Molecular cloning and characterization of an Echinococcus granulosus cDNA encoding malate dehydrogenase

Rodrigues, Jaqueline J.S.; Ferreira, Henrique Bunselmeyer; Zaha, Arnaldo

doi:10.1016/0166-6851(93)90040-5

Cited by 15 publications

(7 citation statements)

References 8 publications

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“…A common panel constituted by the same batch of six E. granulosus antigens was evaluated by all participating laboratories and included bovine HCF (HCF 1 ); native AgB; two recombinant AgB subunits, namely, AgB8/1 (7) and AgB8/2 (6); recombinant cytosolic malate dehydrogenase from E. granulosus (18); and an AgB-derived synthetic peptide (p-176) (10). HCF was obtained by aseptic aspiration from either bovine or ovine fertile cyst (2).…”

Section: Methodsmentioning

confidence: 99%

Comparative Analysis of the Diagnostic Performance of Six Major Echinococcus granulosus Antigens Assessed in a Double-Blind, Randomized Multicenter Study

Lorenzo¹,

Ferreira

Monteiro

et al. 2005

J Clin Microbiol

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The serodiagnosis of hydatid disease is a valuable instrument for clinical diagnosis and epidemiological surveillance of high-risk populations. In the past decade a wealth of reports on the diagnostic performance of numerous antigens have been produced. However, their diagnostic value has been estimated under different conditions, using different serum collection, therefore precluding their direct comparison. Here we report an unbiased comparison of the same batch of six major E. granulosus antigens, namely, hydatid cyst fluid (HCF), native antigen B (AgB), two recombinant AgB subunits, an AgB-derived synthetic peptide, and recombinant cytosolic malate dehydrogenase from E. granulosus (EgMDH), against the same serum collection. The doubleblind analysis was performed using a standardized protocol and receiver operating characteristic (ROC) data analysis by a network of six South American laboratories. High intercenter reproducibility was attained, and the intralaboratory analysis allowed the comparative ranking of the antigen panel. HCF, AgB, and its AgB8/1 subunit exhibited equivalent diagnostic efficiencies, 81.4% ؎ 0.5%, 81.3% ؎ 0.6%, and 81.9% ؎ 2.0%, respectively; with a more favorable balance toward specificity in the case of the last antigen. The diagnostic efficiencies for the other three antigens were 76.8% ؎ 6.8%, 69.1% ؎ 2.7%, and 66.8% ؎ 2.1%, for the peptide, the AgB8/2 subunit, and the EgMDH, respectively. The study also included an analysis of batch-to-batch variation in the diagnostic performance of different HCF regional preparations. Based on these results, a suggested recommendation on the use of these antigens was drawn.

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Section: Methodsmentioning

confidence: 99%

Comparative Analysis of the Diagnostic Performance of Six Major Echinococcus granulosus Antigens Assessed in a Double-Blind, Randomized Multicenter Study

Lorenzo¹,

Ferreira

Monteiro

et al. 2005

J Clin Microbiol

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“…Both, in antigen characterization studies and in proteomic surveys, many cytosolic proteins, including glycolytic enzymes, were also consistently found in extracellular host-interacting components (e.g. tegument, and hydatid fluid) (Aziz et al, 2011;Chemale et al, 2003;Monteiro et al, 2010;Rodrigues et al, 1993), and the ectopic presence of these proteins was taken as evidence of potential moonlight functions.…”

Section: Introductionmentioning

confidence: 99%

Fructose-bisphosphate aldolase and enolase from Echinococcus granulosus: Genes, expression patterns and protein interactions of two potential moonlighting proteins

Lorenzatto

Monteiro

Paredes

et al. 2012

Gene

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Glycolytic enzymes, such as fructose-bisphosphate aldolase (FBA) and enolase, have been described as complex multifunctional proteins that may perform non-glycolytic moonlighting functions, but little is known about such functions, especially in parasites. We have carried out in silico genomic searches in order to identify FBA and enolase coding sequences in Echinococcus granulosus, the causative agent of cystic hydatid disease. Four FBA genes and 3 enolase genes were found, and their sequences and exon-intron structures were characterized and compared to those of their orthologs in Echinococcus multilocularis, the causative agent of alveolar hydatid disease. To gather evidence of possible non-glycolytic functions, the expression profile of FBA and enolase isoforms detected in the E. granulosus pathogenic larval form (hydatid cyst) (EgFBA1 and EgEno1) was assessed. Using specific antibodies, EgFBA1 and EgEno1 were detected in protoscolex and germinal layer cells, as expected, but they were also found in the hydatid fluid, which contains parasite's excretory-secretory (ES) products. Besides, both proteins were found in protoscolex tegument and in vitro ES products, further suggesting possible non-glycolytic functions in the host-parasite interface. EgFBA1 modeled 3D structure predicted a F-actin binding site, and the ability of EgFBA1 to bind actin was confirmed experimentally, which was taken as an additional evidence of FBA multifunctionality in E. granulosus. Overall, our results represent the first experimental evidences of alternative functions performed by glycolytic enzymes in E. granulosus and provide relevant information for the understanding of their roles in host-parasite interplay.

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“…Our laboratory, individually or in association with other research groups, has cloned several E. granulosus genes, among which are those coding for two different AgB subunits [18,19], a malate dehydrogenase [20,21], an actin filament fragmenting protein [22] and a calcium binding protein [23]. These proteins were expressed in Escherichia coli and the recombinant antigens produced showed, in preliminary tests, potential for use in the immunodiagnosis of human CHD [19,21].…”

Section: Introductionmentioning

confidence: 99%

A set of recombinant antigens from Echinococcus granulosus with potential for use in the immunodiagnosis of human cystic hydatid disease

Virginio

Hernández

Rott

et al. 2003

Clinical and Experimental Immunology

101

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SUMMARYSeveral recombinant clones expressing antigens from Echinococcus granulosus were isolated previously from a parasite cDNA library using cystic hydatid disease (CHD) patients' sera or rabbit hyperimmune antiserum against a lipoproteic fraction from bovine cyst fluid. Six of these antigens were expressed in Escherichia coli and the purified recombinant proteins were tested in enzyme-linked immunosorbent assay (ELISA) for specific IgG with a panel of sera from patients with surgically confirmed ( n = 58) or immunologically diagnosed ( n = 71) CHD. Sera from clinically normal individuals ( n = 203) and sera from individuals with other helminthic infections ( n = 65) were assayed for the assessment of specificity. A cut-off value was determined by receiver-operating-characteristic plots for each antigen. A recombinant antigen B subunit (AgB8/2) presented the highest sensitivity (93·1%), considering the group of sera from patients with CHD surgically confirmed, and specificity (99·5%) and is proposed as the basis for an immunodiagnostic test. The other recombinant antigens tested presented sensitivities between 58·6% and 89·7%, and three of them were considered of complementary value. In subclass-specific ELISA, different IgG isotypes showed dominance in the response for each of the recombinant antigens. There was a clear predominance of IgG4 response for all antigens tested, indicating that this would be the subclass of choice to be assessed for these recombinant proteins.

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Molecular cloning and characterization of an Echinococcus granulosus cDNA encoding malate dehydrogenase

Cited by 15 publications

References 8 publications

Comparative Analysis of the Diagnostic Performance of Six Major Echinococcus granulosus Antigens Assessed in a Double-Blind, Randomized Multicenter Study

Comparative Analysis of the Diagnostic Performance of Six Major Echinococcus granulosus Antigens Assessed in a Double-Blind, Randomized Multicenter Study

Fructose-bisphosphate aldolase and enolase from Echinococcus granulosus: Genes, expression patterns and protein interactions of two potential moonlighting proteins

A set of recombinant antigens from Echinococcus granulosus with potential for use in the immunodiagnosis of human cystic hydatid disease

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