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Single-Cell Sequencing Confirms Transcripts and V_HDJ_H Rearrangements of Immunoglobulin Genes in Human Podocytes.

ABSTRACT: Most glomerular diseases are associated with inflammation caused by deposited pathogenic immunoglobulins (Igs), which are believed to be produced by B cells. However, our previous study indicated that the human podocyte cell line can produce IgG. In this study, we aimed to confirm the transcripts and characterize the repertoires of Igs in primary podocytes at single cell level. First, single-cell RNA sequencing of cell suspensions from "normal" kidney cortexes by a 10xGenomics Chromium system detected Ig transcripts in 7/360 podocytes and Ig gene segments in 106/360 podocytes. Then, we combined nested PCR with Sanger sequencing to detect the transcripts and characterize the repertoires of Igs in 48 single podocytes and found that five classes of Ig heavy chains were amplified in podocytes. Four-hundred and twenty-nine V_HDJ_H rearrangement sequences were analyzed; podocyte-derived Igs exhibited classic V_HDJ_H rearrangements with nucleotide additions and somatic hypermutations, biased VH1 usage and restricted diversity. Moreover, compared with the podocytes from healthy control that usually expressed one class of Ig and one V_HDJ_H pattern, podocytes from patients expressed more classes of Ig, V_HDJ_H patterns and somatic hypermutations. These findings suggested that podocytes can express Igs in normal condition and increase diversity in pathological situations.

SUBMITTER: Deng Z

PROVIDER: S-EPMC8064494 | biostudies-literature |

REPOSITORIES: biostudies-literature

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