Single-Cell Analysis Uncovers Abnormal Low-Density Neutrophil Differentiation in Myasthenia Gravis

Single-cell transcriptome profiling of peripheral blood immune cells has identified a significant expansion of low-density neutrophils and a concurrent reduction in CD4+ T cells among patients with myasthenia gravis, according to a study published in BMC Medical Genomics. In their analysis of an antibody-mediated autoimmune neuromuscular disorder where adaptive immune dysregulation is recognized but innate low-density neutrophil involvement has remained largely uncharacterized, the investigators evaluated blood samples from five anti-acetylcholine receptor antibody-positive patients and five age- and sex-matched healthy controls. Utilizing the BD Rhapsody platform for single-cell RNA sequencing alongside quality control procedures, the researchers classified twelve distinct immune cell populations across the cohort, documenting that low-density neutrophil numbers were significantly elevated whereas CD4+ T cells were significantly decreased (P < 0.01).

Differential gene expression analysis conducted with Presto revealed that low-density neutrophils exhibited distinctive transcriptomic programs enriched for neutrophil chemotaxis and neutrophil extracellular trap formation pathways. Upregulated genes specific to these low-density neutrophils included S100A8/A9, CSF3R, CD55, and CLEC12A. Subsequent subclustering focused specifically on the low-density neutrophil population resolved nine distinct cellular subsets, demonstrating clear shifts in subset representation between myasthenia gravis patients and healthy controls.

Within these nine subsets, pro-inflammatory CXCR2+ neutrophils and neutrophil extracellular trap-forming PADI4+ subsets, which exhibited elevated expression of trap-associated transcripts, were expanded in patient blood samples. In contrast, subsets displaying immunoregulatory features, specifically CD74+ and LTF+ subsets, were reduced in patients with myasthenia gravis. Pseudotime trajectory modeling performed using Monocle2 delineated a disease-specific maturation path characterized by a transition from CCND3+ cells through CXCR2+ intermediates into PADI4+ cells, indicating that low-density neutrophil differentiation is aberrantly altered in this disorder.

Analysis of gene regulatory networks and cell-to-cell communication further characterized the altered immune microenvironment using SCENIC and CellChat analytical frameworks. The transcription factor ELF2 demonstrated heightened regulatory activity, directing a network of 342 target genes, with 129 target genes displaying pseudotime-dependent expression tied to cell division and protein phosphorylation processes. Furthermore, intercellular signaling analyses revealed weakened ICAM1-mediated communication coupled with enhanced signaling along the ADGRE5–CD55/LPAR2 axis in patient low-density neutrophils, pointing to a shift toward pro-inflammatory and autoimmunity-promoting cellular interactions. Based on this ten-participant comparative dataset, the authors concluded that peripheral blood in myasthenia gravis is defined by low-density neutrophil subset imbalances, ELF2-driven aberrant differentiation, and dysregulated signaling networks that provide novel cellular markers and candidate therapeutic targets.

Subject of Research: Low-density neutrophil dynamics and immune signaling in myasthenia gravis

Article Title: Single-cell transcriptome analysis reveals aberrant low-density neutrophil differentiation and immune signaling dysregulation in myasthenia gravis

Article References: Yong, Y., Maimaiti, Z., Hao, M., Pei, F., Ma, N., Ma, Y., Du, Y., Habudule, Z., Liu, S., & Bi, F. (2026). Single-cell transcriptome analysis reveals aberrant low-density neutrophil differentiation and immune signaling dysregulation in myasthenia gravis. BMC Medical Genomics. https://doi.org/10.1186/s12920-026-02465-9

Image Credits: AI Generated

DOI: 10.1186/s12920-026-02465-9

Keywords: myasthenia gravis, low-density neutrophils, single-cell RNA sequencing, autoimmune disorders, CXCR2, PADI4, ELF2, cellular immunology, Single-cell, transcriptome, analysis, reveals