Profiling Human Antibody Responses by Integrated Single-Cell Analysis

dc.contributor.authorOgunniyi, Adebola O.
dc.contributor.authorThomas, Brittany A.
dc.contributor.authorPolitano, Timothy J.
dc.contributor.authorVaradarajan, Navin
dc.contributor.authorLandais, Elise
dc.contributor.authorPoignard, Pascal
dc.contributor.authorWalker, Bruce D.
dc.contributor.authorKwon, Douglas S.
dc.contributor.authorLove, Christopher J.
dc.description.abstractComprehensive characterization of the antigen-specific B cells induced during infections or following vaccination would facilitate the discovery of novel antibodies and inform how interventions shape protective humoral responses. The analysis of human B cells and their antibodies has been performed using flow cytometry to evaluate memory B cells and expanded plasmablasts, while microtechnologies have also provided a useful tool to examine plasmablasts/plasma cells after vaccination. Here we present an integrated analytical platform, using arrays of subnanoliter wells (nanowells), for constructing detailed profiles for human B cells comprising the immunophenotypes of these cells, the distribution of isotypes of the secreted antibodies, the specificity and relative affinity for defined antigens, and for a subset of cells, the genes encoding the heavy and light chains. The approach combines on-chip image cytometry, microengraving, and single-cell RT-PCR. Using clinical samples from HIV-infected subjects, we demonstrate that the method can identify antigen-specific neutralizing antibodies, is compatible with both plasmablasts/plasma cells and activated memory B cells, and is well-suited for characterizing the limited numbers of B cells isolated from tissue biopsies (e.g., colon biopsies). The technology should facilitate detailed analyses of human humoral responses for evaluating vaccines and their ability to raise protective antibody responses across multiple anatomical compartments.
dc.identifier.citationCopyright 2014 Vaccine. This is a post-print version of a published paper that is available at: Recommended citation: Ogunniyi, Adebola O., Brittany A. Thomas, Timothy J. Politano, Navin Varadarajan, Elise Landais, Pascal Poignard, Bruce D. Walker, Douglas S. Kwon, and J. Christopher Love. "Profiling human antibody responses by integrated single-cell analysis." Vaccine 32, no. 24 (2014): 2866-2873. doi: 10.1016/j.vaccine.2014.02.020. This item has been deposited in accordance with the publisher copyright and licensing terms and with the author's permission.
dc.subjecthumoral responses
dc.subjectimmune profiling
dc.subjectplasma and memory B cells analysis
dc.titleProfiling Human Antibody Responses by Integrated Single-Cell Analysis


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