Fig. 2: PBMCs scRNA-seq revealed elevated platelet count in MG and validated by peripheral blood laboratory test.

a Overview of the scRNA-seq experiment: PBMCs were isolated from three HC and three nMG patients, plus samples from two of the aforementioned MG patients when they achieved MMS post-treatment. b UMAP representation of scRNA-seq data illustrating the 16 cell types, including CD8+ T cells (CD8 CTL), CD4+ effector T cells (CD4 Tm), natural killer cells (NK), naïve B cells (Naïve B), CD8+ effector memory T cells (CD8 Tem), naïve CD8+ T cells (CD8 Naïve), gamma delta T Cells (γδ T), naïve CD4+ T cells (CD4 Naïve), CD14+ monocytes (CD14 MC), memory B Cells (Memory B), platelets, memory CD4+ T cells (Memory CD4 T), conventional dendritic cells (cDCs), plasmacytoid dendritic cells (pDCs), CD16+ monocytes (CD16 MC), and Plasma cells. c Stacked bar plot showing the relative abundance of PBMC subsets across nine samples. Each bar represents the proportion of identified cell clusters. d UMAP visualization illustrating the cluster abundance in PBMCs across three groups. The platelet cluster (encircled) is prominently enlarged in the MG group. e Boxplot analysis of platelet percentage by group, with statistical significance assessed via one-way ANOVA (two-sided, p = 0.12). The boxplot shows the minimum, first quartile, median, third quartile, and maximum values. Dashed lines indicate the comparative percentage of platelets in the same patient pre- and post-treatment. f A heatmap showcases the expression of genes related to platelet degranulation across the sample set. g Across a larger MG patient cohort, peripheral blood tests confirmed that nGMG patients (n = 65) exhibited higher platelet counts compared to HC (n = 25, p = 0.007) and MMS patients (n = 40, p = 0.0001). No significant differences were found between nOMG patients (n = 25) and the other groups. Data are presented as mean ± s.d., with individual biological replicates shown. P values were determined using ordinary one-way ANOVA followed by Tukey’s multiple comparisons test. Significance levels are denoted by asterisks: **p < 0.01; ****p < 0.0001. Non-significant results are not depicted. h The Circos plot presents the predicted cell-cell communication network, specifically mapping platelet-derived signaling to various immune cell populations in the MG (left, red), HC (middle, blue), and MMS (right, orange) groups. i, j Bubble plot visualization of platelet-mediated ligand-receptor interactions in MG. Each bubble represents an interaction between platelets and a specific immune cell type, with bubble color indicating the communication probability. The statistical inference method calculates communication probability by comparing the observed ligand-receptor expression against a null distribution generated through cell-label permutation, and assigns p values accordingly. i depicts incoming signals received by platelets, while j represents outgoing signals from platelets to other immune cells. The y-axis lists specific ligand-receptor pairs involved in these interactions, providing insight into platelet-driven immune modulation. PBMCs peripheral blood mononuclear cells, HC healthy controls, MMS minimal clinical status, nGMG immunotherapy-naïve generalized myasthenia gravis, nOMG immunotherapy-naïve ocular myasthenia gravis, IST immunological suppressive treatment, UMAP Uniform Manifold Approximation and Projection, Commun. Prob communication probability. Source data are provided as a Source data file.