Open AccessAssigning immunoglobulin class from single-cell transcriptomes in IgA1-secreting versus membrane subpopulations
Author(s) -
Colin Reily,
Nuo Xu,
David K. Crossman
Publication year - 2021
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2020-0044
Subject(s) - isotype , antibody , autoantibody , immunology , biology , b cell , gene isoform , immunoglobulin class switching , immunoglobulin a , immunoglobulin g , computational biology , gene , genetics , monoclonal antibody
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.