期刊
CLINICAL IMMUNOLOGY
卷 129, 期 1, 页码 10-18出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.clim.2008.06.009
关键词
human; microengraving; type 1 diabetes; single cell analysis; autoantibodies; cytokines; proinsulin
类别
资金
- NIH Autoimmunity Prevention Center
- Broad Institute of MIT
- Harvard
- Juvenile Diabetes Research Foundation International [NIH Al651003]
- National Institutes of Health [R01 Al39229, R01 Al44447, P01 Al39671]
- Jacob Javits Merit Award [NS2427]
- National Institute of Neurological Disorders and Stroke
- National Multiple Sclerosis Society
Cell surface determinants, cytokines and antibodies secreted by hematopoietic cells are used to classify their lineage and function. Currently available techniques are unable to elucidate multiple secreted proteins white also assigning phenotypic surface-displayed markers to the individual living cells. Here, a soft lithographic method, microengraving, was adapted for the multiplexed interrogation of populations of individual human peripheral blood mononuclear cells for secreted cytokines (IFN-gamma and IL-6), antigen-specific antibodies, and lineage-specific surface-expressed markers. Application of the method to a clinical sample from a recent-onset Type 1 diabetic subject with a positive titer of anti-insulin antibodies showed that similar to 0.58% of circulating CD19(+) B cells secreted proinsulin-reactive antibodies of the IgG isotype and 2-3% of circulating cells secreted IL-6. These data demonstrate the utility of microengraving for interrogating multiple phenotypes of single human cells concurrently and for detecting rare populations of cells by their secreted products. (C) 2008 Elsevier Inc. All rights reserved.
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