Differential protein expression in ovaries of uninfected and Babesia-infected southern cattle ticks, Rhipicephalus (Boophilus) microplus (original) (raw)
Related papers
Veterinary Parasitology, 2008
Differences in protein expression in midgut tissue of uninfected and Babesia bovis-infected southern cattle ticks, Rhipicephalus (Boophilus) microplus, were investigated in an effort to establish a proteome database containing proteins involved in successful pathogen transmission. The electrophoretic separation of midgut membrane proteins was greatly improved by using liquid-phase isoelectric focusing combined with one-dimensional or two-dimensional (2-D) gel electrophoresis. A selection of differentially expressed proteins were subjected to analysis by capillary-HPLC-electrospray tandem mass spectrometry (HPLC-ESI-MS/MS). Among the identified Babesia-affected tick midgut proteins were six proteins that are implicated in signaling processes, including three Ca 2+-binding proteins, a guanine nucleotide-binding protein, a protein with signal peptide activity and a translocon-associated receptor protein. Up-regulation of five metabolic enzymes indicated parasite-induced changes in electron and proton transport, protein processing and retinoic acid metabolism. Among the down-regulated proteins were a molecular chaperone, a cytoskeletal protein and a multifunctional protein of the prohibitin family. Identification of these proteins may provide new insights into the molecular interactions between B. bovis and its tick vector, and could lead to identification of anti-tick and transmission-blocking vaccine candidates.
Characterization of the tick-pathogen interface by quantitative proteomics
Ticks and Tick-borne Diseases, 2012
Ticks are vectors of pathogens that affect human and animal health worldwide. Ticks and the pathogens they transmit have co-evolved molecular interactions involving genetic traits of both the tick and the pathogen that mediate their development and survival. Proteomics and genomics studies of infected ticks are required to understand tick-pathogen interactions and identify potential vaccine antigens to control tick infestations and pathogen transmission. In this paper, the application of quantitative proteomics to characterize differential protein expression in ticks and cultured tick cells in response to pathogen infection is reviewed. Analyses using (a) two-dimensional differential in gel electrophoresis (DIGE) labeling and (b) protein one-step in gel digestion, peptide iTRAQ labeling, and isoelectric focusing fractionation, both followed by peptide and protein identifications by mass spectrometry resulted in the identification of host, pathogen, and tick proteins differentially expressed in response to infection. Although at its infancy, these results showed that quantitative proteomics is a powerful approach to characterize the tick-pathogen interface and demonstrated pathogen and tick-specific differences in protein expression in ticks and cultured tick cells in response to pathogen infection.
Proteomics Characterization of Tick-Host-Pathogen Interactions
Methods in Molecular Biology, 2014
Ticks are blood-feeding arthropod ectoparasites of wild and domestic animals that transmit disease-causing pathogens to humans and animals worldwide and a good model for the characterization of tick-host-pathogen interactions. Tick-host-pathogen interactions consist of dynamic processes involving genetic traits of hosts, pathogens, and ticks that mediate their development and survival. Proteomics provides information on the protein content of cells and tissues that may differ from results at the transcriptomics level and may be relevant for basic biological studies and vaccine antigen discovery. In this chapter, we describe various methods for protein extraction and for proteomics analysis in ticks based on one-dimensional gel electrophoresis to characterize tick-host-pathogen interactions. Particularly relevant for this characterization is the use of blood-fed ticks. Therefore, we put special emphasis on working with replete ticks collected after feeding on vertebrate hosts.
Journal of insect …, 2010
The continued development of effective anti-tick vaccines remains the most promising prospect for the control of the cattle tick, Rhipicephalus (Boophilus) microplus. A vaccine based on midgut proteins could interfere with successful tick feeding and additionally interfere with midgut developmental stages of Babesia parasites, providing opportunities for the control of both the tick and the pathogens it transmits. Midgut proteins from partially fed adult female cattle ticks were analysed using a combination of 2-DE and gel-free LC-MS/MS. Analysis of the urea-soluble protein fraction resulted in the confident identification of 105 gut proteins, while the PBS-soluble fraction yielded an additional 37 R. microplus proteins. The results show an abundance of proteins involved in mitochondrial ATP synthesis, electron transport chain, protein synthesis, chaperone, antioxidant and protein folding and transport activities in midgut tissues of adult female ticks. Among the novel products identified were clathrin-adaptor protein, which is involved in the assembly of clathrin-coated vesicles, and membrane-associated trafficking proteins such as syntaxin 6 and surfeit 4. The observations allow the formulation of hypotheses regarding midgut physiology and will serve as a basis for future vaccine development and tick-host interaction research.
Vaccines
The negative impact of ticks and tick-borne diseases on animals and human health is driving research to discover novel targets affecting both vectors and pathogens. The salivary glands are involved in feeding and pathogen transmission, thus are considered as a compelling target to focus research. In this study, proteomics approach was used to characterize Rhipicephalus bursa sialoproteome in response to Babesia ovis infection and blood feeding. Two potential tick protective antigens were identified and its influence in tick biological parameters and pathogen infection was evaluated. Results demonstrate that the R. bursa sialoproteome is highly affected by feeding but infection is well tolerated by tick cells. The combination of both stimuli shifts the previous scenario and a more evident pathogen manipulation can be suggested. Knockdown of ub2n led to a significative increase of infection in tick salivary glands but a brusque decrease in the progeny, revealing its importance in the ...
Transmembrane proteins – Mining the cattle tick transcriptome
Ticks and Tick-borne Diseases, 2015
Managing the spread and load of pathogen-transmitting ticks is an important task worldwide. The cattle tick, Rhipicephalus microplus, not only impacts the economy through losses in dairy and meat production, but also raises concerns for human health in regards to the potential of certain transmitted pathogens becoming zoonotic. However, novel strategies to control R. microplus are hindered by lack of understanding tick biology and the discovery of suitable vaccine or acaricide targets. The importance of transmembrane proteins as vaccine targets are well known, as is the case in tick vaccines with Bm86 as antigen. In this study, we describe the localization and functional annotation of 878 putative transmembrane proteins. Thirty proteins could be confirmed in the R. microplus gut using LC-MS/MS analysis and their roles in tick biology are discussed. To the best of our knowledge, 19 targets have not been reported before in any proteomics study in various tick species and the possibility of using the identified proteins as targets for tick control are discussed. Although tissue expression of identified putative proteins through expansive proteomics is necessary, this study demonstrates the possibility of using bioinformatics for the identification of targets for further evaluation in tick control strategies. Protein identity Antigen name Species Source Protein type(s) Experimental host(s) Vaccine efficacy Reference 5'-nucleotidase 5'-nucleotidase (4F8) Rhipicephalus microplus Malpighian tubules Recombinant Cattle NSE Hope et al., 2010 Akirin rAKR Various Whole mosquito Recombinant Red deer 25-33% reduction in tick infestation Carreón et al. 2012 Angiotensin-converting enzyme Bm91 Rhipicephalus microplus Salivary glands Recombinant Cattle 6 and 8% reproductive efficiency and egg viability Lambertz et al., 2012 Angiotensin-converting enzyme Bm91 Rhipicephalus microplus Salivary glands Native Cattle ~37% reduction in eggs Riding et al., 1994 Aquaporin Aquaporin Rhipicephalus microplus Midgut Recombinant Cattle 75% and 68% Guerrero et al. 2014 Aspartic proteinase BYC Rhipicephalus microplus Eggs Native Cattle 14%and 36% da Silva Vaz et al., 1998; Seixas et al., 2012 Aspartic proteinase BYC Rhipicephalus microplus Eggs Recombinant Cattle 25% Leal et al., 2006; Seixas et al., 2012 Cement protein 64TRP Rhipicephalus appendiculatus Salivary glands Recombinant Guinea pigs ~62% mortality Trimnell et al., 2002 Cement protein 64TRP Rhipicephalus sangiuneus Salivary glands Recombinant Guinea pigs ~47% mortality Trimnell et al., 2005 Cement protein TrP64 Rhipicephalus appendiculatus N/i Recombinant Rabbits 43.9% Saimo et al. 2011 Component of vitellin GP80 Rhipicephalus microplus Various Native Sheep 68% Tellam et al., 2002 Component of vitellogenin, vitellin VIT87 Rhipicephalus microplus Eggs Native Sheep 68% Tellam et al., 2002 Elongation factor 1 alpha Ef1a Rhipicephalus microplus Various Recombinant Cattle 31% Almazán et al., 2012 Protein identity Antigen name Species Source Protein type(s) Experimental host(s) Vaccine efficacy Reference Extracellular matrix protein, Glycine-rich protein RH50 Rhipicephalus haemaphysaloides Salivary glands Recombinant Rabbits 30.5% mortality Zhou et al., 2006 Ferritin 2 RaFER2 Rhipicephalus annulatus Midgut Recombinant Cattle 72% Hajdusek et al., 2010 Ferritin 2 RmFER2 Rhipicephalus microplus Midgut Recombinant Cattle 64% Hajdusek et al., 2010 Glutathione S-transferase GST-Hl Rhipicephalus microplus Various Recombinant Cattle 57% Parizi et al., 2011 Glutathione S-transferase, Vitellin-degrading enzyme and Aspartic proteinase GST-Hl, VTDCE and BYC Rhipicephalus microplus Various Recombinant Cattle 35.3 to 61.6% protection against infestation Parizi et al., 2012 Mating factor, voraxinα Voraxinα Rhipicephalus appendiculatus Testis Recombinant Rabbits ~50% reduction in eggs Yamada et al., 2009 Metalloprotease BrRm-MP4 Rhipicephalus microplus Larvae Recombinant Cattle 60% Ali et al. 2015 Mucin BMA7 Rhipicephalus microplus Whole ticks, membrane fractions Native Cattle ~21% reduction in eggs McKenna et al., 1998 Ribosomal protein P0 pP0-KHL Rhipicephalus sanguineus Various Synthetic peptide Rabbits 90.25% Rodríguez-Mallon et al., 2012 Serpin-3 (RAS), Serpin-4 (RAS-4) and 36kDa immuno-dominant protein (RIM36) RAS-3, RAS-4 and RIM36 Rhipicephalus appendiculatus Salivary glands Recombinant Cattle ~27% mortality Imamura et al., 2008 Strain variant of Bm86 Bm95 Rhipicephalus microplus Midgut Recombinant Cattle 58-89% Garcia-Garcia et al., 2000 Strain variant of Bm86 Bm95 Rhipicephalus haemaphysaloides Midgut Recombinant Cattle 78.9-84.6% Sugumar et al., 2011 Protein identity Antigen name Species Source Protein type(s) Experimental host(s) Vaccine efficacy Reference Trypsin inhibitor BmLTI Rhipicephalus microplus Various Recombinant Cattle 32% Andreotti et al.,, 2012 Trypsin inhibitor BmTI Rhipicephalus microplus Salivary glands Native Cattle 72.8% Andreotti et al., 2002 Trypsin inhibitor BmTI-A Rhipicephalus microplus Salivary glands Synthetic Cattle ~18.4% Andreotti et al., 2007 Ubiquitin UBE Rhipicephalus annulatus Various Recombinant Cattle 15% and 22% depending construct preparation Almazán et al., 2010, Almazán et al., 2012 Ubiquitin UBE Rhipicephalus microplus Various Recombinant Cattle 55% Almazán et al., 2010 Unknown ARS antigen 1 Rhipicephalus microplus Midgut Recombinant Cattle 73-76%
Tick borne diseases in cattle Applications of proteomics to develop new generation vaccines
Tick-borne diseases (TBDs) affect 80% of the world's cattle population, hampering livestock production throughout the world. Livestock industry is important to rural populations not only as food supply, but also as a source of income. Tick control is usually achieved by using acaricides which are expensive, deleterious to the environment and can induce chemical resistance of vectors; the development of more effective and sustainable control methods is therefore required.
Identification of proteins expressed by Babesia bigemina kinetes
Parasites & Vectors
Background: Babesia bigemina is an apicomplexan parasite transovarially transmitted via Rhipicephalus ticks that infect red blood cells and causes bovine babesiosis, a poorly controlled severe acute disease in cattle. New methods of control are urgently needed, including the development of transmission blocking vaccines (TBV). Babesia bigemina reproduces sexually in the gut of adult female R. microplus upon acquisition following a blood meal. Sexual reproduction results in zygotes that infect gut epithelial cells to transform into kinete stage parasites, which invade tick ovaries and infects the egg mass. The subsequent tick generation transmits B. bigemina upon feeding on bovine hosts. An important limitation for developing novel TBV is that the pattern of protein expression in B. bigemina tick stages, such as the kinete stage, remain essentially uncharacterized. Results: We determined the protein expression profile of three B. bigemina putative tick stage candidates BbiKSP (BBBOND_0206730), CCp2 and CCp3. We found that BbiKSP expression was restricted to B. bigemina kinetes. CCp2 and CCp3, previously shown to be expressed by induced sexual stages, were also expressed by kinetes. Importantly, none of these proteins were expressed by B. bigemina blood stages. Conclusions: Babesia bigemina kinetes express BbiKSP, CCp2 and CCp3 proteins, therefore, these proteins may play important roles during B. bigemina development within tick hemolymph and may serve as potential candidate targets for the development of TBV.
Vaccines
Tick-borne diseases affecting humans and animals are on the rise worldwide. Vaccines constitute an effective control measure, but very few are available. We selected Lyme borreliosis, a bacterial infection transmitted by the hard tick Ixodes, to validate a new concept to identify vaccine candidates. This disease is the most common tick-borne disease in the Northern Hemisphere. Although attempts to develop a vaccine exist, none have been successfully marketed. In tick-borne diseases, the skin constitutes a very specific environment encountered by the pathogen during its co-inoculation with tick saliva. In a mouse model, we developed a proteomic approach to identify vaccine candidates in skin biopsies. We identified 30 bacterial proteins after syringe inoculation or tick inoculation of bacteria. Discovery proteomics using mass spectrometry might be used in various tick-borne diseases to identify pathogen proteins with early skin expression. It should help to better develop sub-unit va...