1. Identification and characterization of the Onchocerca volvulus Excretory Secretory Product Ov28CRP, a putative GM2 activator protein
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Philippe Poelvoorde, Lea Olive Tchouate Gainkam, Arnaud Poterszman, Ferdinand Ngale Njume, Stephen Mbigha Ghogomu, Evelina Edelweiss, Robert Adamu Shey, Susi Anheuser, Christophe Lelubre, Annie Robert, Luc Vanhamme, Joseph Kamgno, Leon Mutesa, Jacob Souopgui, Perrine Humblet, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), Laboratory of Molecular Parasitology (IBMM), Université libre de Bruxelles (ULB), Pôle Epidémiologie et Biostatistique, Université Catholique de Louvain = Catholic University of Louvain (UCL)-Institut de Recherche Expérimentale et Clinique (IREC)-Faculté de Santé Publique (FSP), Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Center for Microscopy and Molecular Imaging (IBMM - CMMI), Institut de Biologie et de Médecine Moléculaires [Gosselies] (ULB/IBMM), Faculté des Sciences [Bruxelles] (ULB), Université libre de Bruxelles (ULB)-Université libre de Bruxelles (ULB)-Faculté de Médecine [Bruxelles] (ULB), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and UCL - SSS/IREC/EPID - Pôle d'épidémiologie et biostatistique
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0301 basic medicine ,Male ,Nematoda ,[SDV]Life Sciences [q-bio] ,RC955-962 ,Onchocerciasis ,Santé publique ,law.invention ,0302 clinical medicine ,law ,Arctic medicine. Tropical medicine ,Sf9 Cells ,Medicine and Health Sciences ,Recombinant Protein Purification ,Cloning, Molecular ,Enzyme-Linked Immunoassays ,Nematode Infections ,Pathogen ,Immune Response ,ComputingMilieux_MISCELLANEOUS ,medicine.diagnostic_test ,integumentary system ,Eukaryota ,Helminth Proteins ,DNA, Helminth ,Recombinant Proteins ,3. Good health ,[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM] ,Infectious Diseases ,Helminth Infections ,Recombinant DNA ,Female ,Onchocerca ,Public aspects of medicine ,RA1-1270 ,Research Article ,Neglected Tropical Diseases ,Signal peptide ,Sequence analysis ,Protein Purification ,030231 tropical medicine ,Immunology ,Biology ,Spodoptera ,Research and Analysis Methods ,Microbiology ,Host-Parasite Interactions ,03 medical and health sciences ,Western blot ,Onchocerciasis, Ocular ,Helminths ,parasitic diseases ,medicine ,Parasitic Diseases ,Animals ,Humans ,Immunoassays ,Gene ,G(M2) Activator Protein ,Gene Expression Profiling ,Public Health, Environmental and Occupational Health ,Insect cell culture ,Organisms ,Biologie moléculaire ,Biology and Life Sciences ,Sequence Analysis, DNA ,biology.organism_classification ,Tropical Diseases ,Onchocerca volvulus ,Invertebrates ,030104 developmental biology ,Immunoglobulin G ,Immunologic Techniques ,Cattle ,Sciences pharmaceutiques ,Purification Techniques - Abstract
Onchocerca volvulus is the nematode pathogen responsible for human onchocerciasis also known as “River blindness”, a neglected tropical disease that affects up to 18 million people worldwide. Helminths Excretory Secretory Products (ESPs) constitute a rich repertoire of molecules that can be exploited for host-parasite relationship, diagnosis and vaccine studies. Here, we report, using a range of molecular techniques including PCR, western blot, recombinant DNA technology, ELISA, high performance thin-layer chromatography and mass spectrometry that the 28 KDa cysteine-rich protein (Ov28CRP) is a reliable component of the O. volvulus ESPs to address the biology of this parasite. We showed that (1) Ov28CRP is a putative ganglioside GM2 Activator Protein (GM2AP) conserved in nematode; (2) OvGM2AP gene is transcriptionally activated in all investigated stages of the parasitic life cycle, including larval and adult stages; (3) The full-length OvGM2AP was detected in in-vitro O. volvulus ESPs of adult and larval stages; (4) the mass expressed and purified recombinant OvGM2AP purified from insect cell culture medium was found to be glycosylated at asparagine 173 and lacked N-terminal signal peptide sequence; (5) the recombinant OvGM2AP discriminated serum samples of infected and uninfected individuals; (6) OvGM2AP competitively inhibits MUG degradation by recombinant β-hexosaminidase A but not MUGS, and could not hydrolyze the GM2 to GM3; (7) humoral immune responses to the recombinant OvGM2AP revealed a negative correlation with ivermectin treatment. Altogether, our findings suggest for the first time that OvGM2AP is an antigenic molecule whose biochemical and immunological features are important to gain more insight into our understanding of host-parasite relationship, as well as its function in parasite development at large., SCOPUS: ar.j, info:eu-repo/semantics/published
- Published
- 2019
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