VWA Proteins of Leptospira Interrogans Induce Hemorrhage in Leptospirosis by Competitive Inhibition of VWF/GPIb-mediated Platelet Aggregation

Overview

Journal EBioMedicine

Specialty Biomedical Engineering

Date 2018 Oct 20

PMID 30337247

Citations 14

Authors

Jia-Qi Fang

Muhammad Imran

Wei-Lin Hu

David M Ojcius

Yang Li

Yu-Mei Ge

Kai-Xuan Li

Xuai Lin

Jie Yan

Affiliations

Soon will be listed here.

Abstract

Backgroud: Leptospira interrogans is the major causative agent of leptospirosis, a worldwide zoonotic disease. Hemorrhage is a typical pathological feature of leptospirosis. Binding of von Willebrand factor (vWF) to platelet glycoprotein-Ibα (GPIbα) is a crucial step in initiation of platelet aggregation. The products of L. interrogans vwa-I and vwa-II genes contain vWF-A domains, but their ability to induce hemorrhage has not been determined.

Methods: Human (Hu)-platelet- and Hu-GPIbα-binding abilities of the recombinant proteins expressed by L. interrogans strain Lai vwa-I and vwa-II genes (rLep-vWA-I and rLep-vWA-II) were detected by flowcytometry, surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC). Hu-platelet aggregation and its signaling kinases and active components were detected by lumiaggregometry, Western analysis, spectrophotometry and confocal microscopy. Hu-GPIbα-binding sites in rLep-vWA-I and rLep-vWA-II were identified by SPR/ITC measurements.

Findings: Both rLep-vWA-I and rLep-vWA-II were able to bind to Hu-platelets and inhibit rHu-vWF/ristocetin-induced Hu-platelet aggregation, but Hu-GPIbα-IgG, rLep-vWA-I-IgG and rLep-vWA-II-IgG blocked this binding or inhibition. SPR and ITC revealed a tight interaction between Hu-GPIbα and rLep-vWA-I/rLep-vWA-II with K values of 3.87 × 10-8.65 × 10 M. Hu-GPIbα-binding of rL-vWA-I/rL-vWA-II neither activated the PI3K/AKT-ERK and PLC/PKC kinases nor affected the NO, cGMP, ADP, Ca and TXA levels in Hu-platelets. G13/R36/G47 in Lep-vWA-I and G76/Q126 in Lep-vWA-II were confirmed as the Hu-GPIbα-binding sites. Injection of rLep-vWA-I or rLep-vWA-II in mice resulted in diffuse pulmonary and focal renal hemorrhage but this hemorrhage was blocked by rLep-vWA-I-IgG or rLep-vWA-II-IgG.

Interpretation: The products of L. interrogans vwa-I and vwa-II genes induce hemorrhage by competitive inhibition of vWF-mediated Hu-platelet aggregation.

Citing Articles

Sepsis - it is all about the platelets.

Cox D Front Immunol. 2023; 14:1210219.

PMID: 37350961 PMC: 10282552. DOI: 10.3389/fimmu.2023.1210219.

TcpC Inhibits M1 but Promotes M2 Macrophage Polarization via Regulation of the MAPK/NF-κB and Akt/STAT6 Pathways in Urinary Tract Infection.

Fang J, Ou Q, Wu B, Li S, Wu M, Qiu J Cells. 2022; 11(17).

PMID: 36078080 PMC: 9454685. DOI: 10.3390/cells11172674.

PD-L1 Regulates Platelet Activation and Thrombosis Caspase-3/GSDME Pathway.

Li Y, Xin G, Li S, Dong Y, Zhu Y, Yu X Front Pharmacol. 2022; 13:921414.

PMID: 35784685 PMC: 9240427. DOI: 10.3389/fphar.2022.921414.

Pulmonary haemorrhage as the earliest sign of severe leptospirosis in hamster model challenged with Leptospira interrogans strain HP358.

Philip N, Priya S, Jumah Badawi A, Mohd Izhar M, Mohtarrudin N, Ibrahim T PLoS Negl Trop Dis. 2022; 16(5):e0010409.

PMID: 35584087 PMC: 9116642. DOI: 10.1371/journal.pntd.0010409.

Next Generation Sequencing for Diagnosis of Leptospirosis Combined With Multiple Organ Failure: A Case Report and Literature Review.

Lu J, Hu J, Yu S, Li L Front Med (Lausanne). 2022; 8:756592.

PMID: 35145972 PMC: 8821090. DOI: 10.3389/fmed.2021.756592.

References

Goris M, Boer K, Duarte T, Kliffen S, Hartskeerl R . Human leptospirosis trends, the Netherlands, 1925-2008. Emerg Infect Dis. 2013; 19(3):371-8. PMC: 3647640. DOI: 10.3201/eid1903.111260. View

Nally J, Fishbein M, Blanco D, Lovett M . Lethal infection of C3H/HeJ and C3H/SCID mice with an isolate of Leptospira interrogans serovar copenhageni. Infect Immun. 2005; 73(10):7014-7. PMC: 1230959. DOI: 10.1128/IAI.73.10.7014-7017.2005. View

Smith J, Young M, Wilson K, Craig S . Leptospirosis following a major flood in Central Queensland, Australia. Epidemiol Infect. 2012; 141(3):585-90. PMC: 9151845. DOI: 10.1017/S0950268812001021. View

Miyata S, Goto S, Federici A, Ware J, Ruggeri Z . Conformational changes in the A1 domain of von Willebrand factor modulating the interaction with platelet glycoprotein Ibalpha. J Biol Chem. 1996; 271(15):9046-53. DOI: 10.1074/jbc.271.15.9046. View

Walker D, Valbuena G, Olano J . Pathogenic mechanisms of diseases caused by Rickettsia. Ann N Y Acad Sci. 2003; 990:1-11. DOI: 10.1111/j.1749-6632.2003.tb07331.x. View

Hu W, Ge Y, Ojcius D, Sun D, Dong H, Yang X . p53 signalling controls cell cycle arrest and caspase-independent apoptosis in macrophages infected with pathogenic Leptospira species. Cell Microbiol. 2013; 15(10):1642-59. DOI: 10.1111/cmi.12141. View

Kassegne K, Hu W, Ojcius D, Sun D, Ge Y, Zhao J . Identification of collagenase as a critical virulence factor for invasiveness and transmission of pathogenic Leptospira species. J Infect Dis. 2013; 209(7):1105-15. DOI: 10.1093/infdis/jit659. View

Kantonen S, Henriksen N, Gilson M . Evaluation and Minimization of Uncertainty in ITC Binding Measurements: Heat Error, Concentration Error, Saturation, and Stoichiometry. Biochim Biophys Acta Gen Subj. 2016; 1861(2):485-498. PMC: 5195854. DOI: 10.1016/j.bbagen.2016.09.002. View

Dutra F, Alves L, Rodrigues D, Fernandez P, de Oliveira R, Golenbock D . Hemolysis-induced lethality involves inflammasome activation by heme. Proc Natl Acad Sci U S A. 2014; 111(39):E4110-8. PMC: 4191786. DOI: 10.1073/pnas.1405023111. View

10.

Nally J, Chantranuwat C, Wu X, Fishbein M, Pereira M, Da Silva J . Alveolar septal deposition of immunoglobulin and complement parallels pulmonary hemorrhage in a guinea pig model of severe pulmonary leptospirosis. Am J Pathol. 2004; 164(3):1115-27. PMC: 1614724. DOI: 10.1016/S0002-9440(10)63198-7. View

11.

Satoh K, Asazuma N, Yatomi Y, Fujimura Y, Miura S, Titani K . Activation of protein-tyrosine kinase pathways in human platelets stimulated with the A1 domain of von Willebrand factor. Platelets. 2000; 11(3):171-6. DOI: 10.1080/095371000403116. View

12.

Choy H, Kelley M, Chen T, Moller A, Matsunaga J, Haake D . Physiological osmotic induction of Leptospira interrogans adhesion: LigA and LigB bind extracellular matrix proteins and fibrinogen. Infect Immun. 2007; 75(5):2441-50. PMC: 1865782. DOI: 10.1128/IAI.01635-06. View

13.

Li Z, Delaney M, OBrien K, Du X . Signaling during platelet adhesion and activation. Arterioscler Thromb Vasc Biol. 2010; 30(12):2341-9. PMC: 3085271. DOI: 10.1161/ATVBAHA.110.207522. View

14.

Zhang L, Zhang C, Ojcius D, Sun D, Zhao J, Lin X . The mammalian cell entry (Mce) protein of pathogenic Leptospira species is responsible for RGD motif-dependent infection of cells and animals. Mol Microbiol. 2012; 83(5):1006-23. DOI: 10.1111/j.1365-2958.2012.07985.x. View

15.

Haake D, Levett P . Leptospirosis in humans. Curr Top Microbiol Immunol. 2014; 387:65-97. PMC: 4442676. DOI: 10.1007/978-3-662-45059-8_5. View

16.

Yin H, Stojanovic A, Hay N, Du X . The role of Akt in the signaling pathway of the glycoprotein Ib-IX induced platelet activation. Blood. 2007; 111(2):658-65. PMC: 2200862. DOI: 10.1182/blood-2007-04-085514. View

17.

Du P, Li S, Ojcius D, Li K, Hu W, Lin X . A novel Fas-binding outer membrane protein and lipopolysaccharide of Leptospira interrogans induce macrophage apoptosis through the Fas/FasL-caspase-8/-3 pathway. Emerg Microbes Infect. 2018; 7(1):135. PMC: 6066479. DOI: 10.1038/s41426-018-0135-9. View

18.

Mendonca R, Silveira A, Conran N . Red cell DAMPs and inflammation. Inflamm Res. 2016; 65(9):665-78. DOI: 10.1007/s00011-016-0955-9. View

19.

Brophy D, Martin E, Gehr T, Best A, Paul K, Carr Jr M . Thrombin generation time is a novel parameter for monitoring enoxaparin therapy in patients with end-stage renal disease. J Thromb Haemost. 2006; 4(2):372-6. DOI: 10.1111/j.1538-7836.2006.01731.x. View

20.

Li Z, Zhang G, Feil R, Han J, Du X . Sequential activation of p38 and ERK pathways by cGMP-dependent protein kinase leading to activation of the platelet integrin alphaIIb beta3. Blood. 2005; 107(3):965-72. PMC: 1464421. DOI: 10.1182/blood-2005-03-1308. View