EASY-HIT: HIV Full-Replication Technology for Broad Discovery of Multiple Classes of HIV Inhibitors

Kremb, Stephan; Helfer, Markus; Heller, Werner; Hoffmann, Dieter; Wolff, Horst; Kleinschmidt, A.; Cepok, Sabine; Hemmer, Bernhard; Durner, Jörg; Brack‐Werner, Ruth

doi:10.1128/aac.00515-10

Cited by 40 publications

(83 citation statements)

References 59 publications

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“…One hundred MΦs/coverslip were scored and the amastigotes were enumerated [19]. The average of three untreated cultures was taken as 100% control against which the percentage inhibition of infected MΦs in treated cultures was calculated.…”

Section: Methodsmentioning

confidence: 99%

Imipramine Is an Orally Active Drug against Both Antimony Sensitive and Resistant Leishmania donovani Clinical Isolates in Experimental Infection

Mukherjee

Mukhopadhyay

et al. 2012

PLoS Negl Trop Dis

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BackgroundIn an endeavor to find an orally active and affordable antileishmanial drug, we tested the efficacy of a cationic amphiphilic drug, imipramine, commonly used for the treatment of depression in humans. The only available orally active antileishmanial drug is miltefosine with long half life and teratogenic potential limits patient compliance. Thus there is a genuine need for an orally active antileishmanial drug. Previously it was shown that imipramine, a tricyclic antidepressant alters the protonmotive force in promastigotes, but its in vivo efficacy was not reported.Methodology/Principal FindingsHere we show that the drug is highly active against antimony sensitive and resistant Leishmania donovani in both promastigotes and intracellular amastigotes and in LD infected hamster model. The drug was found to decrease the mitochondrial transmembrane potential of Leishmania donovani (LD) promastigotes and purified amastigotes after 8 h of treatment, whereas miltefosine effected only a marginal change even after 24 h. The drug restores defective antigen presenting ability of the parasitized macrophages. The status of the host protective factors TNF α, IFN γ and iNOS activity increased with the concomitant decrease in IL 10 and TGF β level in imipramine treated infected hamsters and evolution of matured sterile hepatic granuloma. The 10-day therapeutic window as a monotherapy, showing about 90% clearance of organ parasites in infected hamsters regardless of their SSG sensitivity.ConclusionsThis study showed that imipramine possibly qualifies for a new use of an old drug and can be used as an effective orally active drug for the treatment of Kala-azar.

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Section: Methodsmentioning

confidence: 99%

Imipramine Is an Orally Active Drug against Both Antimony Sensitive and Resistant Leishmania donovani Clinical Isolates in Experimental Infection

Mukherjee

Mukhopadhyay

et al. 2012

PLoS Negl Trop Dis

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“…Cell-based HIV detection relies on molecular imaging techniques, such as immunofluorescence, and electron microscopy, which both allow direct visualization of viral structures but require cell fixation. Live cell reporter systems include the implementation of genetic reporter elements that get activated upon HIV infection [1], [2], [3] as well as recombinant viruses, where tags or fluorescent proteins have been integrated to study replication dynamics in living cells. In particular, HIV assembly processes in living cells have been a major subject of investigation over the last years (recently reviewed [4], [5]).…”

Section: Introductionmentioning

confidence: 99%

Direct and Dynamic Detection of HIV-1 in Living Cells

et al. 2012

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In basic and applied HIV research, reliable detection of viral components is crucial to monitor progression of infection. While it is routine to detect structural viral proteins in vitro for diagnostic purposes, it previously remained impossible to directly and dynamically visualize HIV in living cells without genetic modification of the virus. Here, we describe a novel fluorescent biosensor to dynamically trace HIV-1 morphogenesis in living cells. We generated a camelid single domain antibody that specifically binds the HIV-1 capsid protein (CA) at subnanomolar affinity and fused it to fluorescent proteins. The resulting fluorescent chromobody specifically recognizes the CA-harbouring HIV-1 Gag precursor protein in living cells and is applicable in various advanced light microscopy systems. Confocal live cell microscopy and super-resolution microscopy allowed detection and dynamic tracing of individual virion assemblies at the plasma membrane. The analysis of subcellular binding kinetics showed cytoplasmic antigen recognition and incorporation into virion assembly sites. Finally, we demonstrate the use of this new reporter in automated image analysis, providing a robust tool for cell-based HIV research.

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“…The MTT solution was carefully removed and the cells were lysed by adding 100 μL of lysis solution (20% [wt/v] sodium dodecyl sulfate +0.6% [v/v] 37% HCl in dimethyl sulfoxide). 47 The MTT formazan crystals were dissolved by gently tapping the plate, and their optical density (OD) was measured by a microplate reader (xMark TM microplate-absorbance-spectrophotometer; Bio-Rad Laboratories Inc., Hercules, CA, USA) at a test wavelength of 570 nm and a reference wavelength of 630 nm. The cell viability was calculated as the percentage of optical density of the treated cells compared with the negative control (NC), ie, the untreated cells:…”

Section: Cytotoxicity Assaysmentioning

confidence: 99%

Non-chemotoxic induction of cancer cell death using magnetic nanowires

Contreras¹,

Sougrat²,

Zaher³

et al. 2015

IJN

101

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In this paper, we show that magnetic nanowires with weak magnetic fields and low frequencies can induce cell death via a mechanism that does not involve heat production. We incubated colon cancer cells with two concentrations (2.4 and 12 μg/mL) of nickel nanowires that were 35 nm in diameter and exposed the cells and nanowires to an alternating magnetic field (0.5 mT and 1 Hz or 1 kHz) for 10 or 30 minutes. This low-power field exerted a force on the magnetic nanowires, causing a mechanical disturbance to the cells. Transmission electron microscopy images showed that the nanostructures were internalized into the cells within 1 hour of incubation. Cell viability studies showed that the magnetic field and the nanowires separately had minor deleterious effects on the cells; however, when combined, the magnetic field and nanowires caused the cell viability values to drop by up to 39%, depending on the strength of the magnetic field and the concentration of the nanowires. Cell membrane leakage experiments indicated membrane leakage of 20%, suggesting that cell death mechanisms induced by the nanowires and magnetic field involve some cell membrane rupture. Results suggest that magnetic nanowires can kill cancer cells. The proposed process requires simple and low-cost equipment with exposure to only very weak magnetic fields for short time periods.

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EASY-HIT: HIV Full-Replication Technology for Broad Discovery of Multiple Classes of HIV Inhibitors

Cited by 40 publications

References 59 publications

Imipramine Is an Orally Active Drug against Both Antimony Sensitive and Resistant Leishmania donovani Clinical Isolates in Experimental Infection

Imipramine Is an Orally Active Drug against Both Antimony Sensitive and Resistant Leishmania donovani Clinical Isolates in Experimental Infection

Direct and Dynamic Detection of HIV-1 in Living Cells

Non-chemotoxic induction of cancer cell death using magnetic nanowires

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