Molecular cloning and serological characterization of a Brugia malayi pepsin inhibitor homolog

Dissanayake, Senarath; Xu, Min; Nkenfou, Céline Nguefeu; Piessens, Willy F.

doi:10.1016/0166-6851(93)90191-y

Cited by 33 publications

(38 citation statements)

References 6 publications

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“…Serum antibody specific for Ov33-3 was detected in patients infected with O. volvulus (25,35). Antibodies against Bm33 have been found in B. malayi-infected patients (12). Serum antibodies specific for DiT33 were found in D. immitis-infected dogs (21,33) and cats (18,33).…”

Section: Discussionmentioning

confidence: 99%

An Aspartyl Protease Inhibitor Orthologue Expressed byParelaphostrongylus tenuisIs Immunogenic in an Atypical Host

Duffy

MacAfee

Burt

et al. 2002

Clin Vaccine Immunol

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Parelaphostrongylus tenuis is a neurotropic nematode common in white-tailed deer (Odocoileus virginianus) of eastern North America. This parasite is the causative agent of a debilitating neurologic disease in atypical hosts, including domestic livestock. In order to identify proteins of potential significance in the host-parasite relationship, a cDNA library was produced from adult P. tenuis mRNA. Screening the library with antisera from infected red deer (Cervus elaphus elaphus) and immunized AO strain rats, we identified clones with sequence similarities to aspartyl protease inhibitors from several parasitic nematodes. Antibody that was generated against this recombinant protein of P. tenuis (Pt-API-1) detected the native protein in E/S products, in muscle and gonad, and on the surface of the cuticle of adult male and female P. tenuis. The native protein was detected in internal structures of first-stage (L1) and third-stage (L3) larvae. Reverse transcription-PCR confirmed expression of Pt-api-1 in L1, L3, and adult male and female worms. Expression of Pt-API-1 throughout the life cycle of P. tenuis suggests an essential function. Antibodies specific for recombinant Pt-API-1 were detected by enzyme-linked immunosorbent assay in sera from 12 red deer experimentally infected with P. tenuis. Antibodies were detected within 28 to 56 days postinfection. Responses were sustained or biphasic in animals with patent infections, consistent with expression of Pt-API-1 by L1. Our results are compatible with findings in other parasitic nematodes showing that aspartyl protease inhibitors are highly immunogenic.

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Section: Discussionmentioning

confidence: 99%

An Aspartyl Protease Inhibitor Orthologue Expressed byParelaphostrongylus tenuisIs Immunogenic in an Atypical Host

Duffy

MacAfee

Burt

et al. 2002

Clin Vaccine Immunol

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“…Equimolar concentrations of Bm-Aspin and the proteases were mixed and incubated in 100 mM sodium acetate buffer at pH 5.6 for 10 min at 37°C separately. Bm-Aspin initially termed as “pepsin inhibitor homologue” [16] based on the sequence homology with Ascaris Suum PI-3 and was believed to follow similar kind of pepsin inhibition. Hence pH 5.6 was considered.…”

Section: Methodsmentioning

confidence: 99%

“…In this context, an Aspin from Brugia malayi , termed as pepsin inhibitor homolog compared to the amino acid composition of PI-3 from Ascaris Suum [16], has been characterized as an immunodominant and hypodermal antigen [17]–[18]. In addition to this, we have recently reported the pepsin inhibition activity and physicochemical characterization of Bm-Aspin [19]–[20].…”

Section: Introductionmentioning

confidence: 99%

A Structural Biology Approach to Understand Human Lymphatic Filarial Infection

et al. 2014

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The presence of aspartic protease inhibitor in filarial parasite Brugia malayi (Bm-Aspin) makes it interesting to study because of the fact that the filarial parasite never encounters the host digestive system. Here, the aspartic protease inhibition kinetics of Bm-Aspin and its NMR structural characteristics have been investigated. The overall aim of this study is to explain the inhibition and binding properties of Bm-Aspin from its structural point of view. UV-spectroscopy and multi-dimensional NMR are the experiments that have been performed to understand the kinetic and structural properties of Bm-Aspin respectively. The human aspartic proteases that are considered for this study are pepsin, renin, cathepsin-E and cathepsin-D. The results of this analysis performed with the specific substrate [Phe-Ala-Ala-Phe (4-NO2)-Phe-Val-Leu (4-pyridylmethyl) ester] against aspartic proteases suggest that Bm-Aspin inhibits the activities of all four human aspartic proteases. The kinetics studies indicate that Bm-Aspin follows a competitive mode of inhibition for pepsin and cathepsin-E, non-competitive for renin and mixed mode for cathepsin-D. The triple resonance NMR experiments on Bm-Aspin suggested the feasibility of carrying out NMR studies to obtain its solution structure. The NMR titration studies on the interactions of Bm-Aspin with the proteases indicate that it undergoes fast-exchange phenomena among themselves. In addition to this, the chemical shift perturbations for some of the residues of Bm-Aspin observed from 15N-HSQC spectra upon the addition of saturated amounts of aspartic proteases suggest the binding between Bm-Aspin and human aspartic proteases. They also provide information on the variations in the intensities and mode of binding between the proteases duly corroborating with the results from the protease inhibition assay method.

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“…To be specific, Wolbachia proteins such as Wolbachia Surface Protein (WSP), Heat Shock Protein 60 (HSP 60) decreases CD4 + T cell activation and impairs the proliferative responses in filarial patients compared to the uninfected group of individuals thus contributing to the suppression of immune responses in patients [8,9]. Further, a hypodermal protein from L3 stage of the parasite, Bm33 was identified and was characterized [10]. Bm33 was produced using recombinant technology (rBm33) and was characterized to show high levels of antigen specific IgG4 antibodies in microfilaria positive individuals compared to the patients with chronic pathology [11].…”

Section: Introductionmentioning

confidence: 99%

Recombinant Brugia malayi pepsin inhibitor (rBm33) induced monocyte function and absence of apoptotic cell death: An in vitro study

Sreenivas

Vijayan

Babu

et al. 2012

Microbial Pathogenesis

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Molecular cloning and serological characterization of a Brugia malayi pepsin inhibitor homolog

Cited by 33 publications

References 6 publications

An Aspartyl Protease Inhibitor Orthologue Expressed byParelaphostrongylus tenuisIs Immunogenic in an Atypical Host

An Aspartyl Protease Inhibitor Orthologue Expressed byParelaphostrongylus tenuisIs Immunogenic in an Atypical Host

A Structural Biology Approach to Understand Human Lymphatic Filarial Infection

Recombinant Brugia malayi pepsin inhibitor (rBm33) induced monocyte function and absence of apoptotic cell death: An in vitro study

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