Trypanosoma cruzi: Isolation and characterization of membrane and flagellar fractions

Pereira, Neize M.; Timm, S.L.; Costa, Salvatore; Rebello, M. A.; Souza, Wanderley de

doi:10.1016/0014-4894(78)90135-2

Cited by 24 publications

(24 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The material was centrifuged at 100,000g for 30 minutes in a swing out rotor on a Beckman ultracentrifuge, and the microsomal fraction (MicF) was recovered from the pellet. All fractions were suspended in PBS pH 7.2 and stored at −70°C (Pereira et al, 1978). Protein yield was determined by Bradford's method (Bradford, 1976).…”

Section: Antigenmentioning

confidence: 99%

“…Nevertheless, Pereira et al (1978) described a method to obtain highly pure fractions from Trypanosoma cruzi parasites.…”

Section: Introductionmentioning

confidence: 99%

“…In this article we have modified the fractionation method proposed by Pereira et al (1978) to obtain 3 L. (L.) amazonensis fractions that were characterized by transmission electron microscopy, SDS-gel electophoresis profile and immunogenicity.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

ExperimentalLeishmania (L.) amazonensisLeishmaniasis: Characterization and Immunogenicity of Subcellular Fractions

Valle

Gaspar

Souza-Lemos

et al. 2007

Immunological Investigations

View full text Add to dashboard Cite

A technique developed in Trypanosoma cruzi biochemical studies was successfully used to fractionate Leishmania (Leishmania) amazonensis promastigotes. Ultrastructural analyses revealed a membrane fraction (MF) associated to subpellicular microtubules, a ribosomal-rich microsomal fraction (MicF), and a flagellar fraction (FF) free of associated membrane. All fractions proved to be immunogenic through delayed type hypersensitivity reaction assays. Therefore, a protocol was designed to test whether these fractions could elicit a protective response in mice infected by L. (L), amazonensis. The protocol consisted of a BCG injection (as cellular immunity inducer), followed by cyclophosphamide (once its cytotoxic effect is over, this immunosuppressor can increase the number of circulating leukocytes), then an injection with one of the fractions followed by a challenge. When compared to infected control animals, mice injected with any of the fractions presented a smaller footpad swelling, especially those injected with MicF or FF. Macroscopically, immunized mice under modulation by BCG presented no swelling. Histopathological studies performed on day 120 revealed fewer amastigotes and more intense inflammation in lesions of MicF and FF injected mice. Animals injected with MF presented an intermediate pattern. Parasite quantification corroborated these results. The results show that all fractions are potent immunostimulators, but MicF and FF have the strongest protective ability.

show abstract

Section: Antigenmentioning

confidence: 99%

“…Nevertheless, Pereira et al (1978) described a method to obtain highly pure fractions from Trypanosoma cruzi parasites.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

ExperimentalLeishmania (L.) amazonensisLeishmaniasis: Characterization and Immunogenicity of Subcellular Fractions

Valle

Gaspar

Souza-Lemos

et al. 2007

Immunological Investigations

View full text Add to dashboard Cite

show abstract

“…This association of isolated membranes with microtubules has been confirmed for L. collosoma (Linder & Staehelin 1977), Trypanosoma cruzi (Meyer & De Souza 1976), Trypanosoma brucei (Voorheis et al 1979), Leishmania mexicana (Timm et al 1980), Leishmania donovani (Dwyer 1980), and Trypanosoma lewisi (Dywer & D'Alessandro 1980). In some cases, the microtubules did not remain associated with the plasma membrane (Pereira et al 1978, Franco da Silveira et al 1979, Zingales et al 1979, Rovis & Baekkeskov 1980. In these cases the only way to assess the purity of the fraction is to use the membrane enzyme markers or the attachment of microtubules to the isolated membranes.…”

Section: Isolation Of the Plasma Membranementioning

confidence: 99%

“…Since different methods have been used to rupture the cells, we cannot exclude the possibility that, in some cases, the attachment of microtubules to the membrane depends on differences in the methods used. But even when the same method was used for the isolation of the membrane, it gave different results in T. cruzi (Pereira et al 1978) and in L. mexicana (Timm et al 1980). Three procedures have been used to obtain a purified plasma membrane fraction from epimastigotes of T. cruzi.…”

Section: Isolation Of the Plasma Membranementioning

confidence: 99%

Cell fractionation of parasitic protozoa: a review

Souza

Cunha-e-Silva

2003

Mem. Inst. Oswaldo Cruz

View full text Add to dashboard Cite

Cell fractionation, a methodological strategy for obtaining purified organelle preparations, has been applied successfully to parasitic protozoa by a number of investigators. Here we present and discuss the work of several groups that have obtained highly purified subcellular fractions from trypanosomatids, Apicomplexa and trichomonads, and whose work have added substantially to our knowledge of the cell biology of these parasites.

show abstract

Flagellar Surfaces of Parasitic Protozoa and Their Role in Attachment

Vickerman

Tetley

1990

Ciliary and Flagellar Membranes

View full text Add to dashboard Cite

Trypanosoma cruzi: Isolation and characterization of membrane and flagellar fractions

Cited by 24 publications

References 28 publications

ExperimentalLeishmania (L.) amazonensisLeishmaniasis: Characterization and Immunogenicity of Subcellular Fractions

ExperimentalLeishmania (L.) amazonensisLeishmaniasis: Characterization and Immunogenicity of Subcellular Fractions

Cell fractionation of parasitic protozoa: a review

Flagellar Surfaces of Parasitic Protozoa and Their Role in Attachment

Contact Info

Product

Resources

About