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Neutrophils promote endothelial cell activation in pediatric Mycoplasma pneumoniae pneumonia
Xia Huang, Yifan Zhu, Yun Guo, Tian Lv, Haiyan Gu, Yingying Luo, Dan Li, Hang Sun, Deyu Zhao, Feng Liu
Xia Huang, Yifan Zhu, Yun Guo, Tian Lv, Haiyan Gu, Yingying Luo, Dan Li, Hang Sun, Deyu Zhao, Feng Liu
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Research Article Immunology Infectious disease

Neutrophils promote endothelial cell activation in pediatric Mycoplasma pneumoniae pneumonia

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Abstract

Mycoplasma pneumoniae pneumonia (MPP) can cause serious extrapulmonary complications, including life-threatening thrombosis. This study aimed to elucidate the roles of neutrophils and neutrophil extracellular traps (NETs) in vascular endothelial cell (EC) activation in pediatric MPP-associated thrombosis. We analyzed the relationship between neutrophils and thrombosis in children with MPP and used mouse models of neutrophilia (Csf3 plasmid injection), neutropenia (Csf3 deficient, Csf3–/–), and defective NETs formation (Pad4 deficient, Pad4–/–). The effects of neutrophils and NETs on EC activation were further examined in vivo, in vitro, and in human samples. Elevated neutrophil count was observed in patients with thrombosis and functioned as a potential diagnostic marker as well as a risk factor for MPP-associated thrombosis. EC activation was enhanced in MPP mice with neutrophilia but attenuated in neutropenic or Pad4–/– mice. NETs activated ECs through TLR2 and JAK/STAT3 signaling, and inhibition of NETs formation (Cl-amidine), TLR2 (C29), and JAK (upadacitinib) each attenuated this response. Strong correlations among neutrophils, NETs, EC activation, and thrombosis were observed in pediatric patients. These findings suggest that neutrophils promoted thrombosis in MPP via NETs-mediated EC activation involving TLR2 and JAK/STAT3 signaling. This study provides mechanistic insights into the inflammatory-thrombotic processes in MPP-associated thrombosis and offers a rationale for further investigation of neutrophils, NETs, TLR2, and JAK/STAT3 signaling in this context.

Authors

Xia Huang, Yifan Zhu, Yun Guo, Tian Lv, Haiyan Gu, Yingying Luo, Dan Li, Hang Sun, Deyu Zhao, Feng Liu

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Figure 1

Neutrophil count as a key indicator associated with thrombosis the Mycoplasma pneumoniae pneumonia (MPP) cohort.

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Neutrophil count as a key indicator associated with thrombosis the Mycop...
(A–D) Proportions of patients with bronchial obstruction (BO), necrosis, bronchiectasis, and mortality in the thrombosis versus nonthrombosis groups (n = 1,508). Categorical variables were compared using χ² tests and are presented as percentages. (E and F) Comparison of hospitalization duration (E) and costs (F) between thrombosis and nonthrombosis groups (n = 1,508). Data are presented as median (IQR) and compared using the Mann-Whitney U test. (G) LASSO coefficient profiles of candidate laboratory indicators. Each curve represents the trajectory of a variable’s regression coefficient as a function of log (λ) (n = 1508). The vertical dashed line indicates the optimal penalty parameter (λ) selected by cross-validation. (H) Forest plot of multivariable Firth’s penalized logistic regression analysis in the propensity score–matched (PSM) cohort (n = 303). Neutrophil count, C-reactive protein (CRP), D-dimer, and preadmission fever duration were independently associated with thrombosis. Odds ratios (ORs) with 95% CIs and corresponding P values are shown. (I) Comparison of peripheral neutrophil counts between patients with and without thrombosis. Violin plots illustrate the distribution of neutrophil levels in each group (n = 303). Statistical significance was assessed using the Mann-Whitney U test. (J) Receiver operating characteristic (ROC) curve evaluating the discriminative performance of neutrophil count for thrombosis (n = 303). Internal validation was performed using bootstrap resampling (1,000 iterations). The bootstrap-corrected area under the curve (AUC) was 0.899, with a 95% CI of 0.802–0.957. (K) Kaplan-Meier curves showing the cumulative probability of thrombosis stratified by neutrophil count (high versus low) in the PSM cohort (n = 303). Differences between groups were assessed using the log-rank test. The number of patients at risk at each time point is shown below the plot. ***P < 0.001, ****P < 0.0001.

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