7014 had no shared lineages

7014 had no shared lineages. Somatic hypermutation of vaccine-stimulated B cell repertoires Seroconverters and non-seroconverters differed in the somatic mutation of expanded B cell clones at day 7 post-vaccination (mean mutation 6.4% and 4.3%, respectively, p=0.0041, t-test). D (diversity) and J (joining) genes in their genomes (Tonegawa, 1983). For decades, most monitoring of human antibody responses to infections or vaccines has been performed by serological measurements that can evaluate antibody specificities, while giving only limited insight into the underlying changes in clonal populations of B cells, or the gene rearrangements responsible for the antibodies. More recently, single-cell sorting and antibody gene cloning, as well as optimized culture systems and hybridoma generation, have given greater insight into the specificity and breadth of reactivity of the antibodies produced by influenza-specific B cells, and molecular understanding of the genes encoding such antibodies (Li et al., 2012; Wrammert et Sulforaphane al., 2011; Wrammert et al., 2008; Yu et al., 2008). High-throughput DNA sequencing methods now permit detailed monitoring of B cell repertoires in humans, and are starting to be applied extensively to the study of vaccine responses (Boyd et al., 2009; DeKosky et al., 2013; Jiang et al., 2013; Krause et al., 2011; Liao et al., 2011; Wu et al., 2011). It is largely unknown whether different people use similar antibody genes in their responses to common pathogen-associated antigens. With a few exceptions, such as the antibody responses to repetitive polysaccharide antigens (Ademokun et al., 2011; Park et al., 1996; Scott et al., 1989; Silverman and Lucas, 1991), there has been little evidence of similarity between different humans responses to most pathogens. Indeed, antibodies would themselves be expected to exert a selection pressure upon the pathogens they target, causing pathogens to avoid expressing antigens that are recognized by human antibody genes. Here, we conduct a detailed study of B cell clonal expansions in response to influenza vaccination, and use deep sequencing to identify clonal expansion signatures within a week of vaccination that correlate with the magnitude of the serological response in vaccinated individuals. Comparison of expanded clones to influenza-specific plasmablasts identified by single cell sorting from the same subjects demonstrates substantial overlap between these populations. More surprisingly, we identify convergent antibody responses to the H1N1 2009 influenza strain that are shared among different people, both in response to vaccination and infection. These results represent an example of a signature in immunoglobulin gene rearrangements specific to the pathogen that elicited them, and suggest that features of an individuals history of pathogen exposure can be identified by sequence analysis. Results Deep sequencing of rearranged IGH from the trivalent inactivated seasonal influenza vaccine response To take an overview of B cell responses induced by vaccination, we carried out deep sequencing of IGH from the peripheral blood B cells of 14 healthy young individuals vaccinated with the 2007 or 2008 trivalent inactivated seasonal influenza vaccine (TIV) (Moody et al., 2011). Seven individuals were seroconverters who raised at least a 4-fold increase in titer above baseline to 2 or more vaccine antigens as measured by ELISA against purified hemagglutinins (HA). The other 7 were non-seroconverters that failed to increase their vaccine-specific antibody to meet these criteria (Table S1) (Moody et al., 2011). Twelve replicate IGH libraries were prepared from independent genomic DNA template aliquots from cryopreserved peripheral blood mononuclear cells for each individual at each of 3 time points: pre-vaccination, day 7 and day 21 post-vaccination Rabbit Polyclonal to DGKI (Figure 1A). On average, 35,436 IGH sequences were analyzed for each individual. Sequencing depth was relatively evenly distributed across the time points with an average 11,661 IGH sequences pre-vaccination, 12,200 at day 7 and 11,564 at day 21. Open in a separate window Figure 1 Quantitation of clonal B cells in Sulforaphane Sulforaphane the blood following vaccination predicts seroconversion(A) Sulforaphane Replicate IGH libraries were generated from peripheral blood B cells for 14 individuals (Table S1) at three time points:.