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1.
Vet Parasitol ; 327: 110136, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38290194

RESUMEN

Tick saliva has a pivotal function in parasitism. It has pharmacological and immunomodulatory properties, with several proteins reported in its composition. Thyroglobulin type-1 domain protease inhibitor (thyropin)-like proteins are found in tick saliva, but their function, properties and structures are poorly characterized. It has been reported that thyropins are capable of inhibiting cysteine peptidases present in antigen-presenting cells. To elucidate the role of thyropin-like proteins in ticks, we conducted in silico analysis and cloned an open reading frame from a thyropin-like protein found in Rhipicephalus microplus. The recombinant protein was successfully expressed, followed by immunological characterization and a vaccine trial against Rhipicephalus sanguineus in rabbits. Several differences are observed between thyropin-like proteins from hard and soft ticks, especially the number of thyroglobulin domains and predicted glycosylation pattern. Thyropin-like proteins also differ between postriata and metastriata ticks, the latter having a coil-domain at the C-terminal region and high number of predicted glycosylation sites. Overall, the data suggested divergence in thyropin-like proteins functions among ticks. The recombinant thyropin-like protein is immunogenic and the antibodies against it are able to recognize the native protein in tick saliva and tissues. While the recombinant protein does not elicit a protective response against R. sanguineus infestation, its characterization paves the way for further investigations aimed at determining the precise function of this protein in tick physiology.


Asunto(s)
Rhipicephalus sanguineus , Rhipicephalus , Infestaciones por Garrapatas , Animales , Conejos , Proteínas Recombinantes , Rhipicephalus/genética , Saliva/metabolismo , Tiroglobulina , Infestaciones por Garrapatas/prevención & control , Infestaciones por Garrapatas/veterinaria , Ensayos Clínicos Veterinarios como Asunto
2.
J Invest Dermatol ; 2023 Nov 22.
Artículo en Inglés | MEDLINE | ID: mdl-37996063

RESUMEN

The skin is the first host tissue that the tick mouthparts, tick saliva, and a tick-borne pathogen contact during feeding. Tick salivary glands have evolved a complex and sophisticated pharmacological arsenal, consisting of bioactive molecules, to assist blood feeding and pathogen transmission. In this work, persulcatin, a multifunctional molecule that targets keratinocyte function and hemostasis, was identified from Ixodes persulcatus female ticks. The recombinant persulcatin was expressed and purified and is a 25-kDa acidic protein with 2 Kunitz-type domains. Persulcatin is a classical tight-binding competitive inhibitor of proteases, targeting plasmin (Ki: 28 nM) and thrombin (Ki: 115 nM). It blocks plasmin generation on keratinocytes and inhibits their migration and matrix protein degradation; downregulates matrix metalloproteinase 2 and matrix metalloproteinase 9; and causes a delay in blood coagulation, endothelial cell activation, and thrombin-induced fibrinocoagulation. It interacts with exosite I of thrombin and reduces thrombin-induced endothelial cell permeability by inhibiting vascular endothelial-cadherin disruption. The multifaceted roles of persulcatin as an inhibitor and modulator within the plasminogen-plasmin system and thrombin not only unveil further insights into the intricate mechanisms governing wound healing but also provide a fresh perspective on the intricate interactions between ticks and their host organisms.

3.
Pathogens ; 12(9)2023 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-37764925

RESUMEN

Studies evaluating candidate tick-derived proteins as anti-tick vaccines in natural hosts have been limited due to high costs. To overcome this problem, animal models are used in immunization tests. The aim of this article was to review the use of rabbits as an experimental model for the evaluation of tick-derived proteins as vaccines. A total of 57 tick proteins were tested for their immunogenic potential using rabbits as models for vaccination. The most commonly used rabbit breeds were New Zealand (73.8%), Japanese white (19%), Californians (4.8%) and Flemish lop-eared (2.4%) rabbits. Anti-tick vaccines efficacy resulted in up to 99.9%. Haemaphysalis longicornis (17.9%) and Ornithodoros moubata (12.8%) were the most common tick models in vaccination trials. Experiments with rabbits have revealed that some proteins (CoAQP, OeAQP, OeAQP1, Bm86, GST-Hl, 64TRP, serpins and voraxin) can induce immune responses against various tick species. In addition, in some cases it was possible to determine that the vaccine efficacy in rabbits was similar to that of experiments performed on natural hosts (e.g., Bm86, IrFER2, RmFER2, serpins and serine protease inhibitor). In conclusion, results showed that prior to performing anti-tick vaccination trials using natural hosts, rabbits can be used as suitable experimental models for these studies.

4.
Animals (Basel) ; 13(12)2023 Jun 19.
Artículo en Inglés | MEDLINE | ID: mdl-37370541

RESUMEN

Recent advancements in molecular biology, particularly regarding massively parallel sequencing technologies, have enabled scientists to gain more insight into the physiology of ticks. While there has been progress in identifying tick proteins and the pathways they are involved in, the specificities of tick-host interaction at the molecular level are not yet fully understood. Indeed, the development of effective commercial tick vaccines has been slower than expected. While omics studies have pointed to some potential vaccine immunogens, selecting suitable antigens for a multi-antigenic vaccine is very complex due to the participation of redundant molecules in biological pathways. The expansion of ticks and their pathogens into new territories and exposure to new hosts makes it necessary to evaluate vaccine efficacy in unusual and non-domestic host species. This situation makes ticks and tick-borne diseases an increasing threat to animal and human health globally, demanding an urgent availability of vaccines against multiple tick species and their pathogens. This review discusses the challenges and advancements in the search for universal tick vaccines, including promising new antigen candidates, and indicates future directions in this crucial research field.

5.
Ticks Tick Borne Dis ; 11(4): 101445, 2020 07.
Artículo en Inglés | MEDLINE | ID: mdl-32354639

RESUMEN

In search of ways to address the increasing incidence of global acaricide resistance, tick control through vaccination is regarded as a sustainable alternative approach. Recently, a novel cocktail antigen tick-vaccine was developed based on the recombinant glutathione S-transferase (rGST) anti-sera cross-reaction to glutathione S-transferases of Rhipicephalus appendiculatus (GST-Ra), Amblyomma variegatum (GST-Av), Haemaphysalis longicornis (GST-Hl), Rhipicephalus decoloratus (GST-Rd) and Rhipicephalus microplus (GST-Rm). Therefore, the current study aimed to predict the shared B-cell epitopes within the GST sequences of these tick species. Prediction of B-cell epitopes and proteasomal cleavage sites were performed using immunoinformatics algorithms. The conserved epitopes predicted within the sequences were mapped on the homodimers of the respective tick GSTs, and the corresponding peptides were independently used for rabbit immunization experiments. Based on the dot blot assay, the immunogenicity of the peptides and their potential to be recognized by corresponding rGST anti-sera raised by rabbit immunization in a previous work were investigated. This study revealed that the predicted conserved B-cell epitopes within the five tick GST sequences were localized on the surface of the respective GST homodimers. The epitopes of GST-Ra, GST-Rd, GST-Av, and GST-Hl were also shown to contain a seven residue-long peptide sequence with no proteasomal cleavage sites, whereas proteasomal digestion of GST-Rm was predicted to yield a 4-residue fragment. Given that a few proteasomal cleavage sites were found within the conserved epitope sequences of the four GSTs, the sequences could also contain a T-cell epitope. Finally, the peptide and rGST anti-sera reacted against the corresponding peptide, confirming their immunogenicity. These data support the claim that the rGSTs, used in the previous study, contain conserved B-cell epitopes, which elucidates why the rGST anti-sera cross-reacted to non-homologous tick GSTs. Taken together, the data suggest that the B-cell epitopes predicted in this study could be useful for constituting epitope-based GST tick vaccines.


Asunto(s)
Epítopos de Linfocito B/inmunología , Glutatión Transferasa/inmunología , Inmunogenicidad Vacunal/inmunología , Ixodidae/inmunología , Control de Ácaros y Garrapatas , Vacunación/métodos , Secuencia de Aminoácidos , Animales , Epítopos de Linfocito B/metabolismo , Glutatión Transferasa/metabolismo , Ixodidae/enzimología , Complejo de la Endopetidasa Proteasomal
6.
Ticks Tick Borne Dis ; 11(3): 101378, 2020 05.
Artículo en Inglés | MEDLINE | ID: mdl-31982372

RESUMEN

Rhipicephalus appendiculatus, the brown ear tick, is an important disease vector of livestock in eastern, central and southern Africa. Rhipicephalus appendiculatus acaricide resistance requires the search for alternative methods for its control. Cystatins constitute a superfamily of cysteine peptidase inhibitors vital for tick blood feeding and development. These inhibitors were proposed as antigens in anti-tick vaccines. In this work, we applied structural and biochemical approaches to characterize a new cystatin named R. appendiculatus cystatin 2a (Racys2a). Structural modeling showed that this new protein possesses characteristic type 2 cystatin motifs, besides conservation of other structural patterns along the protein. Peptidase inhibitory assays with recombinant Racys2a showed modulation of tick and host cathepsins involved in blood digestion and immune system responses, respectively. A heterologous tick challenge with R. appendiculatus in rabbits immunized with recombinant Rhipicephalus microplus cystatin 2c (rBmcys2c) was performed to determine cross-reactivity. Histological staining showed that rBmcys2c vaccination caused damage to the gut, salivary gland and ovary tissues in R. appendiculatus. Furthermore, cystatin vaccine reduced the number of fully engorged adult females in 11.5 %. Consequently, strategies to increase the protection rate are necessary, including the selection of two or more antigens to compose a vaccine cocktail.


Asunto(s)
Proteínas de Artrópodos/genética , Rhipicephalus/genética , Cistatinas Salivales/genética , Vacunas/genética , Secuencia de Aminoácidos , Animales , Proteínas de Artrópodos/química , Proteínas de Artrópodos/metabolismo , Femenino , Filogenia , Conejos , Rhipicephalus/metabolismo , Cistatinas Salivales/química , Cistatinas Salivales/metabolismo , Alineación de Secuencia , Vacunas/química , Vacunas/metabolismo
7.
Vaccine ; 37(14): 1918-1927, 2019 03 28.
Artículo en Inglés | MEDLINE | ID: mdl-30824358

RESUMEN

Cocktail vaccines are proposed as an attractive way to increase protection efficacy against specific tick species. Furthermore, such vaccines made with different tick antigens have the potential of cross-protecting against a broad range of tick species. However, there are still limitations to the selection of immunogen candidates. Acknowledging that glutathione S-transferases (GSTs) have been exploited as vaccines against ticks and other parasites, this study aimed to analyze a GST-cocktail vaccine as a potential broad-spectrum tick vaccine. To constitute the GST-cocktail vaccine, five tick species of economic importance for livestock industry were studied (Rhipicephalus appendiculatus, Rhipicephalus decoloratus, Rhipicephalus microplus, Amblyomma variegatum, and Haemaphysalis longicornis). Tick GST ORF sequences were cloned, and the recombinant GSTs were produced in Escherichia coli. rGSTs were purified and inoculated into rabbits, and the immunological response was characterized. The humoral response against rGST-Rd and rGST-Av showed a stronger cross-reactivity against heterologous rGSTs compared to rGST-Hl, rGST-Ra, and rGST-Rm. Therefore, rGST-Rd and rGST-Av were selected for constituting an experimental rGST-cocktail vaccine. Vaccination experiment in rabbits showed that rGST-cocktail caused 35% reduction in female numbers in a Rhipicephalus sanguineus infestation. This study brings forward an approach to selecting immunogens for cocktail vaccines, and the results highlight rGST-Rd and rGST-Av as potentially useful tools for the development of a broad-spectrum tick vaccine.


Asunto(s)
Glutatión Transferasa/inmunología , Infestaciones por Garrapatas/prevención & control , Garrapatas/enzimología , Garrapatas/inmunología , Vacunas/inmunología , Animales , Reacciones Cruzadas/inmunología , Glutatión Transferasa/química , Glutatión Transferasa/genética , Sistemas de Lectura Abierta , Conejos , Rhipicephalus sanguineus/enzimología , Rhipicephalus sanguineus/inmunología , Vacunas/administración & dosificación
8.
Sci Rep ; 7(1): 17554, 2017 12 14.
Artículo en Inglés | MEDLINE | ID: mdl-29242567

RESUMEN

The cattle tick Rhipicephalus microplus is a hematophagous ectoparasite that causes important economic losses in livestock. Different species of ticks harbor a symbiont bacterium of the genus Coxiella. It was showed that a Coxiella endosymbiont from R. microplus (CERM) is a vertically transmitted mutualist symbiont, comprising 98% of the 16S rRNA sequences in both eggs and larvae. Sequencing of the bacterial genome revealed genes for biosynthetic pathways for several vitamins and key metabolic cofactors that may provide a nutritional complement to the tick host. The CERM was abundant in ovary and Malpighian tubule of fully engorged female. Tetracycline treatment of either the tick or the vertebrate host reduced levels of bacteria in progeny in 74% for eggs and 90% for larvae without major impact neither on the reproductive fitness of the adult female or on embryo development. However, CERM proved to be essential for the tick to reach the adult life stage, as under antibiotic treatment no tick was able to progress beyond the metanymph stage. Data presented here suggest that interference in the symbiotic CERM-R. microplus relationship may be useful to the development of alternative control methods, highlighting the interdependence between ticks and their endosymbionts.


Asunto(s)
Coxiella/fisiología , Rhipicephalus/microbiología , Simbiosis , Animales , Coxiella/efectos de los fármacos , Coxiella/genética , Femenino , Genoma Bacteriano/genética , Larva/efectos de los fármacos , Larva/crecimiento & desarrollo , Larva/microbiología , Ninfa/efectos de los fármacos , Ninfa/crecimiento & desarrollo , Ninfa/microbiología , Óvulo/efectos de los fármacos , Óvulo/crecimiento & desarrollo , Óvulo/microbiología , Rhipicephalus/crecimiento & desarrollo , Simbiosis/efectos de los fármacos , Tetraciclina/farmacología
9.
Ci. Rural ; 46(7): 1240-1248, jul. 2016.
Artículo en Inglés | VETINDEX | ID: vti-22598

RESUMEN

Rhipicephalus (Boophilus) species are monoxenous ticks with seasonal distribution in tropical and subtropical regions. For many years, Rhipicephalus microplus was considered as a single species; however, further analysis split these ticks into two distinct species. Because R. microplus and R. australis share similar attributes, it is hard to discriminate these two species and explain the changes in the classification of these parasites over the past decades. The reappearance of R. australis is an outcome of new research, which has afforded to better characterize these probably cryptic species. Evidence based on morphological features, the lack of conspecificity, microsatellite markers, mitochondrial 12S and 16S ribosomal DNA, and mitochondrial genome supports the re-classification of R. microplus as different species. Therefore, populations of R. microplus from Australia, Cambodia, Philippines, Indonesia, New Caledonia, Borneo, New Guinea, Tahiti and parts of Southeast Asia were recently reinstated as R. australis . Moreover, a better knowledge on the speciation between these two species could pave the way to important advances in tick control strategies.(AU)


As espécies pertencentes ao gênero Rhipicephalus (Boophilus) são carrapatos monoxenos de distribuição sazonal em regiões tropicais e subtropicais. Por muito anos, duas espécies de carrapatos foram consideradas como Rhipicephalus microplus. Contudo, estudos recentes reclassificaram esse carrapato em duas espécies: R. microplus e R. australis . Em razão de diversas semelhanças entre R. microplus e R. australis, distinguir essas duas espécies torna-se uma tarefa árdua, o que explica as mudanças de classificação dessas espécies nas últimas décadas. O reaparecimento da espécie R. australis surge com novas pesquisas, resultado de uma melhor caracterização dessas prováveis espécies crípticas. Evidências baseadas em análises das características morfológicas, na ausência de co-especificidade, em marcadores de microssatélites, no DNA ribossomal mitocondrial 12S e 16S, assim como no genoma mitocondrial, suportam a re-classificação de R. microplus como duas espécies distintas. Nesse sentido, populações de R. microplus da Austrália, Camboja, Nova Caledônia, Bornéo, Filipinas, Nova Guiné, Indonésia e Taiti foram recentemente renomeadas como R. australis . Além disso, um melhor entendimento sobre a especiação e localização dessas duas espécies pode trazer avanços importantes para melhorar as estratégias de controle desses carrapatos.(AU)


Asunto(s)
Animales , Rhipicephalus/clasificación , Rhipicephalus/genética , Rhipicephalus/fisiología , Garrapatas/clasificación , Garrapatas/genética , Garrapatas/fisiología , Clasificación
10.
Ciênc. rural ; Ciênc. rural (Online);46(7): 1240-1248, July 2016.
Artículo en Inglés | LILACS | ID: lil-780872

RESUMEN

ABSTRACT: Rhipicephalus ( Boophilus) species are monoxenous ticks with seasonal distribution in tropical and subtropical regions. For many years, Rhipicephalus micropluswas considered as a single species; however, further analysis split these ticks into two distinct species. Because R. microplusand R. australisshare similar attributes, it is hard to discriminate these two species and explain the changes in the classification of these parasites over the past decades. The reappearance of R. australisis an outcome of new research, which has afforded to better characterize these probably cryptic species. Evidence based on morphological features, the lack of conspecificity, microsatellite markers, mitochondrial 12S and 16S ribosomal DNA, and mitochondrial genome supports the re-classification of R. microplusas different species. Therefore, populations of R. microplusfrom Australia, Cambodia, Philippines, Indonesia, New Caledonia, Borneo, New Guinea, Tahiti and parts of Southeast Asia were recently reinstated as R. australis. Moreover, a better knowledge on the speciation between these two species could pave the way to important advances in tick control strategies.


RESUMO: As espécies pertencentes ao gênero Rhipicephalus (Boophilus) são carrapatos monoxenos de distribuição sazonal em regiões tropicais e subtropicais. Por muito anos, duas espécies de carrapatos foram consideradas como Rhipicephalus microplus.Contudo, estudos recentes reclassificaram esse carrapato em duas espécies: R. micropluse R. australis. Em razão de diversas semelhanças entre R. micropluse R. australis,distinguir essas duas espécies torna-se uma tarefa árdua, o que explica as mudanças de classificação dessas espécies nas últimas décadas. O reaparecimento da espécie R. australissurge com novas pesquisas, resultado de uma melhor caracterização dessas prováveis espécies crípticas. Evidências baseadas em análises das características morfológicas, na ausência de co-especificidade, em marcadores de microssatélites, no DNA ribossomal mitocondrial 12S e 16S, assim como no genoma mitocondrial, suportam a re-classificação de R. micropluscomo duas espécies distintas. Nesse sentido, populações de R. microplusda Austrália, Camboja, Nova Caledônia, Bornéo, Filipinas, Nova Guiné, Indonésia e Taiti foram recentemente renomeadas como R. australis. Além disso, um melhor entendimento sobre a especiação e localização dessas duas espécies pode trazer avanços importantes para melhorar as estratégias de controle desses carrapatos.

11.
PLoS One ; 10(6): e0130008, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26091260

RESUMEN

RNA interference-mediated gene silencing was shown to be an efficient tool for validation of targets that may become anti-tick vaccine components. Here, we demonstrate the application of this approach in the validation of components of molecular signaling cascades, such as the Protein Kinase B (AKT)/Glycogen Synthase Kinase (GSK) axis during tick embryogenesis. It was shown that heptane and hypochlorite treatment of tick eggs can remove wax, affecting corium integrity and but not embryo development. Evidence of AKT and GSK dsRNA delivery into de-waxed eggs of via electroporation is provided. Primers designed to amplify part of the dsRNA delivered into the electroporated eggs dsRNA confirmed its entry in eggs. In addition, it was shown that electroporation is able to deliver the fluorescent stain, 4',6-diamidino-2-phenylindole (DAPI). To confirm gene silencing, a second set of primers was designed outside the dsRNA sequence of target gene. In this assay, the suppression of AKT and GSK transcripts (approximately 50% reduction in both genes) was demonstrated in 7-day-old eggs. Interestingly, silencing of GSK in 7-day-old eggs caused 25% reduction in hatching. Additionally, the effect of silencing AKT and GSK on embryo energy metabolism was evaluated. As expected, knockdown of AKT, which down regulates GSK, the suppressor of glycogen synthesis, decreased glycogen content in electroporated eggs. These data demonstrate that electroporation of de-waxed R. microplus eggs could be used for gene silencing in tick embryos, and improve the knowledge about arthropod embryogenesis.


Asunto(s)
Técnicas de Silenciamiento del Gen , ARN Bicatenario/genética , Rhipicephalus/genética , Animales , Proteínas de Artrópodos/genética , Proteínas de Artrópodos/metabolismo , Electroporación , Femenino , Expresión Génica , Glucógeno Sintasa Quinasas/genética , Glucógeno Sintasa Quinasas/metabolismo , Heptanos/química , Óvulo/química , Óvulo/fisiología , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Interferencia de ARN , Solventes/química , Técnicas de Cultivo de Tejidos , Ceras/química
12.
Parasit Vectors ; 8: 122, 2015 Feb 24.
Artículo en Inglés | MEDLINE | ID: mdl-25889092

RESUMEN

BACKGROUND: Cystatins are a group of cysteine protease inhibitors responsible for physiological proteolysis regulation and present in a wide range of organisms. Studies about this class of inhibitors in parasites have contributed to clarify their roles in important physiological processes, like blood digestion and modulation of host immune response during blood feeding. Thus, cystatins are a subject of research on the development of new parasite control methods. Additionally, the characterization of proteins shared by different parasite species represents a valuable strategy to find potential targets in multi-species control methods. However, cystatin functions in ticks remain undetermined, especially in Rhipicephalus microplus and Ixodes ovatus, two species that affect livestock and human health, respectively. METHODS: Here we report the inhibitory profile of two R. microplus (BrBmcys2b and BrBmcys2c) and one I. ovatus (JpIocys2a) cystatins to commercial cathepsins B, C, and L. The presence of native cystatins in R. microplus tissues was analyzed using sera against recombinant BrBmcys2b and BrBmcys2c. Also, a peptide from JpIocys2a was synthesized for rabbit immunization, and this serum was used to analyze the cross antigenicity between R. microplus and I. ovatus cystatins. RESULTS: Enzymatic inhibition profile of tick cystatins shows a distinct modulation for cathepsins related to tick blood digestion and evasion of host immune response. Furthermore, BrBmcys2b was detected in saliva and different tissues along tick stages, while BrBmcys2c was detected mainly in gut from partially engorged R. microplus females, demonstrating a distinct pattern of cystatin expression, secretion and traffic between tick tissues. Moreover, phylogenetic analysis suggests that JpIocys2a belongs to the group of tick gut secreted cystatins. Finally, cross-antigenicity assays revealed that antibodies against the JpIocys2a peptide recognize native and recombinant R. microplus cystatins. CONCLUSION: The presence of these proteins in different tissues and their ability to differentially inhibit cathepsins suggest distinct roles for JpIocys2a, BrBmcys2b, and BrBmcys2c in blood digestion, egg and larvae development, and modulation of host immune response in tick physiology. The cross-antigenicity between native and recombinant cystatins supports further experiments using JpIocys2a, BrBmcys2b, and BrBmcys2c as vaccine antigens.


Asunto(s)
Cistatinas/inmunología , Inmunización/veterinaria , Ixodes/inmunología , Rhipicephalus/inmunología , Secuencia de Aminoácidos , Animales , Sangre , Cricetinae , Reacciones Cruzadas , Digestión , Femenino , Interacciones Huésped-Parásitos , Humanos , Ixodes/genética , Ixodes/fisiología , Masculino , Datos de Secuencia Molecular , Filogenia , Conejos , Proteínas Recombinantes , Rhipicephalus/genética , Rhipicephalus/fisiología , Alineación de Secuencia
13.
Biochim Biophys Acta ; 1830(3): 2574-82, 2013 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-23274741

RESUMEN

BACKGROUND: Tick embryogenesis is a metabolically intensive process developed under tightly controlled conditions and whose components are poorly understood. METHODS: In order to characterize the role of AKT (protein kinase B) in glycogen metabolism and cell viability, glycogen determination, identification and cloning of an AKT from Rhipicephalus microplus were carried out, in parallel with experiments using RNA interference (RNAi) and chemical inhibition. RESULTS: A decrease in glycogen content was observed when AKT was chemically inhibited by 10-DEBC treatment, while GSK3 inhibition by alsterpaullone had an opposing effect. RmAKT ORF is 1584-bp long and encodes a polypeptide chain of 60.1 kDa. Phylogenetic and sequence analyses showed significant differences between vertebrate and tick AKTs. Either AKT or GSK3 knocked down cells showed a 70% reduction in target transcript levels, but decrease in AKT also reduced glycogen content, cell viability and altered cell membrane permeability. However, the GSK3 reduction promoted an increase in glycogen content. Additionally, either GSK3 inhibition or gene silencing had a protective effect on BME26 viability after exposure to ultraviolet radiation. R. microplus AKT and GSK3 were widely expressed during embryo development. Taken together, our data support an antagonistic role for AKT and GSK3, and strongly suggest that such a signaling axis is conserved in tick embryos, with AKT located upstream of GSK3. GENERAL SIGNIFICANCE: The AKT/GSK3 axis is conserved in tick in a way that integrates glycogen metabolism and cell survival, and exhibits phylogenic differences that could be important for the development of novel control methods.


Asunto(s)
Proteínas de Artrópodos/genética , Glucógeno Sintasa Quinasa 3/genética , Glucógeno/metabolismo , Glucogenólisis/genética , Proteínas Proto-Oncogénicas c-akt/genética , Rhipicephalus/genética , Animales , Proteínas de Artrópodos/antagonistas & inhibidores , Proteínas de Artrópodos/metabolismo , Benzazepinas/farmacología , Línea Celular , Permeabilidad de la Membrana Celular/efectos de la radiación , Supervivencia Celular/efectos de la radiación , Clonación Molecular , Embrión no Mamífero , Regulación de la Expresión Génica/efectos de la radiación , Glucógeno/genética , Glucógeno Sintasa Quinasa 3/antagonistas & inhibidores , Glucógeno Sintasa Quinasa 3/metabolismo , Glucogenólisis/efectos de la radiación , Indoles/farmacología , Sistemas de Lectura Abierta , Oxazinas/farmacología , Filogenia , Proteínas Proto-Oncogénicas c-akt/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-akt/metabolismo , ARN Interferente Pequeño/genética , Rhipicephalus/embriología , Rhipicephalus/metabolismo , Homología de Secuencia de Aminoácido , Transducción de Señal/efectos de la radiación , Especificidad de la Especie , Rayos Ultravioleta
14.
Exp Parasitol ; 127(1): 113-8, 2011 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-20619263

RESUMEN

Recombinant Glutathione S-transferase of Haemaphysalis longicornis (rGST-Hl) was expressed in Escherichia coli, purified by affinity chromatography and used in the immunization of cattle. Western blot analysis showed positive antibody response in cattle immunized with rGST-Hl. The tests also showed that immunized bovine sera recognize native Rhipicephalus microplus proteins in different tissue extracts. Furthermore, the vaccine potential of rGST-Hl was investigated against infestation of Hereford cattle by R. microplus. Vaccination of cattle with rGST-Hl conferred partial cross-protection immunity against R. microplus. Considering the effect on number of engorged ticks, egg laying capacity and egg fertility, the overall efficacy of vaccination was of 57%, as compared with control group.


Asunto(s)
Enfermedades de los Bovinos/prevención & control , Glutatión Transferasa/inmunología , Ixodidae/inmunología , Rhipicephalus/inmunología , Infestaciones por Garrapatas/veterinaria , Algoritmos , Animales , Formación de Anticuerpos , Especificidad de Anticuerpos , Western Blotting/veterinaria , Bovinos , Enfermedades de los Bovinos/inmunología , Enfermedades de los Bovinos/parasitología , Femenino , Glutatión Transferasa/genética , Sueros Inmunes/inmunología , Ixodidae/enzimología , Infestaciones por Garrapatas/inmunología , Infestaciones por Garrapatas/prevención & control , Vacunas Sintéticas/inmunología
15.
Rev Bras Parasitol Vet ; 18(1): 1-7, 2009.
Artículo en Inglés | MEDLINE | ID: mdl-19500453

RESUMEN

The tick Rhipicephalus (Boophilus) microplus (formerly Boophilus microplus) is the major ectoparasite affecting livestock in America, Asia, Africa, and Oceania. Conventional tick control is based on the use of acaricides but immunization of bovines with tick gut proteins induces only a partial protective immune response. Based on this information, distinct research groups have explored the possibility of protecting the animals by inducing an immune response against other tick proteins. However, the antigens so far described do not induce the necessary protection for suppressing the use of acaricides. Currently, several groups are engaged in identifying new tick proteins to be used as targets for the development of new vaccines. This approach focuses on the enhancement of the immunogenicity of antigens already tested by incorporating new adjuvants or formulations and by searching for new antigens. This paper reviews the work done by Brazilian researchers to develop a vaccine against this tick.


Asunto(s)
Rhipicephalus , Infestaciones por Garrapatas/prevención & control , Vacunas , Animales , Ácido Aspártico Endopeptidasas , Precursores Enzimáticos , Serina Endopeptidasas
16.
Vet Parasitol ; 164(2-4): 282-90, 2009 Oct 14.
Artículo en Inglés | MEDLINE | ID: mdl-19560273

RESUMEN

The ticks Rhipicephalus (Boophilus) microplus and Haemaphysalis longicornis are blood-sucking ectoparasites of bovines, causing serious damages to the livestock production. The main control method for these ticks is based on acaricides. However, the use of vaccines has been studied as a promising control strategy. Calreticulin (CRT) is a multifunctional, predominantly intracellular protein present in almost all cells of animals. The secretion of CRT during feeding might be linked to the modulation of the parasite-host interaction. In the present study, recombinant CRTs of R. microplus (rBmCRT) and H. longicornis (rHlCRT) were expressed in Escherichia coli and purified by ion exchange chromatography and used for the immunization of bovines and mouse. ELISA demonstrated that both rCRTs are recognized by the sera of immunized bovines. In silico, despite the difference in amino acid sequences, antigenic index analysis of HlCRT and BmCRT using the Jameson-Wolf algorithm indicated that both proteins were very similar in antigenicity index, although six different epitopes between the tick CRTs have been inferred. These data were corroborated by competitive ELISA analyses, which suggest the presence of different epitopes within the proteins. Western blot analyses showed that anti-rBmCRT and anti-rHlCRT bovine sera also recognized the native proteins in larvae extracts and, moreover, sera of bovines immunized with saliva and extract of salivary glands recognized both recombinant CRTs. Thus, mouse and bovine immune system recognized rCRTs, resulting in the production of antibodies with similar specificity for both recombinant proteins, although different epitopes could be distinguished between rBmCRT and rHlCRT.


Asunto(s)
Calreticulina/clasificación , Calreticulina/inmunología , Ixodes/fisiología , Animales , Calreticulina/genética , Bovinos , Clonación Molecular , Simulación por Computador , Regulación de la Expresión Génica , Ratones , Filogenia , Proteínas Recombinantes
17.
Rev. bras. parasitol. vet ; 18(1): 1-7, Mar. 2009.
Artículo en Inglés | LILACS | ID: lil-606757

RESUMEN

The tick Rhipicephalus (Boophilus) microplus (formerly Boophilus microplus) is the major ectoparasite affecting livestock in America, Asia, Africa, and Oceania. Conventional tick control is based on the use of acaricides but immunization of bovines with tick gut proteins induces only a partial protective immune response. Based on this information, distinct research groups have explored the possibility of protecting the animals by inducing an immune response against other tick proteins. However, the antigens so far described do not induce the necessary protection for suppressing the use of acaricides. Currently, several groups are engaged in identifying new tick proteins to be used as targets for the development of new vaccines. This approach focuses on the enhancement of the immunogenicity of antigens already tested by incorporating new adjuvants or formulations and by searching for new antigens. This paper reviews the work done by Brazilian researchers to develop a vaccine against this tick.


O carrapato Rhipicephalus (Boophilus) microplus (anteriormente Boophilus microplus) é o principal ectoparasita que afeta bovinos na América, Ásia, África e Oceania e o seu controle é tradicionalmente realizado através do uso de acaricidas. Experimentos de imunização com proteínas do carrapato mostram que a resposta imune desenvolvida pelos bovinos vacinados protege, em parte, os animais do parasitismo. Baseado nessas observações, vários grupos de pesquisa exploram a possibilidade de proteger os animais pela indução de uma resposta imune contra proteínas do carrapato. Entretanto, os antígenos já caracterizados não asseguram o grau de proteção necessário para suprimir o uso de acaricidas. Portanto, esses grupos de pesquisa estão engajados na tentativa de identificar novas proteínas que possam ser utilizadas para o desenvolvimento de novas vacinas, as quais possam induzir maior imunogenicidade de que os antígenos já testados, através do uso de novas formulações e/ou pela incorporação de adjuvantes. O presente artigo apresenta uma revisão da literatura sobre os resultados obtidos por pesquisadores brasileiros no desenvolvimento de vacinas contra o carrapato.


Asunto(s)
Animales , Rhipicephalus , Infestaciones por Garrapatas/prevención & control , Vacunas , Ácido Aspártico Endopeptidasas , Precursores Enzimáticos , Serina Endopeptidasas
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