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Derksen, N.

Publications and source records attributed to Derksen, N..

2 recordsLinked to original sources

Assessment of polymeric immunoglobulin A and M via detection of the joining chain

Polymeric immunoglobulin M (IgM) and A (IgA) play key roles in systemic and mucosal immunity, yet quantitative assessment of their polymeric forms has been hampered by the lack of robust, high-throughput assays. Polymerization of both isotypes implies incorporation of the joining chain (J chain), making direct detection of integrated J chain an attractive surrogate marker. Here, we report the generation and characterization of a novel panel of monoclonal antibodies targeting human J chain. Binding analyses revealed distinct antibody clusters with differential preferences for IgA-J and/or IgM-J. We developed sensitive ELISAs that allow reliable quantification of J-chain- containing IgM and IgA in recombinant preparations and complex biological samples such as serum and saliva. For IgM, assay performance in serum required mild dissociation of the IgM- CD5L complex, enabling accurate detection of integrated J chain. For IgA, clone 9G10 showed remarkable specificity for IgA-J, with minimal cross-reactivity to IgM. Application of these assays demonstrates that on average, 10% of circulating IgA is J-chain-containing, with proportional contributions of IgA1 and IgA2, and enables high-throughput measurement of antigen-specific polymeric IgA responses, exemplified by SARS-CoV-2 vaccination. These tools provide a long-needed platform to study polymeric antibody dynamics in health, infection, vaccination, and B-cell-driven diseases.

immunology↗

Transcriptome-Proteome analysis of human naive and memory B cell subsets reveal isotype and subclass-specific phenotypes

Antibodies produced by B cells aid in recognition and clearance of pathogens and is the cornerstone of vaccination strategies. Humans produce nine different antibody isotypes and their effector functions differ according to the type of antigen and route of exposure. Phenotypic variation between isotype-switched B cell subsets is expected but not studied in detail. To obtain a molecular definition of isotype-defined cell identity, we performed proteomics and transcriptomics on isotype-defined populations of human naive and memory B cells (MBCs): CD27-IgM+IgD+, CD27+CD38lo/-IgM+IgD+, CD27+CD38lo/-IgM+IgD-, and IgA1, IgA2, IgG1, IgG2, IgG3, and IgG4 MBCs (CD27+CD38lo/-Ig+). Combined proteome and transcriptome analysis revealed that mRNA and protein expression profiles separate isotype-defined B cell subsets according to their differentiation status. mRNA and protein expression levels correlated reasonably well for many genes. IgG4-switched B cells were most distinct from naive B cells in terms of mRNA as well as protein expression profiles. Besides a distinct expression profile of cytokine and Fc receptors, we identified a high expression of IgE-coding mRNA in IgG4-switched B cells. SDR16C5 was identified as uniquely upregulated in IgG4-switched B cells. Taken together, this study highlights the distinct phenotypic profile of IgG4-switched B cells.

immunology↗