Frontiers in pediatrics · 2026
Diagnostic value of combined flow cytometry-based detection of peripheral blood lymphocyte subsets and cytokines for pediatric infectious mononucleosis.
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Tong Kai, Liu Xingran, Dong Jingxiao, Wang Qi, Hu Jing, Ma Jing
Аннотация
BACKGROUND: Pediatric infectious mononucleosis (IM) caused by Epstein-Barr virus (EBV) poses diagnostic challenges due to overlapping clinical presentations with other febrile illnesses. This study aimed to characterize immune function alterations in children with EBV-associated IM and evaluate the diagnostic performance of peripheral blood lymphocyte subsets and cytokine profiles, individually and in combination, to support early diagnosis.
METHODS: Between June 2024 and January 2026, pediatric patients with EBV-associated IM (IM group) and healthy children serving as controls (HC group) were prospectively enrolled. Lymphocyte subsets were quantified by flow cytometry, and plasma cytokines were measured by cytometric bead array (CBA). Inter-group differences were analyzed statistically, and receiver operating characteristic (ROC) curve analysis was used to evaluate diagnostic performance of individual and combined biomarkers.
RESULTS: A total of 61 children with IM and 32 healthy controls were enrolled. The predominant clinical manifestations in the IM group included fever (88.9%), pharyngitis (92.1%), lymphadenopathy (85.7%), tonsillar hypertrophy (90.4%), and periorbital edema (33.3%). Compared with the HC group, patients with IM demonstrated significantly elevated white blood cell counts, absolute lymphocyte counts, platelet counts, hemoglobin levels, and hepatic transaminases (ALT, AST) along with lactate dehydrogenase (LDH) (all P < 0.05). Immunophenotyping showed absolute counts of total lymphocytes, CD3⁺ T cells, CD8⁺ T cells, NK cells and percentages of CD3 and CD8 were significantly higher (P < 0.05) in the IM group, whereas CD4%, CD4/CD8 ratio, CD19%, and NK% were significantly decreased (P < 0.05). Cytokine analysis demonstrated significant elevation of IL-6, IL-10, TNF-α, IFN-γ, IL-17, IFN-α, and IL-12p70 in the IM group compared to the control group (P < 0.05). ROC analysis identified superior diagnostic performance for absolute counts of total lymphocytes (AUC=0.959), CD3⁺ T cells (AUC=0.957), and CD8⁺ T cells (AUC=0.972) among individual parameters. Combined lymphocyte subset analysis yielded an AUC of 0.995 (sensitivity 0.969, specificity 0.983), while combined cytokine assessment achieved an AUC of 0.980 (sensitivity 0.938, specificity 0.931), both substantially outperforming single biomarkers.
CONCLUSIONS: Pediatric IM is characterized by profound immune activation with marked CD8⁺ T cell expansion and elevated pro-inflammatory cytokines alongside the immunoregulatory cytokine IL-10. Combined assessment of lymphocyte subsets and cytokines significantly enhances IM diagnostic accuracy and holds substantial clinical utility.
METHODS: Between June 2024 and January 2026, pediatric patients with EBV-associated IM (IM group) and healthy children serving as controls (HC group) were prospectively enrolled. Lymphocyte subsets were quantified by flow cytometry, and plasma cytokines were measured by cytometric bead array (CBA). Inter-group differences were analyzed statistically, and receiver operating characteristic (ROC) curve analysis was used to evaluate diagnostic performance of individual and combined biomarkers.
RESULTS: A total of 61 children with IM and 32 healthy controls were enrolled. The predominant clinical manifestations in the IM group included fever (88.9%), pharyngitis (92.1%), lymphadenopathy (85.7%), tonsillar hypertrophy (90.4%), and periorbital edema (33.3%). Compared with the HC group, patients with IM demonstrated significantly elevated white blood cell counts, absolute lymphocyte counts, platelet counts, hemoglobin levels, and hepatic transaminases (ALT, AST) along with lactate dehydrogenase (LDH) (all P < 0.05). Immunophenotyping showed absolute counts of total lymphocytes, CD3⁺ T cells, CD8⁺ T cells, NK cells and percentages of CD3 and CD8 were significantly higher (P < 0.05) in the IM group, whereas CD4%, CD4/CD8 ratio, CD19%, and NK% were significantly decreased (P < 0.05). Cytokine analysis demonstrated significant elevation of IL-6, IL-10, TNF-α, IFN-γ, IL-17, IFN-α, and IL-12p70 in the IM group compared to the control group (P < 0.05). ROC analysis identified superior diagnostic performance for absolute counts of total lymphocytes (AUC=0.959), CD3⁺ T cells (AUC=0.957), and CD8⁺ T cells (AUC=0.972) among individual parameters. Combined lymphocyte subset analysis yielded an AUC of 0.995 (sensitivity 0.969, specificity 0.983), while combined cytokine assessment achieved an AUC of 0.980 (sensitivity 0.938, specificity 0.931), both substantially outperforming single biomarkers.
CONCLUSIONS: Pediatric IM is characterized by profound immune activation with marked CD8⁺ T cell expansion and elevated pro-inflammatory cytokines alongside the immunoregulatory cytokine IL-10. Combined assessment of lymphocyte subsets and cytokines significantly enhances IM diagnostic accuracy and holds substantial clinical utility.