CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES >

2019 , Vol. 37 >Issue 1: 18 - 22

DOI: https://doi.org/10.12140/j.issn.1000-7423.2019.01.004

ORIGINAL ARTICLES

Leishmania infantum infection stimulates neutrophils to generate extracellular traps in mouse

  • Fu-rong WEI ,
  • Yue-tao YANG ,
  • Jun-yun WANG ,
  • Yan-juan WANG ,
  • Jia-ming PAN ,
  • Jian-ping CAO
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  • National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention; Chinese Center for Tropical Diseases Research; WHO Collaborating Centre for Tropical Diseases; National Centerfor International Research on Tropical Diseases, Ministry of Science and Technology; Key Laboratory of Parasite and Vector Biology, Ministry of Health, Shanghai 200025, China

Received date: 2018-10-26

  Online published: 2019-03-18

Supported by

Supported by the National Natural Science Foundation of China(No. 81772225) and the Fourth Round of Three?Year Public Health Action Plan of Shanghai, China(No. 15GWZK0101)

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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

Cite this article

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 . DOI: 10.12140/j.issn.1000-7423.2019.01.004

References

[1] Scott P.Leishmania--a parasitized parasite[J]. N Engl J Med, 2011, 364(18): 1773-1774.
[2] Faria MS, Reis FC, Lima AP.Toll-like receptors in Leishmania infections: guardians or promoters?[J]. J Parasitol Res, 2012, 2012: 930257.
[3] Carlsen ED, Liang Y, Shelite TR, et al. Permissive and protective roles for neutrophils in leishmaniasis[J]. Clin Exp Immunol, 2015, 182(2): 109-118.
[4] Hurrell BP, Regli IB, Tacchini-Cottier F.Different Leishmania species drive distinct neutrophil functions[J]. Trends Parasitol, 2016, 32(5): 392-401.
[5] Pompeu ML, Freitas LA, Santos ML, et al. Granulocytes in the inflammatory process of BALB/c mice infected by Leishmania amazonensis: a quantitative approach[J]. Acta Trop, 1991, 48(3): 185-193.
[6] Ribeiro-Gomes FL, Sacks D.The influence of early neutrophil-Leishmania interactions on the host immune response to infection[J]. Front Cell Infect Microbiol, 2012, 2: 59.
[7] Matte C, Olivier M.Leishmania-induced cellular recruitment during the early inflammatory response: modulation of proinflammatory mediators[J]. J Infect Dis, 2002, 185(5): 673-681.
[8] Thalhofer CJ, Chen Y, Sudan B, et al. Leukocytes infiltrate the skin and draining lymph nodes in response to the protozoan Leishmania infantum chagasi[J]. Infect Immun, 2011, 79(1): 108-117.
[9] Hurrell BP, Schuster S, Grun E, et al. Rapid sequestration of Leishmania mexicana by neutrophils contributes to the development of chronic Lesion[J]. PLoS Pathog, 2015, 11(5): e1004929.
[10] Tecchio C, Micheletti A, Cassatella MA.Neutrophil-derived cytokines: facts beyond expression[J]. Front Immunol, 2014, 5(1): 508.
[11] Kambayashi T, Laufer TM.A typical MHC class Ⅱ-expressing antigen-presenting cells: can anything replace a dendritic cell?[J]. Nat Rev Immunol, 2014, 14(11): 719-730.
[12] Hock BD, Taylor KG, Cross NB, et al. Effect of activated human polymorphonuclear leucocytes on T lymphocyte proliferation and viability[J]. Immunology, 2012, 137(3): 249-258.
[13] Puga I, Cols M, Barra CM, et al. B cell-helper neutrophils stimulate the diversification and production of immunoglobulin in the marginal zone of the spleen[J]. Nat Immunol, 2016, 13(2): 170-180.
[14] Papayannopoulos V.Neutrophil extracellular traps in immunity and disease[J]. Nat Rev Immunol, 2018, 18(2): 134-147.
[15] Guimaraes-Costa AB, DeSouza-Vieira TS, Paletta-Silva R, et al. 3′-nucleotidase/nuclease activity allows Leishmania parasites to escape killing by neutrophil extracellular traps[J]. Infect Immun, 2014, 82(4): 1732-1740.
[16] Guimaraes-Costa AB, Nascimento MT, Froment GS, et al. Leishmania amazonensis promastigotes induce and are killed by neutrophil extracellular traps[J]. Proc Natl Acad Sci U S A, 2009, 106(16): 6748-6753.
[17] Gabriel C, McMaster WR, Girard D, et al. Leishmania donovani promastigotes evade the antimicrobial activity of neutrophil extracellular traps[J]. J Immunol, 2010, 185(7): 4319-4327.
[18] Brinkmann V, Reichard U, Goosmann C, et al. Neutrophil extracellular traps kill bacteria[J]. Science, 2004, 303(5663): 1532-1535.
[19] Wang JY, Gao CH, Yang YT, et al. An outbreak of the desert sub-type of zoonotic visceral leishmaniasis in Jiashi, Xinjiang Uygur Autonomous Region, People’s Republic of China[J]. Parasitol Int, 2010, 59(3): 331-337.
[20] Lee KH, Cavanaugh L, Leung H, et al. Quantification of NETs-associated markers by flow cytometry and serum assays in patients with thrombosis and sepsis[J]. Int J Lab Hematol, 2018, 40(4): 392-399.
[21] Yipp BG, Petri B, Salina D, et al. Infection-induced NETosis is a dynamic process involving neutrophil multitasking in vivo[J]. Nat Med, 2012, 18(9): 1386-1393.
[22] Mayadas TN, Cullere X, Lowell CA.The multifaceted functions of neutrophils[J]. Annu Rev Pathol, 2014, 9(1): 181-218.
[23] Wong SL, Demers M, Martinod K, et al. Diabetes primes neutrophils to undergo NETosis, which impairs wound healing[J]. Nat Med, 2015, 21(7): 815-819.
[24] Garciaromo GS, Caielli S, Vega B, et al. Netting neutrophils are major inducers of type I IFN production in pediatric systemic lupus erythematosus [J]. Sci Transl Med, 2011, 3(73): 73ra20.
[25] 宋向楠, 刘彦虹. 中性粒细胞胞外诱捕网与相关疾病的研究进展[J]. 国际免疫学杂志, 2017, 40(1): 57-60.
[26] Gardinassi LG, DeSouza-Vieira TS, da Silva NO, et al. Molecular signatures of neutrophil extracellular traps in human visceral leishmaniasis[J]. Parasit Vectors, 2017, 10(1): 285.
[27] Morgado FN, Nascimento MT, Saraiva EM , et al. Are neutrophil extracellular traps playing a role in the parasite control in active american tegumentary leishmaniasis lesions?[J]. PLoS One, 2015, 10(7): e0133063.
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