A clinical microchip for evaluation of single immune cells reveals high functional heterogeneity in phenotypically similar T cells
Related Experiment Video
Updated: May 5, 2026
Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations
Published on: January 19, 2017
Chao Ma1, Rong Fan, Habib Ahmad
1NanoSystems Biology Cancer Center, California Institute of Technology, Pasadena, California, USA.
Insights
This study introduces a microfluidic platform for analyzing single-cell immune responses. It reveals significant functional diversity in tumor-specific T cells, aiding immune monitoring.
Area of Science:
- Immunology
- Biotechnology
- Microfluidics
Background:
- Cellular immunity exhibits high functional heterogeneity.
- Analyzing diverse effector molecules from single cells is crucial for understanding immune responses.
- Existing methods may lack the multiplexing or efficiency needed for comprehensive analysis.
Purpose of the Study:
- To develop and validate a microfluidic platform for highly multiplexed, quantitative single-cell protein analysis.
- To assess the functional heterogeneity of tumor antigen-specific cytotoxic T lymphocytes (CTLs).
- To compare the platform's performance against standard immunotechnologies.
Main Methods:
- A novel microfluidic platform enabling measurement of >10 secreted proteins per cell.
- Validation using lipopolysaccharide (LPS)-stimulated human macrophages and cytokine analysis.
- Ex vivo analysis of T cell polyfunctional diversity in tumor patients versus healthy donors.
Main Results:
- The platform reliably quantifies multiple secreted proteins from single cells with high sample efficiency.
- Profound, yet focused, functional heterogeneity was observed in tumor antigen-specific CTLs.
- Major functional phenotypes of responding T cells were quantitatively identified.
Conclusions:
- The developed microfluidic platform is an informative tool for immune monitoring.
- It enables quantitative assessment of T cell polyfunctional diversity.
- This technology can advance clinical assessment and understanding of immune responses.
Abstract:
Cellular immunity has an inherent high level of functional heterogeneity. Capturing the full spectrum of these functions requires analysis of large numbers of effector molecules from single cells. We report a microfluidic platform designed for highly multiplexed (more than ten proteins), reliable, sample-efficient (∼1 × 10(4) cells) and quantitative measurements of secreted proteins from single cells. We validated the platform by assessment of multiple inflammatory cytokines from lipopolysaccharide (LPS)-stimulated human macrophages and comparison to standard immunotechnologies. We applied the platform toward the ex vivo quantification of T cell polyfunctional diversity via the simultaneous measurement of a dozen effector molecules secreted from tumor antigen-specific cytotoxic T lymphocytes (CTLs) that were actively responding to tumor and compared against a cohort of healthy donor controls. We observed profound, yet focused, functional heterogeneity in active tumor antigen-specific CTLs, with the major functional phenotypes quantitatively identified. The platform represents a new and informative tool for immune monitoring and clinical assessment.
More Related Videos
09:34A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations
Published on: October 25, 2018
08:02Determination of Immune Cell Identity and Purity Using Epigenetic-Based Quantitative PCR
Published on: February 19, 2020
