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DOI | 10.1016/j.exppara.2019.01.012 |
Techniques to study phagocytosis and uptake of Leishmania tarentolae by J774 macrophages | |
Geroldinger, Gerald1; Rezk, Marlene1; Idris, Rugaia1; Gruber, Victoria1; Tonner, Matthias1; Moldzio, Rudolf2; Staniek, Katrin1; Monzote, Lianet3; Gille, Lars1 | |
发表日期 | 2019 |
ISSN | 0014-4894 |
EISSN | 1090-2449 |
卷号 | 197页码:57-64 |
英文摘要 | Leishmaniasis is a vector borne parasitic disease affecting millions of people worldwide and is spreading into further areas because of global warming. The development of new active substances against these single-cell eukaryotic parasites is of great importance. Leishmania tarentolae promastigotes (LtP) are non-pathogenic for mammals and serve as model organisms for pathogenic Leishmania in basic research. However, it is important to refine methods to study the process of the infection of mammalian macrophages by LtP and pathogenic Leishmania. Important stages of the infection are phagocytosis by macrophages and multiplication of Leishmania amastigotes in the phagolysosome of macrophages. In this study, advanced methods using electron spin resonance (ESR) spectroscopy and genetically manipulated LtP were used to monitor the infection of adherent J774 macrophages with LtP. An ESR method was established to detect the formation of superoxide radicals directly in adherent J774 cells and to investigate the effect of LtP on this activity. J774 cells responded with a burst of superoxide radicals in the presence of phorbol myristate acetate as positive control. In contrast, challenging J774 cells with LtP resulted in a much lower burst of superoxide radicals. To facilitate LtP detection in the phagolysosome of J774 macrophages, LtP expressing enhanced green fluorescent protein (EGFP-LtP) were constructed. After different infection times with EGFP-LtP, the J774 cells were visualized by phase contrast microscopy and the cell number was determined. The intramacrophage Leishmania tarentolae amastigotes (LtA) expressing EGFP were detected by fluorescence microscopy and then counted with ImageJ. These experiments showed that LtP are taken up by J774 cells and form intraphagolysosomal amastigotes. LtA under our conditions multiplied intracellularly and were able to persist about 48 h in J774 cells. These experiments showed that ESR spectroscopy of attached macrophages and the use of the EGFP-LIP are suitable methods to study the initial phase of Leishmania infection in vitro. |
WOS研究方向 | Parasitology |
来源期刊 | EXPERIMENTAL PARASITOLOGY |
文献类型 | 期刊论文 |
条目标识符 | http://gcip.llas.ac.cn/handle/2XKMVOVA/92623 |
作者单位 | 1.Univ Vet Med, Dept Biomed Sci, Inst Pharmacol & Toxicol, Vienna, Austria; 2.Univ Vet Med, Dept Biomed Sci, Inst Med Biochem, Vienna, Austria; 3.Inst Trop Med Pedro Kouri, Dept Parasitol, Havana, Cuba |
推荐引用方式 GB/T 7714 | Geroldinger, Gerald,Rezk, Marlene,Idris, Rugaia,et al. Techniques to study phagocytosis and uptake of Leishmania tarentolae by J774 macrophages[J],2019,197:57-64. |
APA | Geroldinger, Gerald.,Rezk, Marlene.,Idris, Rugaia.,Gruber, Victoria.,Tonner, Matthias.,...&Gille, Lars.(2019).Techniques to study phagocytosis and uptake of Leishmania tarentolae by J774 macrophages.EXPERIMENTAL PARASITOLOGY,197,57-64. |
MLA | Geroldinger, Gerald,et al."Techniques to study phagocytosis and uptake of Leishmania tarentolae by J774 macrophages".EXPERIMENTAL PARASITOLOGY 197(2019):57-64. |
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