Leishmania infantum infection stimulates neutrophils to generate extracellular traps in mouse

doi:10.12140/j.issn.1000-7423.2019.01.004

Abstract

Abstract:

Objective To analyze the effect of Leishmania infantum infection on the differentiation of neutrophil extracellular traps (NETs) of mouse neutrophils under induction of MPA and the possible functions associated with the defense mechanism or pathogenesis of leishmaniasis. Methods Each of 30 BABL/c mice was intraperitoneally infected with 1 × 10⁸ L. infantum promastigotes in total volume of 0.5 ml. The peripheral blood were collected from the ether-anesthetized mice infected with L. infantum for 12 weeks or from normal control mice and the blood cells were stimulated with 100 nmol/L PMA for 3 h to generate NETs. The Red blood cells were lysed and the neutrophils were co-labeled with CD11b and Ly6G and NETs-generating neutrophils were labeled with H3Cit (citrullinated histone 3) and MPO (myeloperoxidase). The neutrophils and NET-generating neutrophils (H3Cit⁺MPO⁺) were quantitatively analyzed by flow cytometry. For fluorescent staining, the neutrophils were isolated from peripheral blood and the NET-generating neutrophils were induced with 100 nmol/L PMA for 3 h. The DNA, H3Cit and MPO in the neutrophils were stained with specific antibodies labeled with blue, red and green fluorescence, respectively and visualized under laser scanning confocal microscope. Results L. infantum infection significantly stimulated the differentiation of H3Cit⁺MPO⁺ NET neutrophils in the blood of mice after being stimulated with PMA (1.880 + 0.251)% compared to that in the normal control mice (0.623 + 0.087)% with statistical significance (P < 0.01). Laser confocal microscopic imaging analysis showed that the L. infantum infection stimulated more neutrophils of mouse to differentiate to NETs characterized by the filamentous DNA and H3Cit⁺MPO⁺ particles attached to the DNA skeleton structure, compared to neutrophils from normal mouse.

Key words: Leishmania infantum, Infection, Neutrophils, Neutrophil extracellular traps

CLC Number:

R382.22

Fu-rong WEI, Yue-tao YANG, Jun-yun WANG, Yan-juan WANG, Jia-ming PAN, Jian-ping CAO. Leishmania infantum infection stimulates neutrophils to generate extracellular traps in mouse[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2019, 37(1): 18-22.

Figures/Tables 2

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