Isolation of Trypanosoma cruzi from Blood

Budzko, Delia B.; Kierszenbaum, Felipe

doi:10.2307/3278544

Cited by 61 publications

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“…The trypomastigotes were purified by centrifugation over Isolymph (Gallard Schlesinger, Carle Place, NY) (27) followed by chromatography through a DEAE-cellulose column (28). The eluted parasites were concentrated by centrifugation (800 x g, 15 min, 200C) and resuspended at 3-5 x 106 organisms per ml in the desired medium.…”

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confidence: 99%

Arginine decarboxylase inhibitors reduce the capacity of Trypanosoma cruzi to infect and multiply in mammalian host cells.

Kierszenbaum

Wirth

McCann

et al. 1987

Proc. Natl. Acad. Sci. U.S.A.

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Arginine decarboxylase inhibitors reduce the capacity of Trypanosoma cruzi to infect and multiply in mammalian host cells.

Kierszenbaum

Wirth

McCann

et al. 1987

Proc. Natl. Acad. Sci. U.S.A.

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“…Trypomastigotes were purified 10 to 12 days later from the blood by centrifugation through Isolymph (Gallard-Schlesinger, Carle Place, N.Y.) (5) followed by column chromatography through DEAEcellulose (15). The eluted trypanosomes were washed twice with Dulbecco's modified minimal essential medium (GIBCO, Grand Island, N.Y.) containing 100 IU of penicillin and 100 tg of streptomycin per ml and supplemented with 10% fetal bovine serum (Sigma Chemical Co., St. Louis, * Corresponding author.…”

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confidence: 99%

Inhibition of host cell invasion and intracellular replication of Trypanosoma cruzi by N,N'-bis(benzyl)-substituted polyamine analogs

Majumder

Kierszenbaum

1993

Antimicrob Agents Chemother

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We studied the effects of two N,N'-bis(benzyl)-substituted polyamine analogs on the capacities of Trypanosoma cruzi to invade and multiply within a mammalian host cell. At concentrations as low as 1 ,uM, these compounds reduced significantly the infectivity of the parasite for rat heart myoblasts in a time-dependent manner. Pretreatment of virulent T. cruzi trypomastigotes, but not myoblast pretreatment, reduced the level of infectivity. The inhibitory effects started to subside 3 h after removal of the drugs and were no longer detectable after 4 h. A significant decrease in the rate of intracellular amastigote multiplication was also seen when the drugs were added to myoblast cultures which had been previously infected with untreated T. cruzi.These results show that N,N'-bis(benzyl)-substituted polyamine analogs meet the two most important criteria for potential chemotherapeutic agents against T. cruzi infection, namely, inhibition of both host cell invasion and intracellular replication by this parasite.The unicellular parasite Trypanosoma cnrzi, the causative agent of Chagas' disease, requires an intracellular localization to multiply in mammalian hosts. This stresses the importance of developing drugs that can inhibit host cell invasion and subsequent intracellular parasite replication. Such drugs are badly needed given the lack of a consistently effective, nontoxic drug to cure Chagas' disease (9). There is evidence that both host cell invasion by T. cruzi trypomastigotes and intracellular multiplication of the amastigote form are impaired when production of the polyamines agmatine and putrescine is decreased by the specific arginine decarboxylase inhibitor DL-a-dlfluoromethylarginine (7,13).Polyamines play key roles in cell growth regulation, and polyamine synthesis inhibitors have been shown to affect some parasitic organisms (1,8,16). N,N'-Di-substituted tetraamines with the general formula RNH(CH2)XNH (CH2)YNH(CH2)NINHR have been reported to suppress the growth of several pathogenic protozoans (e.g., Leishmania donovani, Plasmodium falciparum, and Plasmodium berghei) in vitro and in infected animals (2-4) and could be suitable for exploring the role that polyamines and polyamine biosynthesis plays in T. cnuzi infection. In this work, we used two N,N'-bis(benzyl)-substituted polyamine analogs and studied their effects on T. cnuzi infectivity and replication using an in vitro model system of mammalian host cell infection.Stock CD1(ICR) Swiss mice (Charles River Laboratory, Portage, Mich.) were infected subcutaneously with 106 T. cruzi (Tulahen isolate) organisms. Trypomastigotes were purified 10 to 12 days later from the blood by centrifugation through Isolymph (Gallard-Schlesinger, Carle Place, N.Y.) (5) followed by column chromatography through DEAEcellulose (15). The eluted trypanosomes were washed twice with Dulbecco's modified minimal essential medium (GIBCO, Grand Island, N.Y.) containing 100 IU of penicillin and 100 tg of streptomycin per ml and supplemented with 10% fetal bovine serum (Sigma Chemi...

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“…For example, we have shown enhanced mitochondrial production of ROS in cardiomyocytes during T. cruzi infection (33). It is plausible that T. cruzi utilizes antioxidant enzyme network to prevent mitochondrial ROS-mediated direct killing or ROS activation of the apoptosis pathway for removal of the infected (or damaged) cells (57), thereby ensuring its persistence in the host. A consequence of prolonged survival of infected (damaged) cells is the signaling of cell repair and proliferative responses, culminating in cardiac fibrosis (58).…”

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TcI Isolates of Trypanosoma cruzi Exploit the Antioxidant Network for Enhanced Intracellular Survival in Macrophages and Virulence in Mice

et al. 2016

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h Trypanosoma cruzi species is categorized into six discrete typing units (TcI to TcVI) of which TcI is most abundantly noted in the sylvatic transmission cycle and considered the major cause of human disease. In our study, the TcI strains Colombiana (COL), SylvioX10/4 (SYL), and a cultured clone (TCC) exhibited different biological behavior in a murine model, ranging from high parasitemia and symptomatic cardiomyopathy (SYL), mild parasitemia and high tissue tropism (COL), to no pathogenicity (TCC). Proteomic profiling of the insect (epimastigote) and infective (trypomastigote) forms by two-dimensional gel electrophoresis/matrix-assisted laser desorption ionization-time of flight mass spectrometry, followed by functional annotation of the differential proteome data sets (>2-fold change, P < 0.05), showed that several proteins involved in (i) cytoskeletal assembly and remodeling, essential for flagellar wave frequency and amplitude and forward motility of the parasite, and (ii) the parasite-specific antioxidant network were enhanced in COL and SYL (versus TCC) trypomastigotes. Western blotting confirmed the enhanced protein levels of cytosolic and mitochondrial tryparedoxin peroxidases and their substrate (tryparedoxin) and iron superoxide dismutase in COL and SYL (versus TCC) trypomastigotes. Further, COL and SYL (but not TCC) were resistant to exogenous treatment with stable oxidants (H 2 O 2 and peroxynitrite [ONOO ؊ ]) and dampened the intracellular superoxide and nitric oxide response in macrophages, and thus these isolates escaped from macrophages. Our findings suggest that protein expression conducive to increase in motility and control of macrophage-derived free radicals provides survival and persistence benefits to TcI isolates of T. cruzi. C hagas disease, caused by the unicellular protozoan Trypanosoma cruzi, is ranked as the third most important parasitic disease in terms of disability-adjusted life years (1, 2). T. cruzi is naturally transmitted by Triatomine insects. In recent years, a significant proportion of the infected population has emigrated from rural areas, leading to the urbanization of Chagas disease in countries where the disease is endemic, as well as internationally (1). Chagas disease is therefore an emergent global public health problem associated with congenital transmission (3, 4), blood transfusions (5), and organ transplantations (6). Current estimates suggest that ϳ70 million people are at risk of infection (7,8) and that ϳ6 million individuals are infected with T. cruzi in Latin America and Mexico. However, these estimates may not be accurate, since they are not derived from detailed epidemiological studies. For example, in Argentina, the Chagas Disease National Control Program reported 13 provinces located in the Central-Northeast area as endemic for Chagas disease in 2010 (9). Vectorial transmission was reported to be absent in Jujuy, Entre Ríos, La Pampa, Neuquén, and Río Negro but active in provinces such as Formosa, Chaco, Córdoba, Santiago del Estero, and others, with a ...

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Isolation of Trypanosoma cruzi from Blood

Cited by 61 publications

References 0 publications

Arginine decarboxylase inhibitors reduce the capacity of Trypanosoma cruzi to infect and multiply in mammalian host cells.

Arginine decarboxylase inhibitors reduce the capacity of Trypanosoma cruzi to infect and multiply in mammalian host cells.

Inhibition of host cell invasion and intracellular replication of Trypanosoma cruzi by N,N'-bis(benzyl)-substituted polyamine analogs

TcI Isolates of Trypanosoma cruzi Exploit the Antioxidant Network for Enhanced Intracellular Survival in Macrophages and Virulence in Mice

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