The association of double-negative B cells with atherosclerosis and its severity.

Abstract

BACKGROUND: Double negative B (DNB) cells are a subset of mature B cells which does't express IgD and CD27 on surface. It is significantly expanded in various inflammatory related diseases. However, roles of DNB cells in atherosclerosis still remain unclear. Our objective is to evaluate the association between DNB cells and atherosclerosis and its severity. METHODS: A total of 138 patients who visited the Second Hospital of Dalian Medical University from June 2024 to December 2024 due to chest tightness and pain were enrolled. According to the results of coronary angiography examination, patients were divided into coronary heart disease (CHD) group and non-CHD group. The proportion of DNB cells in peripheral blood was detected using flow cytometry. Logistic regression and receiver operator characteristic (ROC) curve were utilized to explore the predictive value of the proportion of DNB cells for CHD and its severity. Moreover, atherosclerotic mice models were further constructed to explore the correlation between the proportion of DNB cells and atherosclerosis. To further reveal the potential mechanism, DNB cells were sorted for transcriptome sequencing analysis. RESULTS: Comparing with the non-CHD group, patients in the CHD group had a higher proportion of DNB cells (13.76&#xa0;&#xb1;&#xa0;5.48vs.7.58&#xa0;&#xb1;&#xa0;2.84, P&#xa0;<&#xa0;0.001). Logistic regression model showed that the proportion of DNB cells was an independent risk factor for CHD (OR:1.455, 95%CI:1.224-1.730). ROC curve indicated that the proportion of DNB cells had favor predictive value for CHD, better than hsCRP (high sensitivity C-reactive protein) (0.859 vs. 0.699). Our above findings were further validated in animal models. Comparing with the control group, the proportion of DNB cells in peripheral blood and spleen of atherosclerosis mice was increased and positively correlated with plaque area, blood lipids and inflammatory cytokines levels. GO and KEGG enrichment analyses revealed significant enrichment of inflammation- and chemokine-related pathways, with IL-6 participating in the greatest number of inflammatory cascades. Notably, CXCR4, which was strongly correlated with IL-6, may modulate IL-6 secretion via the MAPK and NF-&#x3ba;B signaling axes. CONCLUSIONS: DNB cells correlate with atherosclerosis severity, potentially via CXCR4/IL6-mediated inflammation. Underlying mechanisms require further elucidation.