ◽  
Target Cells ◽  
Peripheral Blood Mononuclear ◽  
The Impact ◽  
Microwave Hyperthermia

Abstract Background Research on cell-in-cell (CIC) phenomena, including entosis, emperipolesis and cannibalism, and their biological implications has increased in recent years. Homotypic and heterotypic engulfment of various target cells by numerous types of host cells has been studied in vitro and in tissue sections. This work has identified proteins involved in the mechanism and uncovered evidence for CIC as a potential histopathologic predictive and prognostic marker in cancer. Our experimental study focused on non-professional phagocytosis of leukocytes. Results We studied the engulfment of peripheral blood mononuclear cells isolated from healthy donors by counting CIC structures. Two non-tumorigenic cell lines (BEAS-2B, SBLF-9) and two tumour cell lines (BxPC3, ICNI) served as host cells. Immune cells were live-stained and either directly co-incubated or treated with irradiation or with conventional or microwave hyperthermia. Prior to co-incubation, we determined leukocyte viability for each batch via Annexin V-FITC/propidium iodide staining. All host cells engulfed their targets, with uptake rates ranging from 1.0% ± 0.5% in BxPC3 to 8.1% ± 5.0% in BEAS-2B. Engulfment rates of the cancer cell lines BxPC3 and ICNI (1.6% ± 0.2%) were similar to those of the primary fibroblasts SBLF-9 (1.4% ± 0.2%). We found a significant negative correlation between leukocyte viability and cell-in-cell formation rates. The engulfment rate rose when we increased the dose of radiotherapy and prolonged the impact time. Further, microwave hyperthermia induced higher leukocyte uptake than conventional hyperthermia. Using fluorescent immunocytochemistry to descriptively study the proteins involved, we detected ring-like formations of diverse proteins around the leukocytes, consisting, among others, of α-tubulin, integrin, myosin, F-actin, and vinculin. These results suggest the involvement of actomyosin contraction, cell-cell adhesion, and the α-tubulin cytoskeleton in the engulfment process. Conclusions Both non-tumorigenic and cancer cells can form heterotypic CIC structures by engulfing leukocytes. Decreased viability and changes caused by microwave and X-ray irradiation trigger non-professional phagocytosis.

Interferon-γ-mediated pathways and in vitro PBMC proliferation in HIV-infected patients

2009 ◽  
Vol 390 (2) ◽  
pp. 115-123 ◽  
Author(s):  
Katharina Schroecksnadel ◽  
Christiana Winkler ◽  
Ernst R. Werner ◽  
Mario Sarcletti ◽  
Nikolaus Romani ◽  
...  
Keyword(s):  
Proliferative Response ◽  
Mononuclear Cells ◽  
Interferon Γ ◽  
Pokeweed Mitogen ◽  
Peripheral Blood Mononuclear ◽  
Tryptophan Degradation ◽  
Infected Pbmcs ◽  
Ifn Γ ◽  
And Control

AbstractHIV infection is characterized by progressive immunodeficiency: HIV-infected peripheral blood mononuclear cells (PBMCs) cannot properly react to stimulation with allo-antigens and mitogens. In this study, we examined interferon-γ (IFN-γ)-mediated pathways and the proliferative response of mitogen-stimulated HIV-infected PBMCsin vitro. PBMCs of 30 HIV-infected patients were stimulated with the mitogens concanavalin A (Con A), phytohemagglutinin (PHA), and pokeweed mitogen (PWM). Mitogen stimulation induced expression of IFN-γ, GTP cyclohydrolase I (GCH-I), and indoleamine (2,3)-dioxygenase (IDO) resulting in enhanced neopterin formation and tryptophan degradation by HIV-infected and control PBMCs. IFN-γ concentrations correlated with neopterin levels and tryptophan degradation. Proliferative responses to PHA and PWM cytokine were lower in HIV patients, with IFN-γ formation predicting proliferative responses. Higher mRNA expression of IFN-γ, GCH-I and IDO after 6 h was related to better proliferative responses in HIV-infected PBMCs. In conclusion, induction of IFN-γ and subsequent enzymes appears to importantly influence the proliferative response of HIV-infected PBMCsin vitro, suggesting a prominent role of the cytokine in the development of immunodeficiency.

scholarly journals Long-term stable hematopoietic chimerism following marrow transplantation for acute lymphoblastic leukemia: a case report with in vitro marrow culture studies

Blood ◽  
1983 ◽  
Vol 62 (4) ◽  
pp. 869-872
Author(s):  
JW Singer ◽  
A Keating ◽  
R Ramberg ◽  
R McGuffin ◽  
JE Sanders ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Mononuclear Cells ◽  
Lymphoblastic Leukemia ◽  
Host Cells ◽  
Marrow Graft ◽  
Peripheral Blood Mononuclear ◽  
Hematopoietic Chimerism

This article describes the course of a patient who received an allogeneic marrow graft from his HLA-identical sister for acute lymphoblastic leukemia in second remission. In the second month after grafting, marrow aspirates showed the presence of 7%-10% lymphoblasts. In addition, cytogenetic examination indicated the persistence of host cells. Thereafter, the patient had morphologically normal marrow examinations, with no evidence for recurrent leukemia. In addition, stable hematopoietic chimerism in both the lymphoid and myeloid cell lines has persisted for over 5 yr. Between 20% and 50% of phytohemagglutinin-stimulated peripheral blood mononuclear cells were host-derived on repeated studies. A marrow sample 4 yr after transplantation was established in long-term culture and produced 2% host granulocyte-macrophage colonies at its inception, but 24% host colonies by week 4. Despite this persistent chimerism, no in vitro or in vivo abnormalities of hematopoiesis have been detected.

scholarly journals Role of Mycoplasma penetransEndonuclease P40 as a Potential Pathogenic Determinant

1999 ◽  
Vol 67 (9) ◽  
pp. 4456-4462 ◽  
Author(s):  
Mourad Bendjennat ◽  
Alain Blanchard ◽  
Mohammed Loutfi ◽  
Luc Montagnier ◽  
Elmostafa Bahraoui
Keyword(s):  
Mononuclear Cells ◽  
Morphological Changes ◽  
Fluorescein Isothiocyanate ◽  
Host Cells ◽  
Cell Surface Receptor ◽  
Dependent Manner ◽  
Peripheral Blood Mononuclear ◽  
Surface Receptor

ABSTRACT Recently, we reported the purification to homogeneity and characterization of Ca2+- and Mg2+-dependent endonuclease P40 produced by Mycoplasma penetrans (M. Bendjennat, A. Blanchard, M. Loutfi, L. Montagnier, and E. Bahraoui, J. Bacteriol. 179; 2210–2220, 1997), a mycoplasma which was isolated for the first time from the urine of human immunodeficiency virus-infected patients. To evaluate how this nuclease could interact with host cells, we tested its effect on CEM and Molt-4 lymphocytic cell lines and on peripheral blood mononuclear cells. We observed that 10−7to 10−9 M P40 is able to mediate a cytotoxic effect. We found that 100% of cells were killed after 24 h of incubation with 10−7 M P40 while only 40% cytotoxicity was obtained after 72 h of incubation with 10−9 M P40. Phase-contrast microscopy observations of P40-treated cells revealed morphological changes, including pronounced blebbing of the plasma membrane and cytoplasmic shrinkage characteristic of programmed cell death, which is in agreement with the internucleosomal fragmentation of P40-treated cell DNA as shown by agarose gel electrophoresis. We showed that 125I-radiolabeled or fluorescein isothiocyanate-labeled P40 was able to bind specifically in a dose-dependent manner to the cell membrane of CEM cells, which suggested that the cytotoxicity of P40 endonuclease was mediated by its interaction with the cell surface receptor(s). The concentration of unlabeled P40 required to inhibit by 50% the formation of125I-P40-CEM complexes was about 3 × 10−9 M, indicating a high-affinity interaction. Both P40 interaction and cytotoxicity are Ca2+ dependent. Our results suggest that the cytotoxicity of M. penetransobserved in vitro is mediated at least partially by secreted P40, which, after interaction with host cells, can induce an apoptosis-like death. These results strongly suggest a major role of mycoplasmal nucleases as potential pathogenic determinants.

scholarly journals Tetraoxanes as inhibitors of apicomplexan parasites Plasmodium falciparum and Toxoplasma gondii and anti-cancer molecules

2015 ◽  
Vol 80 (11) ◽  
pp. 1339-1359 ◽  
Author(s):  
Dejan Opsenica ◽  
Jelena Radivojevic ◽  
Ivana Matic ◽  
Tijana Stajner ◽  
Slavica Knezevic-Usaj ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Toxoplasma Gondii ◽  
Mononuclear Cells ◽  
Antimalarial Activity ◽  
Human Cancer ◽  
Subcutaneous Administration ◽  
Peripheral Blood Mononuclear ◽  
Normal Peripheral Blood ◽  
Apicomplexan Parasites

New cyclohexylidene 1,2,4,5-tetraoxanes with polar guanidine and urea based groups were synthesized and evaluated for antimalarial activity against chloroquine resistant and susceptible Plasmodium falciparum strains. Derivatives showed moderate nM range antimalarial activities and low cytotoxicity. N-phenyl-urea derivative 24 exhibited best resistant indices (RIW2 = 0.44, RITM91C235 = 0.80), and was not toxic against human normal peripheral blood mononuclear cells (IC50 > 200 ?M). Seven derivatives were tested in vitro against four human cancer cell lines and they demonstrated high selectivity toward leukemia K562 cells. One compound, derivative 21 with a primary amino-group, was the first tetraoxane tested in vivo against Toxoplasma gondii as another Apicomplexan parasite. Subcutaneous administration at a dose of 10 mg/kg/day for 8 days allowed survival of 20 % of infected mice, thus demonstrating the high potential of tetraoxanes for the treatment of Apicomplexan parasites.

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