An Isoniazid Analogue Promotes Mycobacterium tuberculosis-Nanoparticle Interactions and Enhances Bacterial Killing by Macrophages

Faria, Tatiany J. de; Roman, Mariane; Souza, Nicole Menezes de; Vecchi, Rodrigo De; Assis, João Vitor de; Santos, Ana Lúcia Gomes dos; Bechtold, Ivan H.; Winter, Nathalie; Soares, Maurílio J.; Silva, Luciano; Almeida, Mauro Vieira de; Báfica, André

doi:10.1128/aac.05993-11

Cited by 51 publications

(40 citation statements)

References 48 publications

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“…Our data show that when either mouse primary macrophages or RAW cells were infected with red BCG and allowed to internalize the coumarin-6 green-labeled particles, there was no evident colocalization between the bacteria and the PLGA particles. This result is in agreement with the conclusions made by (Kisich et al, 2007), but at odds with the findings of (de Faria et al, 2012) who concluded that PLGA NPs enclosing an isoniazid derivative directly interacts with the BCG phagosomes in macrophages. Earlier studies had established that BCG, like M.tb, resides in an arrested early phagosome stage, with only a small fraction of phagosomes acquiring markers of lysosomes.…”

Section: Discussioncontrasting

confidence: 50%

Polylactide-co-glycolide-rifampicin-nanoparticles efficiently clearMycobacterium bovisBCG infection in macrophages and remain membrane-bound in phago-lysosomes

Kalluru

Fenaroli

Westmoreland

et al. 2013

Journal of Cell Science

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Summary Nanoparticles (NPs) are increasingly used as biodegradable vehicles to selectively deliver therapeutic agents such as drugs or antigens to cells. The most widely used vehicle for this purpose is based on copolymers of lactic acid and glycolic acid (PLGA) and has been extensively used in experiments aimed at delivering antibiotics against Mycobacterium tuberculosis in animal models of tuberculosis. Here, we describe fabrication of PLGA NPs containing either a high concentration of rifampicin or detectable levels of the green fluorescent dye, coumarin-6. Our goal here was twofold: first to resolve the controversial issue of whether, after phagocytic uptake, PLGA NPs remain membrane-bound or whether they escape into the cytoplasm, as has been widely claimed. Second, we sought to make NPs that enclosed sufficient rifampicin to efficiently clear macrophages of infection with Mycobacterium bovis BCG. Using fluorescence microscopy and immuno-electron microscopy, in combination with markers for lysosomes, we show that BCG bacteria, as expected, localized to early phagosomes, but that at least 90% of PLGA particles were targeted to, and remained in, low pH, hydrolaserich phago-lysosomes. Our data collectively argue that PLGA NPs remain membrane-enclosed in macrophages for at least 13 days and degrade slowly. Importantly, provided that the NPs are fabricated with sufficient antibiotic, one dose given after infection is sufficient to efficiently clear the BCG infection after 9-12 days of treatment, as shown by estimates of the number of bacterial colonies in vitro.

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Section: Discussioncontrasting

confidence: 50%

Polylactide-co-glycolide-rifampicin-nanoparticles efficiently clearMycobacterium bovisBCG infection in macrophages and remain membrane-bound in phago-lysosomes

Kalluru

Fenaroli

Westmoreland

et al. 2013

Journal of Cell Science

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“…The active INH content of the NPs could in future be adjusted to an effective concentration that takes into consideration both the slow drug releasing nature of the PLGA particle and the degree of targetting achievable by the MA ligandation. The concentrations of INH in the particles used for the EM observations were approximately 5 μg/ml in the preparation medium, ten times less than a recent in vitro study of an INH analogue that was tested in PLGA particles [47]. As mentioned before, INH-PLGA NPs have shown sustained drug release over several days in vitro and in vivo [66] and allowed a reduction in treatment frequency in mice and guinea pig models [67][68][69].…”

Section: Discussionmentioning

confidence: 94%

“…Another issue is the ability for PLGA particles to reach the compartment in which mycobacteria reside. One group suggested this possibility [47], but others could not demonstrate it [10,45].…”

Section: Introductionmentioning

confidence: 96%

Mycolic acids, a promising mycobacterial ligand for targeting of nanoencapsulated drugs in tuberculosis

Lemmer

Kalombo

Pietersen

et al. 2015

Journal of Controlled Release

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The appearance of drug-resistant strains of Mycobacterium tuberculosis (Mtb) poses a great challenge to the development of novel treatment programmes to combat tuberculosis. Since innovative nanotechnologies might alleviate the limitations of current therapies, we have designed a new nanoformulation for use as an anti-TB drug delivery system. It consists of incorporating mycobacterial cell wall mycolic acids (MA) as targeting ligands into a drug-encapsulating Poly dl-lactic-co-glycolic acid polymer (PLGA), via a double emulsion solvent evaporation technique. Bone marrow-derived mouse macrophages, either uninfected or infected with different mycobacterial strains (Mycobacterium avium, Mycobacterium bovis BCG or Mtb), were exposed to encapsulated isoniazid-PLGA nanoparticles (NPs) using MA as a targeting ligand. The fate of the NPs was monitored by electron microscopy. Our study showed that i) the inclusion of MA in the nanoformulations resulted in their expression on the outer surface and a significant increase in phagocytic uptake of the NPs; ii) nanoparticle-containing phagosomes were rapidly processed into phagolysosomes, whether MA had been included or not; and iii) nanoparticle-containing phagolysosomes did not fuse with non-matured mycobacterium-containing phagosomes, but fusion events with mycobacterium-containing phagolysosomes were clearly observed.

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“…Bacterial concentration was determined by a number 1 McFarland scale equivalent to 3 × 10 8 bacteria/mL. The INH resistance phenotype of 255R was confirmed by experiments employing a previously described MTT protocol [66], which revealed that the minimal inhibitory concentration (MIC) of 255R is 30 times higher than the H37Rv MIC. Presence of the functionally impaired S315T katG mutation was detected by sequencing.…”

mentioning

confidence: 99%

Isoniazid‐induced control of Mycobacterium tuberculosis by primary human cells requires interleukin‐1 receptor and tumor necrosis factor

et al. 2016

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Proinflammatory cytokines are critical mediators that control Mycobacterium tuberculosis (Mtb) growth during active tuberculosis (ATB). To further inhibit bacterial proliferation in diseased individuals, drug inhibitors of cell wall synthesis such as isoniazid (INH) areemployed. However, whether INH presents an indirect effect on bacterial growth by regulating host cytokines during ATB is not well known. To examine this hypothesis, we used an in vitro human granuloma system generated with primary leukocytes from healthy donors adapted to model ATB. Intense Mtb proliferation in cell cultures was associated with monocyte/macrophage activation and secretion of IL-1β and TNF. Treatment with INH significantly reduced Mtb survival, but altered neither T-cell-mediated Mtb killing, nor production of IL-1β and TNF. However, blockade of both IL-1R1 and TNF signaling rescued INH-induced killing, suggesting synergistic roles of these cytokines in mediating control of Mtb proliferation. Additionally, mycobacterial killing by INH was highly dependent upon drug activation by the pathogen catalase-peroxidase KatG and involved a host PI3K-dependent pathway. Finally, experiments using coinfected (KatG-mutated and H37Rv strains) cells suggested that active INH does not directly enhance host-mediated killing of Mtb. Our results thus indicate that Mtb-stimulated host IL-1 and TNF have potential roles in TB chemotherapy.Keywords: Human r IL-1 r TNF r Isoniazid r Leukocytes r Mycobacterium tuberculosis Additional supporting information may be found in the online version of this article at the publisher's web-site IntroductionUpon Mycobacterium tuberculosis (Mtb) exposure, it is estimated that 5-10% of human subjects will develop active tuberculosis Correspondence: Prof. André Báfica e-mail: andre.bafica@ufsc.br (TB) [1], which is characterized by intense pathogen proliferation associated with cellular activation and cell death. Cellular structures known as necrotic granulomas may promote bacterial dissemination and enhance Mtb transmission [2,3]. On the other hand, counter balance is supported by host recognition of Mtb, which induces cytokines such as IL-1β and TNF [4,5], two potent inflammatory mediators that suppress Mtb growth [4,6]. It has C 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.eji-journal.eu Eur. J. Immunol. 2016. 46: 1936-1947 Immunity to infection 1937 been shown that these cytokines regulate intracellular bacterial growth in human macrophages, whether by recruitment of antimicrobial effector molecules [7] or induction of apoptotic cell death [8]. In conjunction with 9], IL-1β, and TNF can provide signals to help differentiation and antigen presentation by macrophages [6,10,11]. Moreover, the adverse outcome of latent TB reactivation observed in clinical trials with anti-TNF therapies, additionally confirmed the essential role of TNF for controlling Mtb growth in humans [12]. In addition to antibiotics, which are major therapeutic tools to promote further suppression of mycobacterial growth [13], host-targeted i...

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An Isoniazid Analogue Promotes Mycobacterium tuberculosis-Nanoparticle Interactions and Enhances Bacterial Killing by Macrophages

Cited by 51 publications

References 48 publications

Polylactide-co-glycolide-rifampicin-nanoparticles efficiently clearMycobacterium bovisBCG infection in macrophages and remain membrane-bound in phago-lysosomes

Polylactide-co-glycolide-rifampicin-nanoparticles efficiently clearMycobacterium bovisBCG infection in macrophages and remain membrane-bound in phago-lysosomes

Mycolic acids, a promising mycobacterial ligand for targeting of nanoencapsulated drugs in tuberculosis

Isoniazid‐induced control of Mycobacterium tuberculosis by primary human cells requires interleukin‐1 receptor and tumor necrosis factor

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