Nonstructural protein NS1 immunodominant epitope detected specifically in dengue virus infected material by a SELDI‐TOF/MS based assay

Abstract

Dengue virus (DV) infection is the most common mosquito-born viral disease of public health significance. Though most patients only suffer from flu-like symptoms, a small group of patients experiences more severe forms of the disease. The viral nonstructural protein 1 (NS1), a secreted protein correlating with viremia, is a key element used for dengue diagnosis with potential implications in severe dengue prognosis. Capture-ELISAs for the early detection of the NS1 protein in the sera during the acute febrile stage are commonly used in routine by diagnostic laboratories. In this study, the detection of NS1 protein in DV-infected material was assessed by an alternative method combining a single NS1-directed monoclonal antibody and the SELDI-TOF/MS technology. According to the epitope mapping, the antibodies used are mainly directed against an immuno-dominant peptide located on the C-terminal part of the protein. The NS1 SELDI-TOF assay is specific, has a sensitivity level close to capture-ELISAs and is potentially useful for a coupled serotyping/detection assay or for the detection of subtle post-translational modifications on the protein.

Differential proteomic analysis of virus-enriched fractions obtained from plasma pools of patients with dengue fever or severe dengue

“…HepG2 cells (ATCC HB-8065) were cultivated at 37 °C in 5 % CO 2 in DMEM supplemented with 10 % decomplemented fetal calf serum, 5 × 10 4 IU Penicillin, 50 mg streptomycin and 10 mM L- glutamine (Invitrogen, Paisley, UK). The cells were infected as previously described [ 22 ] with a serotype 3 DV (DV3, strain D78-878 Thailand) graciously provided by Dr V. Barban (Sanofi-Pasteur, Lyon, France). Sub-confluent HepG2 cell cultures (approx.…”

A Highly Conserved Region Between Amino Acids 221 and 266 of Dengue Virus Non-Structural Protein 1 is a Major Epitope Region in Infected Patients

“…This epitope region has not been reported previously, which may be because of the different origins of the MAbs used in previous studies. 10,[29][30][31][32][33][34][35][36][37] Here, we performed epitope mapping using HuMAbs derived from DENV-infected patients, whereas previous studies used mouse MAbs. According to the results of the previous studies, almost the entire NS1 region could act as an epitope region recognized by mouse MAbs, whereas HuMAbs map a limited region located between amino acids 221 and 266.…”

“…10,[29][30][31][32][33][34][35][36][37][38][39] These studies hypothesized that anti-NS1 antibodies cross-react with host molecules and subsequently, induce plasma leakage. Some mouse anti-NS1 MAbs crossreact with host molecules that share similar epitopes with NS1.…”

Comprehensive mapping infection-enhancing epitopes of dengue pr protein using polyclonal antibody against prM

“…Epitopes are the focus of pathogenesis research as well as the development of vaccine and diagnostic reagent (Wu et al 2001 , 2003 ; Jiang et al 2010 ). Epitopes in the E (Li et al 2012 ; de Alwis et al 2012 ; Lin et al 2012 ; Hughes et al 2012 ; Wahala et al 2012 ), NS1( Wu et al 2001 , 2003 ; Jiang et al 2010 ; Steidel et al 2012 ), NS4a, and C (Anandarao et al 2005 ) have been well mapped. Although there have been attempts to locate the epitopes of prM (Falconar 1999 ; Huang et al 2008 ; Song et al 2013 ; Luo et al 2013 ), the functional roles of the prM protein in DENV infection and the precise antigenic structures of prM are not yet fully understood.…”