Preliminary investigation of the clinical significance of neutrophils in patients with hepatic alveolar echinococcosis

doi:10.12140/j.issn.1000-7423.2026.01.006

Abstract

Abstract:

Objective To investigate neutrophil levels in peripheral blood and liver tissues among patients with hepatic alveolar echinococcosis (HAE), immune microenvironment characteristics, and their associations with the severity of HAE. Methods A total of 24 patients with AE who underwent surgical treatment in Qinghai University Affiliated Hospital from July 2024 to September 2025 (AE group) and 24 healthy controls (HC group) were included. All subjects in the AE group were definitively diagnosed by imaging and pathology. Subjects’ gender, age, neutrophil count, lymphocyte count, platelet count, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were collected, and the neutrophil to lymphocyte ratio (NLR) and systemic immune-inflammation index (SII) were calculated, with AE patients subjected to PMN classification. The correlations between peripheral blood clinical indicators were examined using the software GraphPad Prism 9 in the AE group. Peripheral blood, and surgically resected peri-lesional and distal liver tissues were sampled for hematoxylin and eosin (HE) staining, Masson staining, and immunohistochemical staining, and the proportions of areas stained positive for C-C motif chemokine ligand 2 (CCL2), C-X-C chemokine ligand 1 (CXCL1), and C-X-C motif chemokine receptor 2 (CXCR2) were estimated using the software QuPath-0.5.1, and the associations of CD16⁺ CD66b⁺ neutrophil levels in liver tissues with clinical indicators were examined using flow cytometry. The expression of inflammatory factors interleukin-8 (IL-8), IL-10, tumor necrosis factor alpha (TNF-α), granzyme B (GRZB), CCL2, elastase 2 (ELA2), intercellular adhesion molecule-1 (ICAM-1), gelatinase associated lipocalin (NGAL), matrix metalloproteinase (MMP-9) and myeloperoxidase (MPO) was measured peripheral blood and liver tissues using ELISA, and a correlation network analysis was performed. Results The peripheral blood lymphocyte [(1.60 ± 0.64) × 10⁹/L vs. (1.99 ± 0.62) × 10⁹/L; t = 2.036, P < 0.05] and platelet counts [(229.75 ± 62.20) × 10⁹/L vs. (260.29 ± 40.36) × 10⁹/L; t = 2.018, P < 0.05] were lower in the AE group than in the HC group, and the ALT [(35.54 ± 27.53) U/L vs. (24.17 ± 5.29) U/L; t = 2.319, P < 0.05], AST [(49.21 ± 47.63) U/L vs. (49.20 ± 47.64) U/L; t = 2.604, P < 0.05], NLR [2.06 (range, 1.48 to 3.27) vs. 1.67 ± 0.37; t = 2.317, P < 0.05] and SII [725.29 (range, 312.41 to 810.59) vs. 420.58 ± 44.81; t = 2.051, P < 0.05] were higher in the AE group than in the HC group. Correlation analysis revealed that NLR and SII strongly positively correlated with PMN classification (r = 0.67, P < 0.01), and weakly positively correlated with ALT (P < 0.05). Pathological examinations showed severe tissue structural disorders, hepatocyte coagulative necrosis, and fibrosis in peri-lesional liver tissues relative to distal tissues. Flow cytometry measured higher CD16⁺ CD66b⁺ neutrophil levels in peri-lesional liver tissues (20.49 ± 10.63) than in distal tissues (33.52 ± 11.23) (t = 6.923, P < 0.01), and the CD16⁺ CD66b⁺ neutrophil levels in peri-lesional liver tissues were positively correlated with PMN classification (r = 0.667, P < 0.05). Immunohistochemistry showed higher expression of CCL2 [(50.03 ± 15.30) vs. (40.99 ± 13.78); t = 6.492, P < 0.05], CXCL1 [(72.77 ± 13.45) vs. (56.94 ± 16.23); t = 5.527, P < 0.05], and CXCR2 [(74.49 ± 10.55) vs. (57.76 ± 15.26); t = 3.747, P < 0.05] in peri-lesional liver tissues than in distal tissues. ELISA measured elevated peripheral blood IL-6, IL-10, ELA2, NGAL, MMP-9, MPO, and TNF-α levels, high IL-8, IL-10, GRZB and CCL-2 expression and low NGAL and MMP-9 expression in peri-lesional liver tissues in the AE group. Correlation network analysis revealed strongly positive correlations between MPO and ICAM-1 (r = 0.98), and between MMP-9 and ELA2 (r = 0.88). Conclusion Remarkable infiltration of neutrophils driven by upregulation of chemokine axes (CXCL1, CXCR2, CCL2) is observed around the liver lesions among AE patients. Neutrophils drive cytokine storm and fibrosis remodeling around lesions through the synergistic effect of releasing oxidases and matrix-degrading enzymes. The pathological changes caused by the deterioration of local immune microenvironments determine the severity of AE (PNM classification) and synchronously project the significant increase of peripheral blood systemic inflammatory markers (NLR and SII).

Key words: Hepatic alveolar echinococcosis, Neutrophil, Inflammatory indicator, Chemokine, Immune microenvironment

CLC Number:

R532.32

FAN Yichen, ZHANG Chuanshan, HOU Jiao, TANG Zhaoyuan, LU Xuemei, HE Rongdong, JIAYIDAER Humaerhan, SONG Zhihao, KAN Mingxuan, WANG Mingjuan, SUN Li, WEN Hao. Preliminary investigation of the clinical significance of neutrophils in patients with hepatic alveolar echinococcosis[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2026, 44(1): 35-41.

Figures/Tables 5

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指标 Indicator	例数 No. cases	CD66b⁺ CD16⁺ /%	t/r值 t/r value	P值 P value
性别 Sex
男 Male	4	30.91 ± 15.35	t = 0.374	0.717
女 Female	7	31.92 ± 8.74
PNM			r = 0.667	0.004
P1N0M0	3	31.01 ± 5.95
P2N0M0	1	26.61 ± 0.00
P3N0M0	3	21.03 ± 4.60
P4N0M0	2	41.90 ± 7.12
P4N1M0	1	43.26 ± 0.00
P4N0M1	1	51.98 ± 0.00