Differential gene expression during Trypanosoma cruzi metacyclogenesis

Krieger, Marco Aurélio; Ávila, Andréa Rodrigues; Ogatta, Sueli Fumie Yamada; Plazanet-Menut, Claire; Goldenberg, Samuel

doi:10.1590/s0074-02761999000700021

Cited by 22 publications

(15 citation statements)

References 16 publications

(14 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…mRNA from epimastigotes was isolated by use of the Dynabeads mRNA Direct kit, according to the manufacturer's instructions (Dynal Biotech). mRNA was quantified by spectrometry (at 260 nm) and was then retrotranscribed into single-strand (ss) cDNA by using an avian myeloblastosis virus reverse transcriptase (Promega) and an oligo(dT 18 ) primer. Reverse transcription was carried out in a PTC-100 Thermocycler (MJ Research) for 1 h at 42°C.…”

Section: Methodsmentioning

confidence: 99%

“…Reverse transcription was carried out in a PTC-100 Thermocycler (MJ Research) for 1 h at 42°C. Double-strand (ds) cDNA was generated by using a primer specific for the conserved 39 nucleotides (nt) at the 3Ј end of the miniexon SL of all Trypanosoma mRNAs (18), which contain 23 nt of the miniexon sequence and the oligo(dT 18 ) primer. The optimal conditions for ss cDNA amplification were 16 M SLc (5Ј-GATACAGTTTCTGTACTATATT G-3Ј), 16 M oligo(dT 18 ), 166.66 M deoxynucleoside triphosphates, 1 U Taq polymerase (Boehringer Mannhein), 10 l 10ϫ Tampon, and 100 ng ss cDNA in a 100-l total volume.…”

Section: Methodsmentioning

confidence: 99%

“…Among the different techniques that have been used to study the differential expression of genes involved in cellular drug resistance, the differential display method is one of the most widely used (15,54). Currently, it is possible to compare genes that are differentially expressed during the life cycle of parasites by studying mRNA polymorphism through RNA differential display (28) or representation of differential expression, which consists of the selection of specific genes through PCR amplification of hybrid selected sequences (18).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Differential Gene Expression in Benznidazole-Resistant Trypanosoma cruzi Parasites

Villarreal

Nirdé

Hide

et al. 2005

Antimicrob Agents Chemother

View full text Add to dashboard Cite

We analyzed the differential gene expression among representative Trypanosoma cruzi stocks in relation to benznidazole exposures using a random differentially expressed sequences (RADES) technique. Studies were carried out with drug pressure both at the natural susceptibility level of the wild-type parasite (50% inhibitory concentration for the wild type) and at different resistance levels. The pattern of differential gene expression performed with resistant stocks was compared to the population structure of this parasite, established by random amplified polymorphic DNA analysis and multilocus enzyme electrophoresis. A RADES band polymorphism was observed, and over- or underexpression was linked to the resistance level of the stock. The analysis of RADES bands suggested that different products may be involved in benznidazole resistance mechanisms. No significant association was found between phylogenetic clustering and benznidazole susceptibility. Benznidazole resistance may involve several mechanisms, depending on the level of drug exposure

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Differential Gene Expression in Benznidazole-Resistant Trypanosoma cruzi Parasites

Villarreal

Nirdé

Hide

et al. 2005

Antimicrob Agents Chemother

View full text Add to dashboard Cite

show abstract

“…However, in the mammal, differentiation into intracellular amastigote as well as extracellular trypomastigote forms occurs as stages in the life cycle [32]. These differentiation events have also been correlated with changes in gene expression [32][33][34][35][36].…”

Section: Introductionmentioning

confidence: 99%

The Application of Gene Expression Microarray Technology to Kinetoplastid Research

Duncan

Salotra²,

Goyal³

et al. 2004

CMM

View full text Add to dashboard Cite

Protozoan parasites in the order Kinetoplastida cause severe disease primarily in tropical and subtropical areas. Vaccines to control these diseases have shown some promise, but none are in active clinical use. Drug treatments are available for all of the acute infections, but the emergence of resistance and an unresponsive chronic phase are current problems. Rapid advances in genomic technology open the possibility of discovering new genes that can contribute to vaccine initiatives or serve as targets for development of new drugs. The DNA microarray is a genomic technology, which is being applied to new gene discovery in kinetoplastid parasites. Both cDNA and genomic microarrays for Leishmania major have identified a number of new genes that are expressed in a stage-specific fashion and preliminary results from a L. donovani genomic microarray also demonstrated new gene discovery. A microarray of Trypanosoma brucei genomic fragments identified new genes whose expression differs between the insect borne stage and the human infectious stage of the parasite. The next few years, building on this foundational work, should witness the most exciting stage as microarrays are applied to questions such as the basis of drug resistance, post kala azar dermal leishmaniasis, the regulation of differentiation to infectious stages, linking coordinately regulated pathways of genes and development of genetically defined parasites that may have potential as live attenuated vaccines.

show abstract

“…In addition, epithelial cells produced NO after infection by BFT but not after infection by MT. Multiple studies have demonstrated that T. cruzi parasites express stage-specific proteins and other molecules, including some that are generated by stage-specific posttranscriptional and posttranslational modifications (1,5,6,10,13,18,21). It is possible that the differential chemokine and NO responses induced in epithelial cells by BFT and MT are due to the distinct glycoconjugates present on their surface membranes shown previously to differentially induce cytokines and NO production in macrophages (3,4,8,9).…”

mentioning

confidence: 99%

Differential Interleukin-8 and Nitric Oxide Production in Epithelial Cells Induced by Mucosally Invasive and NoninvasiveTrypanosoma cruziTrypomastigotes

2003

View full text Add to dashboard Cite

Trypanosoma cruzi metacyclic trypomastigotes (MT), but not blood form trypomastigotes (BFT), are highly mucosally infective. We investigated the abilities of MT and BFT to induce inflammation and/or intracellular killing activity within mucosal epithelia. BFT, but not MT, induced marked increases in interleukin-8, GRO-␣, MCP-1, and nitric oxide production in HeLa and AGS cells, despite similar infectivities. MT may avoid induction of inflammation as an important biological mechanism facilitating mucosal invasion.An estimated 16 to 18 million people in South and Central America are infected with Trypanosoma cruzi, the protozoan parasite causing Chagas' disease (7). Fifty thousand deaths per year result from the cardiac and gastrointestinal pathology due to chronic T. cruzi infection. T. cruzi is most commonly transmitted to humans through contact with the infected feces of the reduviid bug. T. cruzi metacyclic trypomastigotes (MT) in reduviid bug feces can initiate infection through cuts in the skin or through contact with mucosal surfaces, including the eyes and mouth. T. cruzi MT infect epithelial cells, develop, and replicate intracellularly as amastigotes and are released from lysed host cells as blood form trypomastigotes (BFT). BFT are then able to infect new epithelial cells and most other types of mammalian host cells and repeat the intracellular replication cycle. T. cruzi MT are able to initiate infection through the conjunctival and gastrointestinal mucosa. On the other hand, we have shown that BFT are unable to infect through mucosal surfaces (14). The mechanisms responsible for the mucosal invasiveness of MT, but not BFT, are not known.Tissue infection with T. cruzi elicits significant inflammation responsible for the pathology associated with Chagas' disease. BFT, but not MT, infection of macrophages induces proinflammatory cytokine and nitric oxide (NO) production (8). The effects of T. cruzi infection on proinflammatory responses in epithelial cells, however, have not been reported. We found that mice challenged orally with MT do not develop detectable mucosal inflammation until 4 to 7 days after infection, sufficient time for three to four cycles of replication and 100-to 1,000-fold amplification of BFT (16). We hypothesize that the inability of MT to induce inflammatory mediators could be important for T. cruzi mucosal infectivity. On the other hand, it may be advantageous for later stages of infection with BFT to induce inflammation, prolonging the life of the host by preventing uncontrolled parasite replication.We studied the ability of T. cruzi strain Tulahuén MT and BFT to induce chemokine responses in mucosal epithelial cells. Culture-derived MT were prepared by subculture of axenically cultivated epimastigotes into modified Grace's medium (Sigma, St. Louis, Mo.) for 7 to 10 days. These culturederived MT are morphologically similar to insect-derived MT and are mucosally infective (20). Insect-derived MT were prepared by allowing T. cruzi-infected reduviid bugs (Dipetalogaster maximus) to fe...

show abstract

Differential gene expression during Trypanosoma cruzi metacyclogenesis

Cited by 22 publications

References 16 publications

Differential Gene Expression in Benznidazole-Resistant Trypanosoma cruzi Parasites

Differential Gene Expression in Benznidazole-Resistant Trypanosoma cruzi Parasites

The Application of Gene Expression Microarray Technology to Kinetoplastid Research

Differential Interleukin-8 and Nitric Oxide Production in Epithelial Cells Induced by Mucosally Invasive and NoninvasiveTrypanosoma cruziTrypomastigotes

Contact Info

Product

Resources

About