The use of ErpY-like recombinant protein from Leptospira interrogans in the development of an immunodiagnostic test for swine leptospirosis

Padilha, Bárbara Couto Roloff; Simão, Henrique Queiroz; Oliveira, Thaís Larré; Hartwig, Daiane Drawanz

doi:10.1016/j.actatropica.2019.01.027

Cited by 9 publications

(13 citation statements)

References 24 publications

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“…To date, the role of ErpY-like protein in Leptospira pathogenesis has not been known; however, it is a potential vaccine and diagnostic candidate for leptospirosis in the specific host (18,19). We demonstrate that ErpY-like protein of Leptospira is a surface-exposed outer membrane lipoprotein, and its recombinant form of antigen can be implemented in serological diagnosis of leptospirosis in humans, bovines, and canines.…”

Section: Discussionmentioning

confidence: 92%

“…Such binding proteins benefit the bacteria in intervening thrombin-catalyzed clot formation or inhibiting complement activation, essential for successful establishment in the host and impeding the innate defense system. In a recent study, a protein annotated ErpY-like (LIC11966) in Leptospira was demonstrated to be an Fg-binding protein with diagnostic and subunit vaccine potential (17)(18)(19). The ErpY protein annotation originated from outer surface protein E/F-related proteins of another pathogenic spirochete, Borrelia burgdorferi (20).…”

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confidence: 99%

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Role of Supramolecule ErpY-Like Lipoprotein of Leptospira in Thrombin-Catalyzed Fibrin Clot Inhibition and Binding to Complement Factors H and I, and Its Diagnostic Potential

et al. 2019

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Leptospirosis is one of the most widespread zoonoses caused by pathogenic Leptospira spp. In this study, we report that the LIC11966/ErpY-like lipoprotein is a surface-exposed outer membrane protein exclusively present in pathogenic species of Leptospira. The recombinant ErpY (rErpY)-like protein is recognized by the immunoglobulins of confirmed leptospirosis sera of diverse hosts (human, bovine, and canine), suggesting the expression of the native leptospiral surface protein during infection. Circular dichroism of pure rErpY-like protein showed the secondary structural integrity to be uncompromised during the purification process. Analysis of the rErpY-like protein by native polyacrylamide gel electrophoresis, chemical cross-linking, dynamic light scattering, and field emission transmission electron microscopy demonstrated it undergoes supramolecular assembly. The rErpY-like protein can bind to diverse host extracellular matrices, and it presented a saturable and strong binding affinity (dissociation constant [KD] of 70.45 ± 4.13 nM) to fibrinogen, a central host plasma component involved in blood clotting. In the presence of the rErpY-like supramolecule, thrombin-catalyzed fibrin clot formation is inhibited up to 7%, implying its role in inhibiting blood coagulation during Leptospira infection. In addition, binding of the rErpY-like supramolecule to complement factors H and I suggests the protein also contributes to Leptospira evading innate host defense during infection by inactivating alternative complement pathways. This study reveals that rErpY-like protein is functionally active in the supramolecular state and performs moonlighting activity under the given in vitro conditions.

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Section: Discussionmentioning

confidence: 92%

mentioning

confidence: 99%

Role of Supramolecule ErpY-Like Lipoprotein of Leptospira in Thrombin-Catalyzed Fibrin Clot Inhibition and Binding to Complement Factors H and I, and Its Diagnostic Potential

et al. 2019

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“…The widely accepted minimum final dilution microscopic agglutination test (MAT) titre considered to be significant is 1:100, with titres ≥1:100 considered positive [ 11 , [20] , [21] , [22] ]. Using this titre as the cut-off, there was no serological evidence of active Leptospira infection or previous seroconversion in any of the 105 swine included in the present serological survey.…”

Section: Discussionmentioning

confidence: 99%

“…Swine are maintenance hosts for a range of Leptospira serovars including Canicola, Grippotyphosa, Icterohaemorrhagiae, and Pomona [ 11 ] while swine disease is associated with serovars Australis, Canicola, Grippotyphosa, Hardjo, Icterohaemorrhagiae, Pomona and Tarassovi [ 12 ]. Swine leptospirosis can be an endemic infection with little evidence of disease, but when first introduced to a naïve herd, or a herd with waning immunity, infertility, abortion, neonatal death and weakness can ensue, while leptospires can persist in the kidney and genital tract of carrier swine [ 12 ].…”

Section: Introductionmentioning

confidence: 99%

Absence of serological or molecular evidence of Leptospira infection in farmed swine in the Hong Kong Special Administrative Region

et al. 2021

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Leptospirosis is an important zoonotic disease with several maintenance host species including swine. A cross sectional survey was undertaken between January to October 2020 to investigate the prevalence of leptospirosis in farmed swine in the Hong Kong Special Administrative Region (HKSAR) of China. Serum samples were collected from swine on seven farms (15 swine per farm; ten multiparous sows and five twelve-week-old weaners), while kidney samples were collected from 64 swine submitted for routine post-mortem (26 farms; average 2.4 swine per farm, range 1–6). Microscopic agglutination tests (MAT) to a panel of 24 Leptospira antigens did not reveal any evidence of seroconversion at a titre of 1:100. Polymerase chain reaction (PCR) testing of the kidney samples for Leptospira DNA did not detect any evidence of infection. Bayesian methods were used to compute the probability that the leptospirosis prevalence in farmed swine in the HKSAR was <3%, given none of the 105 swine sampled were positive on the MAT. The results of this study demonstrate no serological or molecular evidence of leptospirosis in farmed swine in the HKSAR. Subsequent statistical analysis supports the conclusion that the prevalence of leptospirosis in farmed swine in the HKSAR is negligible at present.

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“…The development of diagnostic tests using recombinant proteins as ELISA antigens for leptospirosis in humans and other mammals was reported previously 24,25,36,37,54 . Most of the reported serological tests used proteins that are commonly found in pathogenic strains of Leptospira , with sensitivities ranging from 40% to 90% and specificity ranging from 75% to 100%.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of new leptospiral antigens for the diagnosis of equine leptospirosis: An approach using pan‐genomic analysis, reverse vaccinology and antigenic selection

Zilch

Lee

Bressan

et al. 2020

Equine Veterinary Journal

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Background The current gold standard diagnostic test for leptospirosis is the microscopic agglutination test (MAT), which has many drawbacks; therefore, the development of a better and easier serological test for leptospirosis is needed. Objectives To apply reverse vaccinology (RV) and antigenic selection on the assortment of leptospiral targets and evaluate their potential for use as reagents for the diagnosis of equine leptospirosis. Study design Cross‐sectional study. Methods The antigenic selection parameters were: proteins with antigenicity score ≥0.5 (VaxiJen), at least one B cell epitope and size between 10 and 275 KDa. New leptospiral proteins were cloned, expressed and serologically screened against equine sera (n = 128) on a single analysis and comparative combinations. Sensitivity (Se) and specificity (Sp), accuracy, positive predictive value (PPV) and negative predictive value (NPV) were calculated. A BLAST with nucleotide and protein sequences was used to identify the serovar or species specificity. Main limitations This cross‐sectional analysis had three main limitations: (a) The equine sera used in these tests were limited to sera submitted to the Animal Health Diagnosis Center and were only tested against seven serovars; (b) MAT results were considered being ‘perfect’, and the highest titre presented was considered being the infecting serovar, which may not hold true; (c) The strains used to represent the serovars and the limited number of different serovars and species included in the genetic analysis, which leads to the possibility that these proteins might be present in different species or serovars that perhaps would be seroprevalent in another geographic region. Conclusions The new leptospiral antigens described in this research could increase the sensitivity and specificity of ELISA for detection of Leptospira exposure and the detection of leptospirosis in horses along with support from other clinical signs. Some of these new antigens might be used to improve the detection of infecting serovar.

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The use of ErpY-like recombinant protein from Leptospira interrogans in the development of an immunodiagnostic test for swine leptospirosis

Cited by 9 publications

References 24 publications

Role of Supramolecule ErpY-Like Lipoprotein of Leptospira in Thrombin-Catalyzed Fibrin Clot Inhibition and Binding to Complement Factors H and I, and Its Diagnostic Potential

Role of Supramolecule ErpY-Like Lipoprotein of Leptospira in Thrombin-Catalyzed Fibrin Clot Inhibition and Binding to Complement Factors H and I, and Its Diagnostic Potential

Absence of serological or molecular evidence of Leptospira infection in farmed swine in the Hong Kong Special Administrative Region

Evaluation of new leptospiral antigens for the diagnosis of equine leptospirosis: An approach using pan‐genomic analysis, reverse vaccinology and antigenic selection

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