Dissecting the membrane cholesterol requirement for mycobacterial entry into host cells

Viswanathan, Gopinath; Jafurulla, Md.; Kumar, G. Aditya; Raghunand, Tirumalai R.; Chattopadhyay, Amitabha

doi:10.1016/j.chemphyslip.2015.05.006

Cited by 24 publications

(25 citation statements)

References 66 publications

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“…In this overall scenario, we previously demonstrated the requirement of membrane cholesterol in infection by Leishmania donovani [16,17,21] and Mycobacterium smegmatis [37]. These results are supported by work from other groups [23,46].…”

Section: Discussionsupporting

confidence: 82%

“…6). We have recently shown that such dependence on optimal host membrane cholesterol could be a hallmark of mycobacterial entry into host cells [37]. Whether this holds good for other intracellular pathogens whose entry is regulated by host membrane cholesterol requires careful examination.…”

Section: Discussionmentioning

confidence: 99%

“…Macrophage membrane cholesterol was enriched over its basal levels using the water-soluble MβCD-cholesterol complex as described previously [37] with some modifications. Briefly, the required amount of MβCD and cholesterol in the ratio of 10:1 (5 mM:0.5 mM) were dissolved in IMDM medium without serum under constant vortexing at room temperature (~25°C).…”

Section: Cholesterol Enrichment Using Mβcd-cholesterol Complexmentioning

confidence: 99%

“…Viability of macrophages upon treatment with lovastatin or MβCD-cholesterol complex was assessed using MTT assay as described earlier [24,37].…”

Section: Mtt Viability Assaymentioning

confidence: 99%

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Statin-induced chronic cholesterol depletion inhibits Leishmania donovani infection: Relevance of optimum host membrane cholesterol

Kumar

Roy

Jafurulla

et al. 2016

Biochimica et Biophysica Acta (BBA) - Biomembranes

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Leishmania are obligate intracellular protozoan parasites that invade and survive within host macrophages leading to leishmaniasis, a major cause of mortality and morbidity worldwide, particularly among economically weaker sections in tropical and subtropical regions. Visceral leishmaniasis is a potent disease caused by Leishmania donovani. The detailed mechanism of internalization of Leishmania is poorly understood. A basic step in the entry of Leishmania involves interaction of the parasite with the host plasma membrane. In this work, we have explored the effect of chronic metabolic cholesterol depletion using lovastatin on the entry and survival of Leishmania donovani in host macrophages. We show here that chronic cholesterol depletion of host macrophages results in reduction in the attachment of Leishmania promastigotes, along with a concomitant reduction in the intracellular amastigote load. These results assume further relevance since chronic cholesterol depletion is believed to mimic physiological cholesterol modulation. Interestingly, the reduction in the ability of Leishmania to enter host macrophages could be reversed upon metabolic replenishment of cholesterol. Importantly, enrichment of host membrane cholesterol resulted in reduction in the entry and survival of Leishmania in host macrophages. As a control, the binding of Escherichia coli to host macrophages remained invariant under these conditions, thereby implying specificity of cholesterol requirement for effective leishmanial infection. To the best of our knowledge, these results constitute the first comprehensive demonstration that an optimum content of host membrane cholesterol is necessary for leishmanial infection. Our results assume relevance in the context of developing novel therapeutic strategies targeting cholesterol-mediated leishmanial infection.

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Section: Discussionsupporting

confidence: 82%

Section: Discussionmentioning

confidence: 99%

Section: Cholesterol Enrichment Using Mβcd-cholesterol Complexmentioning

confidence: 99%

“…Viability of macrophages upon treatment with lovastatin or MβCD-cholesterol complex was assessed using MTT assay as described earlier [24,37].…”

Section: Mtt Viability Assaymentioning

confidence: 99%

See 2 more Smart Citations

Statin-induced chronic cholesterol depletion inhibits Leishmania donovani infection: Relevance of optimum host membrane cholesterol

Kumar

Roy

Jafurulla

et al. 2016

Biochimica et Biophysica Acta (BBA) - Biomembranes

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“…An effective way to achieve this is to reduce (or block) host membrane cholesterol. This approach has been successfully tried for entry of pathogens such as Leishmania [9,20,21,36,45,47,48,55] and Mycobacterium [12,20,57]. An advantage of such therapeutic approach involving modulation of host membrane cholesterol provides The energy landscape corresponding to the interaction of cholesterol with specific sites on the receptor surface involves a series of shallow minima connected by low energy barriers of the order of kT at room temperature.…”

Section: Role Of Host Membrane Cholesterol In Pathogen Entrymentioning

confidence: 99%

Cholesterol: An evergreen molecule in biology

Kumar

Chattopadhyay

2016

BSI

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Abstract. Cholesterol, an essential component of higher eukaryotic membranes, was discovered more than two centuries ago. The development and progress of cholesterol research in the last 200 years has been truly fascinating, with elements of surprise, serendipity and intrigue. In this review, we trace this journey the way we see it, and follow it up with the role of membrane cholesterol in crucial areas of contemporary research (transbilayer domains, regulation of GPCR function and role in the entry of intracellular pathogens into host cells), with considerable footprint from our work. We believe that cholesterol will continue to surprise and fascinate future researchers, thereby justifying its evergreen nature. Keywords: Membrane cholesterol, transbilayer dimer, G protein-coupled receptor, pathogen entry Abbreviations 7-DHC7-dehydrocholesterol 7-DHCR 3β-hydroxy-steroid-7 -reductase 25-NBD-cholesterol 25-[N-[(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-methyl]amino]-27-norcholesterol CRAC cholesterol recognition/interaction amino acid consensus DHE dehydroergosterol ( 5,7,9(11)22 -ergostatetraen-3β-ol) DPPC 1,2-dipalmitoyl-sn-glycero-3-phosphocholine FRAP fluorescence recovery after photobleaching GPCR G protein-coupled receptor NBD 7-nitrobenz-2-oxa-1,3-diazol-4-yl REES red edge excitation shift SLOS Smith-Lemli-Opitz syndrome

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Quantifying intracellular Mycobacterium tuberculosis: An essential issue for in vitro assays

et al. 2018

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Many studies about intracellular microorganisms which are important regarding diseases affecting public health have been focused on the recognition of host–pathogen interactions, thereby ascertaining the mechanisms by which the pathogen invades a cell and makes it become its host. Such knowledge enables understanding the immunological response triggered by these interactions for obtaining useful information for developing vaccines and drugs. Quantitative cell infection assay protocols are indispensable regarding studies involving Mycobacterium tuberculosis, which takes the lives of more than 2 million people worldwide every year; however, sometimes these are limited by the pathogen's slow growth. Concerning such limitation, a detailed review is presented here regarding the different methods for quantifying and differentiating an intracellular pathogen, the importance of mycobacteria aggregate dissociation and multiplicity of infection (MOI) in infection assays. The methods’ differences, advantages, and disadvantages are discussed regarding intra and extracellular bacteria (on cell surface) differentiation, current problems are outlined, as are the solutions provided using fluorophores and projections made concerning quantitative infection assays.

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Dissecting the membrane cholesterol requirement for mycobacterial entry into host cells

Cited by 24 publications

References 66 publications

Statin-induced chronic cholesterol depletion inhibits Leishmania donovani infection: Relevance of optimum host membrane cholesterol

Statin-induced chronic cholesterol depletion inhibits Leishmania donovani infection: Relevance of optimum host membrane cholesterol

Cholesterol: An evergreen molecule in biology

Quantifying intracellular Mycobacterium tuberculosis: An essential issue for in vitro assays

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