Isolation and Characterization of Antigen-Specific Plasmablasts Using a Novel Flow Cytometry-Based Ig Capture Assay.
Isolation and Characterization of Antigen-Specific Plasmablasts Using a Novel Flow Cytometry-Based Ig Capture Assay.
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DOI：
10.4049/jimmunol.1701253
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发表时间：
2017-12-15
期刊：
影响因子：
--
通讯作者：
McKay PF
中科院分区：
文献类型：
--
作者：
Pinder CL;Kratochvil S;Cizmeci D;Muir L;Guo Y;Shattock RJ;McKay PF
We report the development of a novel flow cytometry-based immunoglobulin capture assay (ICA) for the identification and sorting of individual antibody secreting cells based on their antigen reactivity. The ICA represents a fast and versatile tool for single-cell sorting of peripheral plasmablasts, streamlining subsequent antibody analysis and cloning. We demonstrate the utility of the assay by isolating antigen-reactive plasmablasts from cryopreserved PBMC obtained from volunteers vaccinated with a recombinant HIV envelope protein. To show the specificity of the ICA, we produced antigen-specific antibodies from these cells and subsequently verified their antigen reactivity via ELISA. Furthermore, we used the ICA to track antigen-specific plasmablast responses in HIV-vaccine recipients over a period of 42 days and performed a head-to-head comparison with a conventional B-cell ELISpot. Results were highly comparable, highlighting that this assay is a viable alternative for monitoring antigen-specific plasmablast responses at early time points after infection or vaccination. The ICA provides important added benefits in that phenotypic information can be obtained from the identified antigen-specific cells that can then be captured for downstream applications such as B cell sequencing and/or antibody cloning. We envisage the ICA as being a useful tool in antibody repertoire analysis for future clinical trials.
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发表时间：
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