Epitope-Specific Human Influenza Antibody Repertoires Diversify by B Cell Intraclonal Sequence Divergence and Interclonal Convergence

Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA.
The Journal of Immunology (Impact Factor: 4.92). 08/2011; 187(7):3704-11. DOI: 10.4049/jimmunol.1101823
Source: PubMed


We generated from a single blood sample five independent human mAbs that recognized the Sa antigenic site on the head of influenza hemagglutinin and exhibited inhibitory activity against a broad panel of H1N1 strains. All five Abs used the V(H)3-7 and J(H)6 gene segments, but at least four independent clones were identified by junctional analysis. High-throughput sequence analysis of circulating B cells revealed that each of the independent clones were members of complex phylogenetic lineages that had diversified widely using a pattern of progressive diversification through somatic mutation. Unexpectedly, B cells encoding multiple diverging lineages of these clones, including many containing very few mutations in the Ab genes, persisted in the circulation. Conversely, we noted frequent instances of amino acid sequence convergence in the Ag combining sites exhibited by members of independent clones, suggesting a strong selection for optimal binding sites. We suggest that maintenance in circulation of a wide diversity of somatic variants of dominant clones may facilitate recognition of drift variant virus epitopes that occur in rapidly mutating virus Ags, such as influenza hemagglutinin. In fact, these Ab clones recognize an epitope that acquired three glycosylation sites mediating escape from previously isolated human Abs.

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Available from: James E Crowe
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    • "This article, published in Genome Research, is available under a Creative Commons License (Attribution-NonCommercial 3.0 Unported), as described at 2008), used in numerous biological studies (Sanchez et al. 2003; Campbell et al. 2008; Boyd et al. 2009, 2010; Krause et al. 2011; Jager et al. 2012; Lev et al. 2012; Maletzki et al. 2012), and optimized for clinical use (McClure et al. 2006; Harris et al. 2012; Sproul and Goodlad 2012), although the potential for biased PCR amplification remains. The 59 rapid amplification of cDNA ends (59 RACE) has also been used (Bertioli 1997; Freeman et al. 2009; Varadarajan et al. 2011; Warren et al. 2011), but can suffer from low efficiency and high levels of nonspecific amplification, contamination by short fragments from RNA degradation, or incomplete cDNA synthesis. "
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