Leptospira borgpetersenii hybrid leucine-rich repeat protein: Cloning and expression, immunogenic identification and molecular docking evaluation

Sritrakul, Tepyuda; Nitipan, Supachai; Wajjwalku, Worawidh; La-ard, Anchalee; Suphatpahirapol, Chattip; Petkarnjanapong, Wimol; Ongphiphadhanakul, Boonsong; Prapong, Siriwan

doi:10.1016/j.mimet.2017.09.005

Cited by 4 publications

(11 citation statements)

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“…Leucine-rich repeats of bacterial surface proteins also serve as common pattern recognition motifs of host cell receptors, as reported in human scavenger receptor gp340 by LrrG and E-cadherin by rLRR20 [ 34 , 60 ]. Both rKU_Sej_LRR_2012M and rhKU_Sej_LRR_2271 proteins contain the LPXAG motif, and the two leptospiral LRR proteins characterized in this study had been reported to exhibit rapid induction of specific humoral immune responses in immunized rabbits as well as in hamsters of this report [ 8 , 37 , 38 , 39 , 40 ]. Although this report did not investigate the virulence of rKU_Sej_LRR_2012M and rhKU_Sej_LRR_2271 proteins in pathogenic Leptospira infection, it has been reported that the LRR domain-containing protein family is vital for the virulence of pathogenic Leptospira species [ 34 , 35 ].…”

Section: Discussionmentioning

confidence: 89%

“…Two LRR proteins, the rKU_Sej_LRR_2012M and rhKU_Sej_LRR_2271 proteins, were expressed from pET161_hKU_R21M_2012 and pET160_hKU_R21_2271 plasmids in Escherichia coli BL21 Star TM (DE3) expression systems, respectively [ 8 , 38 , 39 ] with a minor modification. Both proteins were expressed as 6xHis-Lumio-TEV fusion proteins.…”

Section: Methodsmentioning

confidence: 99%

“…Both proteins were expressed as 6xHis-Lumio-TEV fusion proteins. The proteins were purified by immobilized metal ion affinity chromatography (IMAC) with Protino ® Ni-TED Resin (Macherey-Nagel, Dueren, Germany) as previously described [ 38 ]. The 6xHis-Lumio-TEV was cleaved with TEV protease (NEB#P8112) after the purification step as per the manufacturer’s protocol.…”

Section: Methodsmentioning

confidence: 99%

“…The immunogenic epitopes were identified by in silico studies from two leptospiral LRR proteins, the rKU_Sej_LRR_2012M and rKU_Sej_LRR_2271 proteins, from L. borgpetersenii serogroup Sejroe genome [ 37 , 38 , 39 ]. In addition, the two identified LRR proteins have cross-reactive immunity with rabbit hyperimmune sera against Leptospira serovars Icterohaemorrhagiae, Javanica, serogroup Sejroe, and serovars Bratislava, Icterohaemorrhagiae, and serogroup Sejroe, respectively [ 38 , 40 ].…”

Section: Introductionmentioning

confidence: 99%

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Leptospira borgpetersenii Leucine-Rich Repeat Proteins Provide Strong Protective Efficacy as Novel Leptospiral Vaccine Candidates

et al. 2022

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Leucine-rich repeat (LRR) proteins are advocated for being assessed in vaccine development. Leptospiral LRR proteins were identified recently in silico from the genome of Leptospira borgpetersenii serogroup Sejroe, the seroprevalence of leptospiral infections of cattle in Thailand. Two LRR recombinant proteins, rKU_Sej_LRR_2012M (2012) and rhKU_Sej_LRR_2271 (2271), containing predicted immunogenic epitopes, were investigated for their cross-protective efficacies in an acute leptospirosis model with heterologous Leptospira serovar Pomona, though, strains from serogroup Sejroe are host-adapted to bovine, leading to chronic disease. Since serovar Pomona is frequently reported as seropositive in cattle, buffaloes, pigs, and dogs in Thailand and causes acute and severe leptospirosis in cattle by incidental infection, the serogroup Sejroe LRR proteins were evaluated for their cross-protective immunity. The protective efficacies were 37.5%, 50.0%, and 75.0% based on the survival rate for the control, 2012, and 2271 groups, respectively. Sera from 2012-immunized hamsters showed weak bactericidal action compared to sera from 2271-immunized hamsters (p < 0.05). Therefore, bacterial tissue clearances, inflammatory responses, and humoral and cell-mediated immune (HMI and CMI) responses were evaluated only in 2271-immunized hamsters challenged with virulent L. interrogans serovar Pomona. The 2271 protein induced prompt humoral immune responses (p < 0.05) and leptospiral tissue clearance, reducing tissue inflammation in immunized hamsters. In addition, protein 2271 and its immunogenic peptides stimulated splenocyte lymphoproliferation and stimulated both HMI and CMI responses by activating Th1 and Th2 cytokine gene expression in vaccinated hamsters. Our data suggest that the immunogenic potential renders rhKU_Sej_LRR_2271 protein a promising candidate for the development of a novel cross-protective vaccine against animal leptospirosis.

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Section: Discussionmentioning

confidence: 89%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Leptospira borgpetersenii Leucine-Rich Repeat Proteins Provide Strong Protective Efficacy as Novel Leptospiral Vaccine Candidates

et al. 2022

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“…The solution was centrifuged at 10,000× g for 30 min at 20 °C to collect the dissolved proteins. Since the rKU_Sej_LRR_2012M protein was expressed as 6× His-Lumio-TEV fusion proteins, the dissolved proteins were purified by the immobilized metal ion affinity chromatography (IMAC) with Protino ® Ni-TED Resin (Macherey-Nagel, Düren, Germany), according to the manufacturer’s protocol with minor modifications, as previously described [ 45 ]. The 6× His-Lumio-TEV was cleaved with TEV protease (NEB#P8112, NEB, Ipswich, MA, USA).…”

Section: Methodsmentioning

confidence: 99%

Leptospira borgpetersenii Leucine-Rich Repeat Proteins and Derived Peptides in an Indirect ELISA Development for the Diagnosis of Canine Leptospiral Infections

Sripattanakul

Prapong²,

Kamlangdee³

et al. 2022

TropicalMed

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Domestic and stray dogs can be frequently infected by Leptospira, and thus may represent a source for transmission of this zoonotic disease in Thailand. Here, we have used peptides derived from a recombinant leucine-rich repeat (LRR) protein of Leptospira, rKU_Sej_LRR_2012M, for the development of an indirect enzyme-linked immunosorbent assay (ELISA) aimed at detecting antibodies against Leptospira interrogans, L. borgpetersenii and L. biflexa, the three major seroprevalences in Thai dogs. The rKU_Sej_LRR_2012M protein is recognized by hyperimmune sera against several leptospiral serovars. The epitope peptides of the rKU_Sej_LRR_2012M showed binding affinities with lower IC50 values than peptides of known antigenic protein LipL32. Four peptides, 2012-3T, 2012-4B, 2012-5B and pool 2012-B, were specifically recognized by rabbit hyperimmune sera against nine serovars from three Leptospira spp. The indirect peptide-based ELISAs with these four peptides were evaluated with the LipL32 ELISA by using a receiver–operator curve (ROC) analysis. All peptides had an area under the curve of ROC (AUC) greater than 0.8, and the sum of sensitivity and specificity for each peptide was greater than 1.5. The degree of agreement of 2012-3T and pool 2012-B and 2012-4B and 2012-5B peptides were in moderate-to-good levels with kappa values of 0.41–0.60 and 0.61–0.80, when compared with LipL32, respectively. This finding would suggest an excellent capability of the 2012-4B and 2012-5B peptide-based ELISAs assay for the diagnosis of canine leptospiral infections.

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New potent epitopes from Leptospira borgpetersenii for the stimulation of humoral and cell-mediated immune responses: Experimental and theoretical studies

Tansiri

Sritrakul

Saparpakorn

et al. 2021

Informatics in Medicine Unlocked

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Leptospira borgpetersenii hybrid leucine-rich repeat protein: Cloning and expression, immunogenic identification and molecular docking evaluation

Cited by 4 publications

References 30 publications

Leptospira borgpetersenii Leucine-Rich Repeat Proteins Provide Strong Protective Efficacy as Novel Leptospiral Vaccine Candidates

Leptospira borgpetersenii Leucine-Rich Repeat Proteins Provide Strong Protective Efficacy as Novel Leptospiral Vaccine Candidates

Leptospira borgpetersenii Leucine-Rich Repeat Proteins and Derived Peptides in an Indirect ELISA Development for the Diagnosis of Canine Leptospiral Infections

New potent epitopes from Leptospira borgpetersenii for the stimulation of humoral and cell-mediated immune responses: Experimental and theoretical studies

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