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Updated: Jul 19, 2026
A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations
Published on: October 25, 2018
Quantitation of rare memory B cell populations by two independent and complementary approaches
1Vaccine and Gene Therapy Institute, Oregon Health and Science University, 505 NW 185th Avenue, Beaverton, OR 97006, USA.
Insights
Quantifying memory B cells (MBC) is crucial for immunology. New flow cytometry and ELISA methods enable precise, multi-antigen MBC detection in clinical samples, improving immunological research.
Area of Science:
- Immunology
- Cellular immunology
- B cell biology
Background:
- Current methods for quantifying memory B cells (MBC) are limited.
- Existing assays often fail to detect multiple MBC specificities simultaneously.
Purpose of the Study:
- To develop novel, independent approaches for antigen-specific MBC quantitation.
- To enable simultaneous detection of multiple MBC specificities in clinical samples.
Main Methods:
- Development of a sensitive flow cytometry (FC) assay for quantifying MBC specific to tetanus and diphtheria antigens.
- Establishment of an ELISA-based MBC limiting dilution assay (LDA) for analyzing 8-12 different MBC frequencies from single blood samples.
Main Results:
- Significant correlation found between MBC numbers measured by FC and frequencies obtained by LDA (R(2)=0.92, p=0.0002).
- Both methods provide accurate quantitation of rare MBC populations.
Conclusions:
- Two distinct, validated methods (FC and LDA) are now available for antigen-specific MBC quantitation.
- These approaches are suitable for analyzing MBCs specific to simple or complex antigens and applicable across various model systems.
Abstract:
Current methodology for quantitation of memory B cells (MBC) in clinical samples is limited and often does not allow for detection of multiple MBC specificities in a single assay. Here we describe two independent approaches to antigen-specific MBC quantitation. First, a sensitive flow cytometry (FC) assay was developed for simultaneous quantitation of two prototypical B cell antigens, tetanus and diphtheria. Second, an ELISA-based MBC limiting dilution assay (LDA) was developed that provides quantitative analysis of up to 8-12 different MBC frequencies from a single blood sample. Cross-validation studies indicated that MBC numbers measured by FC correlated significantly with the frequencies obtained by LDA (R(2)=0.92, p=0.0002). These two functionally distinct approaches will be useful for accurate quantitation of rare MBC populations specific for either simple antigens (e.g. tetanus and diphtheria) or complex antigens (e.g. vaccinia or other viruses), and is amenable for use in a variety of model systems.
