Entamoeba histolytica: antigenic characterization of axenic strains from Brazil

Guimarães, Semíramis; Urdaneta, Haideé; Silva, Edward F.; Tavares, Carlos Alberto Pereira

doi:10.1590/s0036-46651991000100002

Cited by 4 publications

(2 citation statements)

References 16 publications

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“…E. histolytica trophozoites were axenically cultured in TYI-S-33 medium and harvested in the exponential growth phase (Diamond et al, 1978). E. histolytica clones A (virulent) and L-6 (non-virulent) were obtained from the HM1-IMSS strain (Orozco et al, 1983), whereas the clone 462 (virulent) came from a Brazilian patient (Guimaraes et al, 1991). Clone A was used for all experiments except when indicated.…”

Section: Methodsmentioning

confidence: 99%

Ehp53, an Entamoeba histolytica protein, ancestor of the mammalian tumour suppressor p53

et al. 2003

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This paper reports the identification of Ehp53, a p53-like Entamoeba histolytica protein, which binds to the human p53 DNA consensus sequence (oli-p53). Monoclonal antibodies against p53 (Ab-1 and Ab-2) recognized a single 53 kDa spot in two-dimensional gels and inhibited the formation of complexes produced by E. histolytica nuclear extracts and oli-p53. Additionally, E. histolytica gene promoter sequences with high homology to oli-p53 formed complexes with nuclear proteins that were abolished by oli-p53. Ehp53 protein levels increased in UV-irradiated trophozoites. This protein was also detected in Entamoeba moshkovskii and Entamoeba invadens. By confocal microscopy, Ehp53 was located in the nuclei, EhkO organelles and cytoplasm. The Ehp53-encoding gene was cloned and its predicted amino acid sequence showed 30-54 % and 50-57 % homology with important domains of the human and the Drosophila melanogaster p53 proteins, respectively. This homology included the tetramerization domain, the nuclear export signal and a nuclear localization signal. Ehp53 also contains seven of the eight DNA-binding residues and two of the four Zn 2+ -binding sites described for p53. A recombinant Ehp53 was recognized by Ab-2. Ehp53 is believed to be the first p53-like protein found in protozoa and may be the evolutionary ancestor of the mammalian p53.

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Section: Methodsmentioning

confidence: 99%

Ehp53, an Entamoeba histolytica protein, ancestor of the mammalian tumour suppressor p53

et al. 2003

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“…Preparation of anti-Giardia IgG -Immune sera to Giardia were raised by immunizing rabbits according to GUIMARÃES et al 8 . For each strain, two rabbits were immunized with four doses and the interval between each one was fifteen days.…”

Section: Methodsmentioning

confidence: 99%

Analysis of proteins from membrane and soluble fractions of Giardia duodenalis trophozoites of two Brazilian axenic strains

Guimarães

Sogayar

Franco

2002

Rev. Inst. Med. trop. S. Paulo

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SUMMARYIn the present study, we have analyzed by sodium docecyl sulphate -polyacrilamide gel electrophoresis (SDS-PAGE), immunoblotting and Concanavalin A blotting (Con A blotting) proteins of membrane fractions and soluble fractions obtained from Giardia duodenalis trophozoites of two axenic strains isolated in Brazil from a symptomatic (BTU-11) and an asymptomatic patient (BTU-10), as compared to the reference strain Portland 1. Both Brazilian strains showed a complex and homogeneous electrophoretic pattern of proteins, but some differences could be observed. Several glycoproteins were detected, particularly the proteins of 81, 72, 59 kDa and the protein of 62 kDa in the membrane proteins and cytosol, respectively. Many antigenic components were revealed by anti-Giardia rabbit IgG antibodies in the immunoblotting analysis. Among these components, the membrane protein of 32 kDa and the cytosol protein of 30 kDa could be related to giardin, as previously demonstrated.

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Entamoeba histolytica: detection of coproantigens by purified antibody in the capture sandwich ELISA

Urdaneta

Guimarães

Silva³

et al. 1994

Rev. Inst. Med. trop. S. Paulo

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A sensitive and specific Capture Sandwich ELISA (CSE) was developed using polyclonal purified rabbit antibodies against three different axenic strains of Entamoeba histolytica: CSP from Brazil and HM1 - IMSS from Mexico, for the detection of coproantigens in fecal samples. Immunoglobulin G (IgG) against E. histolytica was isolated from rabbits immunized with throphozoites whole extract in two stages: affinity chromatography in a column containing E. histolytica antigens bound to Sepharose 4B was followed by another chromatography in Sepharose antibodies 4B-Protein A. A Capture Sandwich ELISA using purified antibodies was able to detect 70ng of amebae protein, showing a sensitivity of 93% and specificity of 94%. The combination of microscopic examination and CSE gave a concordance and discordance of 93.25% and 6.75%, respectively. It was concluded that CSE is highly specific for the detection of coproantigens of E. histolytica in feces of infected patients, is quicker to perform, easier and more sensitive than microscopic examination.

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Entamoeba histolytica: antigenic characterization of axenic strains from Brazil

Cited by 4 publications

References 16 publications

Ehp53, an Entamoeba histolytica protein, ancestor of the mammalian tumour suppressor p53

Ehp53, an Entamoeba histolytica protein, ancestor of the mammalian tumour suppressor p53

Analysis of proteins from membrane and soluble fractions of Giardia duodenalis trophozoites of two Brazilian axenic strains

Entamoeba histolytica: detection of coproantigens by purified antibody in the capture sandwich ELISA

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