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Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry
Published on: June 26, 2018
Single-cell deep phenotyping of IgG-secreting cells for high-resolution immune monitoring
Klaus Eyer1, Raphaël C L Doineau2,3, Carlos E Castrillon4,5
1Laboratoire Colloïdes et Matériaux Divisés (LCMD), ESPCI Paris, PSL Research University, CNRS UMR8231 Chimie Biologie Innovation, Paris, France.
Insights
A new microfluidic system, DropMap, enables high-throughput analysis of individual antibody-secreting cells. This technology reveals dynamic changes in antibody secretion rate and affinity following immunization.
Area of Science:
- Immunology
- Microfluidics
- Cellular Analysis
Background:
- Analyzing individual antibody-secreting cells is crucial for understanding immune responses.
- Existing methods lack the quantitative, high-throughput capabilities needed for detailed analysis.
Purpose of the Study:
- To introduce DropMap, a novel microfluidic system for analyzing single antibody-secreting cells.
- To characterize the dynamics of antibody secretion rate and affinity in vivo.
Main Methods:
- Development of a microfluidic system (DropMap) for single-cell compartmentalization in droplets.
- Utilizing a fluorescence relocation-based immunoassay for simultaneous analysis of secretion rate and affinity.
- Characterization of over 0.5 million antibody-secreting cells from immunized mice.
Main Results:
- DropMap enables analysis of antibody-secreting cells in tens of thousands of droplets.
- Immunization with tetanus toxoid (TT) significantly increased the range of single-cell secretion rates (3 logs) and affinities (4 logs).
- Observed dynamic differences in secretion rate and affinity over time and across anatomical compartments.
Conclusions:
- The DropMap system provides a powerful tool for quantitative, high-throughput analysis of antibody-secreting cells.
- This technology can optimize immunization and vaccination protocols and advance antibody screening.
- Reveals dynamic immune response characteristics at the single-cell level.
Abstract:
Studies of the dynamics of the antibody-mediated immune response have been hampered by the absence of quantitative, high-throughput systems to analyze individual antibody-secreting cells. Here we describe a simple microfluidic system, DropMap, in which single cells are compartmentalized in tens of thousands of 40-pL droplets and analyzed in two-dimensional droplet arrays using a fluorescence relocation-based immunoassay. Using DropMap, we characterized antibody-secreting cells in mice immunized with tetanus toxoid (TT) over a 7-week protocol, simultaneously analyzing the secretion rate and affinity of IgG from over 0.5 million individual cells enriched from spleen and bone marrow. Immunization resulted in dramatic increases in the range of both single-cell secretion rates and affinities, which spanned at maximum 3 and 4 logs, respectively. We observed differences over time in dynamics of secretion rate and affinity within and between anatomical compartments. This system will not only enable immune monitoring and optimization of immunization and vaccination protocols but also potentiate antibody screening.
