title: Serum antibody responses to Wolbachia surface protein in patients with human lymphatic filariasis.
abstract: Wolbachia surface protein (WSP), which is the most abundantly expressed protein of Wolbachia from the human filarial parasite Brugia malayi, was chosen for the present study. B-cell epitope prediction of the WSP protein sequence indicates a high antigenicity, surface probability and hydrophilicity by DNA STAR software analysis. ProPred analysis suggests the presence of HLA class II binding regions in the WSP protein that contribute to T-cell responses and isotype reactivity. In order to validate these findings, the gene coding for endosymbiont WSP was PCR-amplified from the genomic DNA of the human filarial parasite Brugia malayi and cloned in T-7 expression vector pRSET-A. Western blot and ELISA at the total IgG level with recombiant WSP indicated a significantly elevated reactivity in CP compared to MF, EN and NEN individuals. Isotype ELISA also suggested an elevated reactivity in CP patients at the IgG1 level. In contrast, WSP-specific IgG4 levels were found to be elevated in MF patients compared to CP and EN. Besides this, WSP-specific IgE levels indicated an elevated reactivity in CP and MF patients compared to normals. Observations from ELISA supported the in silico predictions that indicate the presence of B- and T-cell epitopes. Hence, a combinatorial approach of in silico predictions and wet-lab studies provides interesting insights into the role of Wolbachia proteins in filarial pathogenesis.
authors: Centre for Biotechnology, Anna University, Tamil Nadu, India.
authors: Chandanapurath
authors: C
authors: Shiny
authors: Nagampalli S A
authors: NS
authors: Krushna
authors: Bairavasundaram
authors: B
authors: Archana
authors: Begum
authors: B
authors: Farzana
authors: Rangarajan B
authors: RB
authors: Narayanan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000097:blood*; D001402:B-Lymphocytes / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology; D017178:Brugia malayi / Q000382:microbiology*; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000235:genetics; D004605:Elephantiasis, Filarial / Q000097:blood / Q000276:immunology / Q000382:microbiology; D000939:Epitopes / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D006681:HLA-D Antigens / Q000276:immunology; D015801:Helminth Proteins / Q000235:genetics; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood; D007074:Immunoglobulin G / Q000097:blood; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology; D016133:Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000276:immunology; D020577:Wolbachia / Q000276:immunology*
chemical_list: D000906:Antibodies; D001426:Bacterial Proteins; D016054:DNA, Protozoan; D000939:Epitopes; D006681:HLA-D Antigens; D015801:Helminth Proteins; D007074:Immunoglobulin G; D008565:Membrane Proteins; D007073:Immunoglobulin E