期刊
NATURE MEDICINE
卷 17, 期 6, 页码 738-U133出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nm.2375
关键词
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资金
- Benjamin M. Rosen Fellowship
- US National Institutes of Health [1 K99 CA136759-01, CA-16042, AI-28697]
- US National Cancer Institute [5U54 CA119347]
- Ivy Foundation
- Jean Perkins Foundation
- California Institute for Regenerative Medicine New Faculty [RN2-00902-1]
- Caltech/UCLA Joint Center for Translational Medicine
- Melanoma Research Alliance
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 (similar to 1 x 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.
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