Expression and characterisation of a Sarcoptes scabiei protein tyrosine kinase as a potential antigen for scabies diagnosis

Shen, Nengxing; He, Ran; Liu, Yuqing; Xu, Jing; He, Manli; Ren, Yongjun; Gu, Xiaobin; Lai, Weimin; Xie, Yue; Peng, Xuerong; Yang, Guangyou

doi:10.1038/s41598-017-10326-w

Cited by 10 publications

(9 citation statements)

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“…The optimal conditions for the rSsCLP5-based indirect ELISA were determined to be 4 μg/ml of rSsCLP5 protein, a 1:120 serum dilution, and a 1:3000 dilution of secondary antibody. Specific IgG antibody was detected by rSsCLP5-based indirect ELISA as previously described [ 37 ]. The results showed that the specific IgG antibodies increased at one week after the first vaccination in the rSsCLP5 vaccine groups, and levels were significantly higher than the antibody levels of three control groups ( F (5,63) = 37.285, P < 0.0001) (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Next, heat-induced epitope retrieval was accomplished by immersing sections in 0.01 M sodium citrate buffer (pH 6.0) at 95 °C for 20 min. The sections were incubated in blocking buffer (5% bovine serum albumin in PBS) for 1 h at room temperature before incubation overnight at 4 °C with specific rabbit anti-rSsCLP5 antibodies (preparation of rSsCLP5 polyclonal antibody as described previously [ 37 ]) covering the sections (diluted 1:100 in PBS). After washing three times with PBS, the sections were incubated with fluorescein isothiocyanate (FITC) goat anti-rabbit IgG (H+L; diluted 1:100; EarthOx, LLC, San Francisco, CA, USA) in 0.1% Evans Blue for 1 h at 37 °C in the dark.…”

Section: Methodsmentioning

confidence: 99%

“…A checkerboard titration study was carried out to determine the optimal conditions for the rSsCLP5 and serum [ 39 ]. ELISA procedures were performed as previously described [ 37 ]. We used the rSsCLP5 indirect ELISA to detect antibody IgG levels in serum samples at the time of vaccination and post-challenge.…”

Section: Methodsmentioning

confidence: 99%

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A chitinase-like protein from Sarcoptes scabiei as a candidate anti-mite vaccine that contributes to immune protection in rabbits

et al. 2018

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BackgroundScabies is caused by Sarcoptes scabiei burrowing into the stratum corneum of the host’s skin and is detrimental to the health of humans and animals. Vaccines are an attractive alternative to replace the acaricides currently used in their control.MethodsIn the present study, the S. scabiei chitinase-like protein 5 (SsCLP5) was characterized and recombinant SsCLP5 (rSsCLP5) was evaluated as a candidate vaccine protein for anti-mite protection in rabbits. The expression, characterization and immunolocalization of SsCLP5 were examined. Vaccination experiments were performed on three test groups (n = 12 per group) immunized with purified rSsCLP5. Control groups (n = 12 per group) were immunized with PBS, QuilA saponin or empty vector protein. After challenge, the inflammatory reaction and skin lesions were graded and rSsCLP5 indirect ELISA was used to detect antibody IgG levels in serum samples at the time of vaccination and post-challenge.ResultsThe results showed that rSsCLP5 had high immunoreactivity and immunogenicity. In S. scabiei, SsCLP5 had a wide distribution in the chewing mouthpart, legs and exoskeleton, especially the outer layer of the exoskeleton. Vaccination with rSsCLP5 resulted in 74.3% (26/35) of rabbits showing no detectable lesions after challenge with S. scabiei.ConclusionsOur data demonstrate that rSsCLP5 is a promising candidate for a recombinant protein-based vaccine against S. scabiei. This study also provides a method for studying scabies vaccine using rabbit as an animal model and a basis for screening more effective candidate proteins.Electronic supplementary materialThe online version of this article (10.1186/s13071-018-3184-y) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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A chitinase-like protein from Sarcoptes scabiei as a candidate anti-mite vaccine that contributes to immune protection in rabbits

et al. 2018

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“…Ordinary scabies (OS) and CS are two forms of scabies. OS is usually caused by a few mites, which makes the diagnosis very difficult, while CS is easy to diagnose based on the large number of mites and obvious clinical symptoms [ 8 , 9 ]. The development of serodiagnosis has seen serological diagnostic kits using mite extracts becoming commercially available [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

Comparative analysis of the allergenic characteristics and serodiagnostic potential of recombinant chitinase-like protein-5 and -12 from Sarcoptes scabiei

et al. 2021

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Background Scabies is caused by burrowing of the mite Sarcoptes scabiei into the stratum corneum. Currently, diagnosis via routine skin scraping is very difficult, and information on the allergenic identification of S. scabiei remains limited. Methods We performed comparative analysis of the serological diagnostic potential of recombinant S. scabiei chitinase-like protein-5 (rSsCLP5) and recombinant S. scabiei chitinase-like protein-12 (rSsCLP12) by measuring the levels of serum-specific IgG and IgE antibodies (Abs) as diagnostic markers. In addition, the allergenic characteristics of rSsCLP5 and rSsCLP12 were evaluated using IgE-binding experiments and skin tests. Results The IgE Abs-based indirect enzyme-linked immunosorbent assay (ELISA) methods showed high sensitivity and specificity: the rSsCLP5-based assay had 93.5% sensitivity and 94.4% specificity; the rSsCLP12-based assay had 100% sensitivity and 98.1% specificity. The specific IgE Abs in infested mouse sera could bind rSsCLP5 and rSsCLP12. In skin tests, rabbits in the rSsCLP5 and rSsCLP12 groups and positive control (histamine) groups exhibited allergic reactions. Most test sites in the rSsCLP12 group had edema, bleeding spots, and even ulcers or scabs, but such allergy symptoms were rare in the rSsCLP5 group. Moreover, the allergic history rabbit group had more severe allergic reactions and lower levels of IgE Abs compared to the healthy rabbit group in the same protein group. Conclusions These findings validate the use of IgE Abs to rSsCLP5 and rSsCLP12 as potentially useful markers for diagnosing scabies. Moreover, both rSsCLP5 and rSsCLP12 have allergenic properties, and the potential allergen rSsCLP12 is a stronger allergen than rSsCLP5.

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“…Allergens are the main factors eliciting the hosts pro-inflammatory response, and most of the allergen genes are hydrolase genes, having crucial roles in mite evasion and survival in the host [20, 21]. Moreover, there are several allergen and hydrolase genes in mites which have been identified to be potential diagnostic antigens and vaccine candidates [22–25]. Intriguingly, analysis of C. texanus allergen and hydrolase genes may provide a valuable database for exploring new potential diagnostic, prevention and treatment methods.…”

Section: Introductionmentioning

confidence: 99%

Transcriptome-based analysis of putative allergens of Chorioptes texanus

Xie

et al. 2019

Parasites Vectors

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BackgroundMites of the genus Chorioptes are non-burrowing and cause mange in a wide range of domestic and wild animals including cattle, horses, sheep, goats, panda, moose, camelids, mydaus and alpacas. Molecular biology and host-parasite interactions of Chorioptes texanus are poorly understood, and only a few C. texanus genes and transcript sequences are available in public databases including the allergen genes.MethodsChorioptes texanus RNA was isolated from mites, and the transcriptome of C. texanus was analyzed using bioinformatics tools. Chorioptes texanus unigenes were compared with the allergen protein sequences from the mite allergen database website to predict the potential allergens. Chorioptes texanus putative allergen unigenes were compared with hydrolase genes by building a C. texanus hydrolase gene library with the best match of the homologous sequences. Three allergen genes were cloned and expressed, their recombinant proteins were purified and their allergenic activities were preliminarily investigated.ResultsTranscriptome sequencing (RNA-Seq) of C. texanus was analyzed and results demonstrated that 33,138 unigenes were assembled with an average length of 751 bp. A total of 15,130 unigenes were annotated and 5598 unigenes were enriched in 262 KEGG signaling pathways. We obtained 209 putative allergen genes and 34 putative allergen-hydrolase genes. Three recombinant allergen proteins were observed to induce different degrees of allergic reactions on rabbit skin.ConclusionsThe present transcriptome data provide a useful basis for understanding the host-parasite interaction and molecular biology of the C. texanus mite. The allergenic activities of recombinant Euroglyphus maynei 1-like (Eur m 1-like) protein, Dermatophagoides ptreronyssinus 1-like (Der p 1-like) protein and Dermatophagoides ptreronyssinus 7-like (Der p 7-like) protein were preliminarily investigated by intradermal skin test. Meanwhile, differences in eosinophil counts were observed in different injected sites of the skin. The identification of putative allergen genes and hydrolase genes offers opportunities for the development of new diagnostic, prevention and treatment methods.

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Expression and characterisation of a Sarcoptes scabiei protein tyrosine kinase as a potential antigen for scabies diagnosis

Cited by 10 publications

References 49 publications

A chitinase-like protein from Sarcoptes scabiei as a candidate anti-mite vaccine that contributes to immune protection in rabbits

A chitinase-like protein from Sarcoptes scabiei as a candidate anti-mite vaccine that contributes to immune protection in rabbits

Comparative analysis of the allergenic characteristics and serodiagnostic potential of recombinant chitinase-like protein-5 and -12 from Sarcoptes scabiei

Transcriptome-based analysis of putative allergens of Chorioptes texanus

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