Assigning immunoglobulin class from single-cell transcriptomes in IgA1-secreting versus membrane subpopulations
| Autor: | Colin Reily, Nuo Xu, David K Crossman |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 70, Iss 2, Pp 89-99 (2021) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/btn-2020-0044 |
| Popis: | IgA nephropathy (IgAN) is an autoimmune disease characterized by renal glomerular immunodeposits enriched for galactose-deficient IgA1 (Gd-IgA1; autoantigen) with the corresponding IgG autoantibodies. Despite the known contribution of Gd-IgA1 to IgAN, little is known concerning IgA1-secreting subpopulations responsible for autoantigen production. The goal of this study is to identify IgA1-secreting and membrane subpopulations from single-cell transcriptomic analysis. We developed a novel single-cell analytics workflow to discern cells expressing IgA1 secreted isoform or membrane-bound isoform. Multiple approaches were compared to assess immunoglobulin-isotype identity in single cells, and multiple immunoglobulin heavy-chain genes expressed in the same cells were found. To better identify specific immunoglobulin heavy-chain transcripts, we merged a software platform called Alteryx with the existing single-cell R toolkit program Seurat. This process allowed for improved calls on IgA1-secreting subpopulations based on secreting versus membrane splice-variant expression levels. |
| Databáze: | Directory of Open Access Journals |
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