Profiling human antibody responses by integrated single-cell analysis.

Autor: Ogunniyi AO; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, United States., Thomas BA; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, United States., Politano TJ; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, United States., Varadarajan N; Department of Chemical and Biomolecular Engineering, University of Houston, Houston, TX 77204, United States., Landais E; International AIDS Vaccine Initiative, Scripps Research Institute, La Jolla, CA 92037, United States., Poignard P; International AIDS Vaccine Initiative, Scripps Research Institute, La Jolla, CA 92037, United States., Walker BD; The Ragon Institute of MGH, MIT and Harvard, Cambridge, MA 02139, United States., Kwon DS; The Ragon Institute of MGH, MIT and Harvard, Cambridge, MA 02139, United States., Love JC; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, United States; The Ragon Institute of MGH, MIT and Harvard, Cambridge, MA 02139, United States. Electronic address: clove@mit.edu.
Jazyk: angličtina
Zdroj: Vaccine [Vaccine] 2014 May 19; Vol. 32 (24), pp. 2866-73. Date of Electronic Publication: 2014 Mar 03.
DOI: 10.1016/j.vaccine.2014.02.020
Abstrakt: Comprehensive 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.
(Copyright © 2014 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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