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Epitope mapping of a monoclonal antibody specific for human herpesvirus 6 variant A immediate-early 2 protein
1Laboratory of Virology, Rheumatology and Immunology Research Center, CHUQ Research Center, Quebec, Que., Canada.
Background:
Human herpesvirus 6 (HHV-6) variants A and B are distinct viruses that differ in their biological properties and association to disease. Diagnostic tools able to discriminate between these two variants and between active or latent HHV-6 infection are much needed. In our effort to develop variant-specific antibodies against HHV-6 immediate-early (IE) proteins, we had previously generated P6H8, a monoclonal antibody (mAb) reacting specifically with the immediate-early 2 (IE2) proteins from HHV-6A.
Objectives:
To characterize the P6H8 HHV-6 variant specific mAb and evaluate its potential as part of a variant-specific diagnostic tool for HHV-6A infection. Consequently, our objective was to map the epitope recognized by P6H8.
Study Design:
In order to map P6H8 reactivity by Western blotting, we generated deletion mutants of IE2 protein as well as various GST-IE2 fusion proteins. HHV-6A infected cells were used to demonstrate P6H8 reactivity against native IE2. A synthetic peptide corresponding to the P6H8 epitope was used to confirm our results and block P6H8 reactivity.
Results:
We mapped the P6H8 epitope to amino acids 1078-1089 of HHV-6A IE2. A peptide (FTPFYYQSSRTR) recreating this epitope was effective in blocking the recognition of both native and recombinant IE2 by P6H8.
Conclusions:
Our work provides a precise characterization of the P6H8 mAb and its specificity toward the IE2 protein of HHV-6 variant A which could prove useful for the differential diagnostic of active infection by HHV-6.
