期刊
MAGNETOCHEMISTRY
卷 7, 期 7, 页码 -出版社
MDPI
DOI: 10.3390/magnetochemistry7070092
关键词
CyTOF; mass cytometry; magnetic enrichment; antigen-specific B cells; antigen-binding B cells; antigen-reactive B cells; B lymphocytes; B cells
资金
- University of Colorado School of Medicine Human Immunology & Immunotherapy Initiative (HI3)
- NIH [P30 DK116073, K01 OD028759, DP3 DK110845]
This study presents a method utilizing biotinylated antigens combined with magnetic nanoparticles to detect and enrich antigen-binding B cells from peripheral blood and tissues. The CyTOF-compatible approach allows for multiplexed enrichment of antigen-reactive B cells, such as insulin and tetanus toxoid, improving monitoring of their phenotype and function while minimizing sample and run times needed.
Over the years, various techniques have been utilized to study the function and phenotype of antigen-binding B cells in the primary repertoire following immunization, infection, and development of autoimmunity. Due to the low frequency of antigen-reactive B cells (<0.05% of lymphocytes) in the periphery, preliminary enrichment of cells is necessary to achieve sufficient numbers for statistically sound characterization, especially when downstream analytic platform use, e.g., CyTOF, is low throughput. We previously described a method to detect and enrich antigen-reactive B cells from peripheral blood and tissues using biotinylated antigens in conjunction with magnetic nanoparticles, preparative to a downstream analysis by ELISPOT and flow cytometry. While mass cytometry (CyTOF) enables high dimensional immunophenotyping of over 40 unique parameters on a single-cell level, its low throughput compared to flow cytometry and requirement for removal of metal contaminants, such as nanoparticles, made it particularly unsuitable for studies of rare cells in a mixed population. Here we describe a novel CyTOF-compatible approach for multiplexed enrichment of antigen-reactive B cells, e.g., insulin and tetanus toxoid, using cleavable magnetic nanoparticles. This method allows improved monitoring of the phenotype and function of antigen-reactive B cells during the development of disease or after immunization while minimizing the amount of sample and run times needed.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据