Analysis of Antibody Response in Human Dengue Patients from the Mexican Coast Using Recombinant Antigens

Abstract

This study was undertaken to evaluate the feasibility of using recombinant dengue proteins to discriminate between acute dengue infections versus uninfected dengue samples. Dengue virus proteins E, NS1, NS3, and NS4B were cloned as fusion proteins and expressed in Escherichia coli. Recombinant products were tested in 100 serum samples obtained from acute dengue fever cases collected from 3 states of Mexico where dengue is endemic. Sera from 75 healthy individuals living in nonendemic areas for dengue were used as a control group. In sera from the dengue patients group, antibody responses to E protein were demonstrated in 91% of cases and NS1 protein was recognized to various extents (99%) within the first 7 days of infection. The antibody responses to NS3 and NS4B were frequently of low magnitude. Consistent negative antibody responses to all proteins were found in sera from the control group. These data suggest that the glutathione-S-transferase (GST)-dengue fusion proteins may be feasible antigens for a sensitive and specific serological assay.

Development and clinical evaluation of a highly accurate dengue NS1 rapid test: from the preparation of a soluble NS1 antigen to the construction of an RDT

“…In fact, protein yield (40 mg/l) was also not poor. It was comparable (10-30 mg/l of Huang et al) or better (8-12 mg/ml of Lazaro-Olan et al) than previous reports, in which the same E. coli system is used (Huang, et al, 2001;Lazaro-Olan, et al, 2008). Recently, functionally active DENV2 NS1 recombinant protein was successfully expressed and purified without any in vitro posttranslational modification; that is, a soluble NS1 protein was purified under the native condition (Sankar, et al, 2013).…”

“…However, most attempts to express DENV NS1 proteins in the bacterial expression system (Escherichia coli) resulted in insoluble proteins (Amorim, et al, 2010;Das, Mongkolaungkoon, & Suresh, 2009;Huang, et al, 2001;Lazaro-Olan, et al, 2008;Sankar, et al, 2013), requiring denaturation and refolding to harvest proteins from inclusion bodies.…”

“…Since the study aims to generate mAbs highly specific for the DENV NS1 protein, the priority considered in protein expression and purification was to obtain a natively antigenic protein at high purity rather than to produce protein with high yield, which many researches have focused on for years (Amorim, et al, 2010;Das, et al, 2009;Huang, et al, 2001;Lazaro-Olan, et al, 2008;Sankar, et al, 2013). High purity of the DENV NS1 protein was successfully achieved by 2-step purification using ion exchange and affinity chromatographic techniques (Fig.…”

Antibody response to dengue virus

“…1) seem to be involved in both protection and pathogenesis. Immune responses have also been shown to target other non-structural proteins, albeit with reduced intensity and frequency [23].…”

Clonagem e expressão das proteínas recombinantes NS1 e NS3 do vírus da dengue tipo 3

“…Por outro lado, a proteína NS3 desencadeia uma forte resposta imune celular e uma fraca resposta humoral, o que poderia explicar a ausência de detecção pelo soro do paciente número 4 (BRINTON et al, 1998;LAZARO-OLAN et al, 2008;MATHEW;ROTHMAN, 2008;ROTHMAN, 2004).…”