Format

Send to:

Choose Destination
See comment in PubMed Commons below
Vaccine. 2014 May 19;32(24):2866-73. doi: 10.1016/j.vaccine.2014.02.020. Epub 2014 Mar 3.

Profiling human antibody responses by integrated single-cell analysis.

Author information

  • 1Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, United States.
  • 2Department of Chemical and Biomolecular Engineering, University of Houston, Houston, TX 77204, United States.
  • 3International AIDS Vaccine Initiative, Scripps Research Institute, La Jolla, CA 92037, United States.
  • 4The Ragon Institute of MGH, MIT and Harvard, Cambridge, MA 02139, United States.
  • 5Department 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.

Abstract

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.

KEYWORDS:

Humoral responses; Immune profiling; Microengraving; Nanowells; Plasma and memory B cells analysis

PMID:
24602776
[PubMed - indexed for MEDLINE]
PMCID:
PMC4164152
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science Icon for PubMed Central
    Loading ...
    Write to the Help Desk