October 9 - 12, 2012
Building 10 & Natcher Conference Center
- General Schedule of Events
- Opening Plenary Session
- Concurrent Symposia Sessions
- Poster Sessions
- National Graduate Student Research Conference
- FARE Award Ceremony
- Special Exhibits on Resources for Intramural Research
- TSA Research Festival Exhibit Show
- Clinical Center Tours
- Research Festival Committees
- Past Research Festivals
Development of a Luciferase Immunoprecipitation Systems (LIPS) Assay to Detect IgG Antibodies against Human Respiratory Syncytial Virus Nucleoprotein
| Friday, November 08, 2013 — Poster Session IV | |||
|---|---|---|---|
2:00 p.m. – 4:00 p.m. |
FAES Academic Center (Upper-Level Terrace) |
FDA/CBER |
VIROL-4 |
Authors
- S. Kumari
- R.L. Crim
- A. Kulkarni
- S. Audet
- T. Mdluli
- H. Murata
- J. Beeler
Abstract
The nucleoprotein of respiratory syncytial virus (RSV-N) is immunogenic and elicits an IgG response following infection suggesting that anti-RSV-N antibodies may be a marker of RSV exposure. The gene for RSV-N was cloned into a mammalian expression vector, pREN2, and the expressed luciferase-tagged RSV-N protein (Ruc-N) used to develop an assay to detect anti-RSV-N specific IgG antibodies based on a high-throughput immunoprecipitation method (LIPS-N-RSV). Assay specificity was evaluated using monoclonal antibodies and pre- and post-infection/immunization serum samples from rabbits, chimpanzees, and humans with proven RSV infection. Pre-immune sera and sera from rabbits immunized with paramyxoviruses were negative when tested by LIPS-N-RSV while sera from rabbits immunized with either RSV subtype A or B gave positive signals in a dose dependent manner. An anti-RSV-N MAb was positive in the LIPS-N-RSV assay while MAbs against other myxo- and paramyxoviruses nucleoprotein and against RSV-F and -G were not. Sera from chimpanzees simultaneously immunized with vaccinia recombinant viruses expressing RSV-F and -G were negative when tested using LIPS-N-RSV, however, anti-RSV-N IgG responses were detected following live RSV challenge. LIPS-N-RSV detects RSV-N specific IgG responses that may be a useful marker of infection following RSV exposure among RSV naïve populations.
