37 results on '"Ayllón N"'
Search Results
2. Tick-Pathogen Interactions and Vector Competence: Identification of Molecular Drivers for Tick-Borne Diseases
- Author
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de la Fuente, J., Antunes, S., Bonnet, S., Cabezas-Cruz, A., Domingos, A.G., Estrada-Peña, A., Johnson, N., Kocan, K.M., Mansfield, K.L., Nijhof, A.M., Papa, A., Rudenko, N., Villar, M., Alberdi, P., Torina, A., Ayllón, N., Vancova, M., Golovchenko, M., Grubhoffer, L., Caracappa, S., Fooks, A.R., Gortazar, C., and Rego, R.O.M.
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parasitic diseases ,bacterial infections and mycoses - Abstract
Ticks and the pathogens they transmit constitute a growing burden for human and animal health worldwide. Vector competence is a component of vectorial capacity and depends on genetic determinants affecting the ability of a vector to transmit a pathogen. These determinants affect traits such as tick-host-pathogen and susceptibility to pathogen infection. Therefore, the elucidation of the mechanisms involved in tick-pathogen interactions that affect vector competence is essential for the identification of molecular drivers for tick-borne diseases. In this review, we provide a comprehensive overview of tick-pathogen molecular interactions for bacteria, viruses, and protozoa affecting human and animal health. Additionally, the impact of tick microbiome on these interactions was considered. Results show that different pathogens evolved similar strategies such as manipulation of the immune response to infect vectors and facilitate multiplication and transmission. Furthermore, some of these strategies may be used by pathogens to infect both tick and mammalian hosts. Identification of interactions that promote tick survival, spread, and pathogen transmission provides the opportunity to disrupt these interactions and lead to a reduction in tick burden and the prevalence of tick-borne diseases. Targeting some of the similar mechanisms used by the pathogens for infection and transmission by ticks may assist in development of preventative strategies against multiple tick-borne diseases.
- Published
- 2017
3. Expression of heat shock proteins and subolesin affects stress responses, Anaplasma phagocytophilum infection and questing behaviour in the tick, Ixodes scapularis
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BUSBY, A. T., primary, AYLLÓN, N., additional, KOCAN, K. M., additional, BLOUIN, E. F., additional, DE LA FUENTE, G., additional, GALINDO, R. C., additional, VILLAR, M., additional, and DE LA FUENTE, J., additional
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- 2011
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4. Automatic pole-zero identification for multivariable large-signal stability analysis of RF and microwave circuits
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Anakabe, A., Ayllón, N., Juan María Collantes Metola, Mallet, A., Soubercaze-Pun, G., and Narendra, K.
5. Very High Frequency Power Switching: A Road Map To Envelope Tracking
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Delepaut Christophe and Ayllon Natanael
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Environmental sciences ,GE1-350 - Abstract
RF (Radio Frequency) GaN transistors may be used for power switching at frequencies of 30 MHz and above, thereby reaching voltage control bandwidth of critical interest for the so-called ET (Envelope Tracking) technique. ET is meant at optimising the efficiency of RF amplifiers by supplying them with a voltage adapted to their power level at any time, and is the subject of many R&D works worldwide. The present paper provides an overview on the different activities run at the European Space Agency so far in this context, and present the possible way-forward to embark ET on future spacecraft.
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- 2017
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6. Gene expression profile suggests that pigs (Sus scrofa) are susceptible to Anaplasma phagocytophilum but control infection
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Galindo Ruth C, Ayllón Nieves, Smrdel Katja, Boadella Mariana, Beltrán-Beck Beatriz, Mazariegos María, García Nerea, de la Lastra José M, Avsic-Zupanc Tatjana, Kocan Katherine M, Gortazar Christian, and de la Fuente José
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Anaplasmosis ,Genetics ,Pig ,Wild boar ,Genomics ,Immune response ,Infectious and parasitic diseases ,RC109-216 - Abstract
Abstract Background Anaplasma phagocytophilum infects a wide variety of hosts and causes granulocytic anaplasmosis in humans, horses and dogs and tick-borne fever in ruminants. Infection with A. phagocytophilum results in the modification of host gene expression and immune response. The objective of this research was to characterize gene expression in pigs (Sus scrofa) naturally and experimentally infected with A. phagocytophilum trying to identify mechanisms that help to explain low infection prevalence in this species. Results For gene expression analysis in naturally infected pigs, microarray hybridization was used. The expression of differentially expressed immune response genes was analyzed by real-time RT-PCR in naturally and experimentally infected pigs. Results suggested that A. phagocytophilum infection affected cytoskeleton rearrangement and increased both innate and adaptive immune responses by up regulation of interleukin 1 receptor accessory protein-like 1 (IL1RAPL1), T-cell receptor alpha chain (TCR-alpha), thrombospondin 4 (TSP-4) and Gap junction protein alpha 1 (GJA1) genes. Higher serum levels of IL-1 beta, IL-8 and TNF-alpha in infected pigs when compared to controls supported data obtained at the mRNA level. Conclusions These results suggested that pigs are susceptible to A. phagocytophilum but control infection, particularly through activation of innate immune responses, phagocytosis and autophagy. This fact may account for the low infection prevalence detected in pigs in some regions and thus their low or no impact as a reservoir host for this pathogen. These results advanced our understanding of the molecular mechanisms at the host-pathogen interface and suggested a role for newly reported genes in the protection of pigs against A. phagocytophilum.
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- 2012
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7. Functional genomics of the horn fly, Haematobia irritans (Linnaeus, 1758)
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Quiroz-Romero Héctor, Rosario-Cruz Rodrigo, Galindo Ruth C, Ayllón Nieves, Almazán Consuelo, Torres Lorena, and de la Fuente José
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Biotechnology ,TP248.13-248.65 ,Genetics ,QH426-470 - Abstract
Abstract Background The horn fly, Haematobia irritans (Linnaeus, 1758) (Diptera: Muscidae) is one of the most important ectoparasites of pastured cattle. Horn flies infestations reduce cattle weight gain and milk production. Additionally, horn flies are mechanical vectors of different pathogens that cause disease in cattle. The aim of this study was to conduct a functional genomics study in female horn flies using Expressed Sequence Tags (EST) analysis and RNA interference (RNAi). Results A cDNA library was made from whole abdominal tissues collected from partially fed adult female horn flies. High quality horn fly ESTs (2,160) were sequenced and assembled into 992 unigenes (178 contigs and 814 singlets) representing molecular functions such as serine proteases, cell metabolism, mitochondrial function, transcription and translation, transport, chromatin structure, vitellogenesis, cytoskeleton, DNA replication, cell response to stress and infection, cell proliferation and cell-cell interactions, intracellular trafficking and secretion, and development. Functional analyses were conducted using RNAi for the first time in horn flies. Gene knockdown by RNAi resulted in higher horn fly mortality (protease inhibitor functional group), reduced oviposition (vitellogenin, ferritin and vATPase groups) or both (immune response and 5'-NUC groups) when compared to controls. Silencing of ubiquitination ESTs did not affect horn fly mortality and ovisposition while gene knockdown in the ferritin and vATPse functional groups reduced mortality when compared to controls. Conclusions These results advanced the molecular characterization of this important ectoparasite and suggested candidate protective antigens for the development of vaccines for the control of horn fly infestations.
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- 2011
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8. Non-destructive 3D characterization of the blood vessel wall microstructure in different species and blood vessel types using contrast-enhanced microCT and comparison with synthetic vascular grafts.
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Leyssens L, Balcaen T, Pétré M, Béjar Ayllón N, El Aazmani W, de Pierpont A, Pyka G, Lacroix V, and Kerckhofs G
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- Humans, Rats, Animals, Swine, X-Ray Microtomography, Stents, Hafnium, Blood Vessel Prosthesis
- Abstract
To improve the current treatment for vascular diseases, such as vascular grafts, intravascular stents, and balloon angioplasty intervention, the evaluation of the native blood vessel microstructure in full 3D could be beneficial. For this purpose, we used contrast-enhanced X-ray microfocus computed tomography (CECT): a combination of X-ray microfocus computed tomography (microCT) and contrast-enhancing staining agents (CESAs) containing high atomic number elements. In this work, we performed a comparative study based on staining time and contrast-enhancement of 2 CESAs: Monolacunary and 1:2 Hafnium-substituted Wells-Dawson polyoxometalate (Mono-WD POM and Hf-WD POM, respectively) for imaging of the porcine aorta. After showing the advantages of Hf-WD POM in terms of contrast enhancement, we expanded our imaging to other species (rat, porcine, and human) and other types of blood vessels (porcine aorta, femoral artery, and vena cava), clearly indicating microstructural differences between different types of blood vessels and different species. We then showed the possibility to extract useful 3D quantitative information from the rat and porcine aortic wall, potentially to be used for computational modeling or for future design optimization of graft materials. Finally, a structural comparison with existing synthetic vascular grafts was made. This information will allow to better understand the in vivo functioning of native blood vessels and to improve the current disease treatments. STATEMENT OF SIGNIFICANCE: Synthetic vascular grafts, used as treatment for some cardiovascular diseases, still often fail clinically, potentially because of a mismatch in mechanical behaviour between the native blood vessel and the graft. To better understand the causes of this mismatch, we studied the full 3D microstructure of blood vessels. For this, we identified Hafnium-substituted Wells-Dawson polyoxometalate as contrast-enhancing staining agent to perform contrast-enhanced X-ray microfocus computed tomography. This technique allowed to show important differences in the microstructure of different types of blood vessels and in different species, as well as with that of synthetic grafts. This information can lead to a better understanding of the functioning of blood vessels and will allow to improve current disease treatments, such as vascular grafts., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)
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- 2023
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9. Anaplasma phagocytophilum MSP4 and HSP70 Proteins Are Involved in Interactions with Host Cells during Pathogen Infection.
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Contreras M, Alberdi P, Mateos-Hernández L, Fernández de Mera IG, García-Pérez AL, Vancová M, Villar M, Ayllón N, Cabezas-Cruz A, Valdés JJ, Stuen S, Gortazar C, and de la Fuente J
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- Anaplasma phagocytophilum genetics, Animals, Bacterial Proteins genetics, Ehrlichiosis microbiology, HSP70 Heat-Shock Proteins genetics, Host-Pathogen Interactions, Membrane Proteins genetics, Sheep, Anaplasma phagocytophilum metabolism, Bacterial Proteins metabolism, Ehrlichiosis veterinary, HSP70 Heat-Shock Proteins metabolism, Membrane Proteins metabolism, Sheep Diseases microbiology
- Abstract
Anaplasma phagocytophilum transmembrane and surface proteins play a role during infection and multiplication in host neutrophils and tick vector cells. Recently, A. phagocytophilum Major surface protein 4 (MSP4) and Heat shock protein 70 (HSP70) were shown to be localized on the bacterial membrane, with a possible role during pathogen infection in ticks. In this study, we hypothesized that A. phagocytophilum MSP4 and HSP70 have similar functions in tick-pathogen and host-pathogen interactions. To address this hypothesis, herein we characterized the role of these bacterial proteins in interaction and infection of vertebrate host cells. The results showed that A. phagocytophilum MSP4 and HSP70 are involved in host-pathogen interactions, with a role for HSP70 during pathogen infection. The analysis of the potential protective capacity of MSP4 and MSP4-HSP70 antigens in immunized sheep showed that MSP4-HSP70 was only partially protective against pathogen infection. This limited protection may be associated with several factors, including the recognition of non-protective epitopes by IgG in immunized lambs. Nevertheless, these antigens may be combined with other candidate protective antigens for the development of vaccines for the control of human and animal granulocytic anaplasmosis. Focusing on the characterization of host protective immune mechanisms and protein-protein interactions at the host-pathogen interface may lead to the discovery and design of new effective protective antigens.
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- 2017
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10. Comparative Proteomics Reveals Differences in Host-Pathogen Interaction between Infectious and Commensal Relationship with Campylobacter jejuni .
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Ayllón N, Jiménez-Marín Á, Argüello H, Zaldívar-López S, Villar M, Aguilar C, Moreno A, De La Fuente J, and Garrido JJ
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- Animals, Bacterial Proteins metabolism, Campylobacter jejuni isolation & purification, Campylobacter jejuni metabolism, Cell Cycle, Cell Line, Cell Movement, Chickens microbiology, Clathrin pharmacology, Endocytosis, Epithelial Cells immunology, Epithelial Cells microbiology, Gene Expression Regulation, Bacterial, Humans, Intestines microbiology, Proteome analysis, Reactive Oxygen Species, Signal Transduction, Swine, Virulence Factors metabolism, Campylobacter Infections microbiology, Campylobacter jejuni pathogenicity, Host-Pathogen Interactions physiology, Proteomics methods, Symbiosis physiology
- Abstract
Campylobacter jejuni is the leading food-borne poisoning in industrialized countries. While the bacteria causes disease in humans, it merely colonizes the gut in poultry or pigs, where seems to establish a commensal relationship. Until now, few studies have been conducted to elucidate the relationship between C. jejuni and its different hosts. In this work, a comparative proteomics approach was used to identify the underlying mechanisms involved in the divergent outcome following C. jejuni infection in human and porcine host. Human (INT-407) and porcine (IPEC-1) intestinal cell lines were infected by C. jejuni for 3 h (T3h) and 24 h (T24h). C. jejuni infection prompted an intense inflammatory response at T3h in human intestinal cells, mainly characterized by expression of proteins involved in cell spreading, cell migration and promotion of reactive oxygen species (ROS). Proteomic analysis evidenced significantly regulated biofunctions in human cells related with engulfment and endocytosis, and supported by canonical pathways associated to infection such as caveolar- and clathrin-mediated endocytosis signaling. In porcine IPEC-1 cells, inflammatory response as well as signaling pathways that control cellular functions such as cell migration, endocytosis and cell cycle progression resulted downregulated. These differences in the host response to infection were supported by the different pattern of adhesion and invasion proteins expressed by C. jejuni in human and porcine cells. No marked differences in expression of virulence factors involved in adaptive response and iron acquisition functions were observed. Therefore, the results of this study suggest that both host and pathogen factors are responsible for commensal or infectious character of C. jejuni in different hosts.
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- 2017
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11. Tick-Pathogen Interactions and Vector Competence: Identification of Molecular Drivers for Tick-Borne Diseases.
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de la Fuente J, Antunes S, Bonnet S, Cabezas-Cruz A, Domingos AG, Estrada-Peña A, Johnson N, Kocan KM, Mansfield KL, Nijhof AM, Papa A, Rudenko N, Villar M, Alberdi P, Torina A, Ayllón N, Vancova M, Golovchenko M, Grubhoffer L, Caracappa S, Fooks AR, Gortazar C, and Rego ROM
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- Animals, Arachnid Vectors parasitology, Humans, Ticks microbiology, Ticks parasitology, Ticks virology, Arachnid Vectors microbiology, Arachnid Vectors virology, Disease Transmission, Infectious, Host-Pathogen Interactions, Tick-Borne Diseases epidemiology, Ticks physiology
- Abstract
Ticks and the pathogens they transmit constitute a growing burden for human and animal health worldwide. Vector competence is a component of vectorial capacity and depends on genetic determinants affecting the ability of a vector to transmit a pathogen. These determinants affect traits such as tick-host-pathogen and susceptibility to pathogen infection. Therefore, the elucidation of the mechanisms involved in tick-pathogen interactions that affect vector competence is essential for the identification of molecular drivers for tick-borne diseases. In this review, we provide a comprehensive overview of tick-pathogen molecular interactions for bacteria, viruses, and protozoa affecting human and animal health. Additionally, the impact of tick microbiome on these interactions was considered. Results show that different pathogens evolved similar strategies such as manipulation of the immune response to infect vectors and facilitate multiplication and transmission. Furthermore, some of these strategies may be used by pathogens to infect both tick and mammalian hosts. Identification of interactions that promote tick survival, spread, and pathogen transmission provides the opportunity to disrupt these interactions and lead to a reduction in tick burden and the prevalence of tick-borne diseases. Targeting some of the similar mechanisms used by the pathogens for infection and transmission by ticks may assist in development of preventative strategies against multiple tick-borne diseases.
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- 2017
- Full Text
- View/download PDF
12. The intracellular bacterium Anaplasma phagocytophilum selectively manipulates the levels of vertebrate host proteins in the tick vector Ixodes scapularis.
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Villar M, López V, Ayllón N, Cabezas-Cruz A, López JA, Vázquez J, Alberdi P, and de la Fuente J
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- Animals, Arthropod Proteins genetics, Arthropod Proteins metabolism, Gene Expression Regulation physiology, Heat-Shock Proteins genetics, Heat-Shock Proteins metabolism, Host-Pathogen Interactions, RNA, Messenger genetics, RNA, Messenger metabolism, Anaplasma phagocytophilum physiology, Ixodes metabolism, Ixodes microbiology, Sheep metabolism
- Abstract
Background: The intracellular bacteria Anaplasma phagocytophilum are emerging zoonotic pathogens affecting human and animal health, and a good model for the study of tick-host-pathogen interactions. This tick-borne pathogen is transmitted by Ixodes scapularis in the United States where it causes human granulocytic anaplasmosis. Tick midguts and salivary glands play a major role during tick feeding and development, and in pathogen acquisition, multiplication and transmission. Vertebrate host proteins are found in tick midguts after feeding and have been described in the salivary glands of fed and unfed ticks, suggesting a role for these proteins during tick feeding and development. Furthermore, recent results suggested the hypothesis that pathogen infection affects tick metabolic processes to modify host protein digestion and persistence in the tick with possible implications for tick physiology and pathogen life-cycle., Methods: To address this hypothesis, herein we used I. scapularis female ticks fed on uninfected and A. phagocytophilum-infected sheep to characterize host protein content in midguts and salivary glands by proteomic analysis of tick tissues., Results: The results evidenced a clear difference in the host protein content between tick midguts and salivary glands in response to infection suggesting that A. phagocytophilum selectively manipulates the levels of vertebrate host proteins in ticks in a tissue-specific manner to facilitate pathogen infection, multiplication and transmission while preserving tick feeding and development. The mechanisms by which A. phagocytophilum manipulates the levels of vertebrate host proteins are not known, but the results obtained here suggested that it might include the modification of proteolytic pathways., Conclusions: The results of this study provided evidence to support that A. phagocytophilum affect tick proteolytic pathways to selectively manipulate the levels of vertebrate host proteins in a tissue-specific manner to increase tick vector capacity. Investigating the biological relevance of host proteins in tick biology and pathogen infection and the mechanisms used by A. phagocytophilum to manipulate host protein content is essential to advance our knowledge of tick-host-pathogen molecular interactions. These results have implications for the identification of new targets for the development of vaccines for the control of tick-borne diseases.
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- 2016
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13. Tick-Host-Pathogen Interactions: Conflict and Cooperation.
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de la Fuente J, Villar M, Cabezas-Cruz A, Estrada-Peña A, Ayllón N, and Alberdi P
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- Animals, Arthropod Proteins immunology, Humans, Inflammation immunology, Host-Pathogen Interactions immunology, Tick Infestations immunology, Ticks immunology
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- 2016
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14. Anaplasma phagocytophilum increases the levels of histone modifying enzymes to inhibit cell apoptosis and facilitate pathogen infection in the tick vector Ixodes scapularis.
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Cabezas-Cruz A, Alberdi P, Ayllón N, Valdés JJ, Pierce R, Villar M, and de la Fuente J
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- Animals, Apoptosis genetics, Cell Line, Histones metabolism, Host-Pathogen Interactions genetics, Humans, Insect Vectors genetics, Ixodes genetics, RNA, Messenger biosynthesis, RNA, Messenger genetics, Transcriptome genetics, p300-CBP Transcription Factors biosynthesis, p300-CBP Transcription Factors genetics, Anaplasma phagocytophilum genetics, Epigenesis, Genetic, Histone Code genetics, Histones genetics
- Abstract
Epigenetic mechanisms have not been characterized in ticks despite their importance as vectors of human and animal diseases worldwide. The objective of this study was to characterize the histones and histone modifying enzymes (HMEs) of the tick vector Ixodes scapularis and their role during Anaplasma phagocytophilum infection. We first identified 5 histones and 34 HMEs in I. scapularis in comparison with similar proteins in model organisms. Then, we used transcriptomic and proteomic data to analyze the mRNA and protein levels of I. scapularis histones and HMEs in response to A. phagocytophilum infection of tick tissues and cultured cells. Finally, selected HMEs were functionally characterized by pharmacological studies in cultured tick cells. The results suggest that A. phagocytophilum manipulates tick cell epigenetics to increase I. scapularis p300/CBP, histone deacetylase, and Sirtuin levels, resulting in an inhibition of cell apoptosis that in turn facilitates pathogen infection and multiplication. These results also suggest that a compensatory mechanism might exist by which A. phagocytophilum manipulates tick HMEs to regulate transcription and apoptosis in a tissue-specific manner to facilitate infection, but preserving tick fitness to guarantee survival of both pathogens and ticks. Our study also indicates that the pathogen manipulates arthropod and vertebrate cell epigenetics in similar ways to inhibit the host response to infection. Epigenetic regulation of tick biological processes is an essential element of the infection by A. phagocytophilum and the study of the mechanisms and principal actors involved is likely to provide clues for the development of anti-tick drugs and vaccines.
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- 2016
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15. Tissue-Specific Signatures in the Transcriptional Response to Anaplasma phagocytophilum Infection of Ixodes scapularis and Ixodes ricinus Tick Cell Lines.
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Alberdi P, Mansfield KL, Manzano-Román R, Cook C, Ayllón N, Villar M, Johnson N, Fooks AR, and de la Fuente J
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- Animals, Base Sequence, Cell Line, Gene Expression Regulation genetics, Sequence Analysis, RNA, Transcription, Genetic genetics, Anaplasma phagocytophilum pathogenicity, Apoptosis genetics, Host-Pathogen Interactions genetics, Ixodes microbiology, RNA genetics
- Abstract
Anaplasma phagocytophilum are transmitted by Ixodes spp. ticks and have become one of the most common and relevant tick-borne pathogens due to their impact on human and animal health. Recent results have increased our understanding of the molecular interactions between Ixodes scapularis and A. phagocytophilum through the demonstration of tissue-specific molecular pathways that ensure pathogen infection, development and transmission by ticks. However, little is known about the Ixodes ricinus genes and proteins involved in the response to A. phagocytophilum infection. The tick species I. scapularis and I. ricinus are evolutionarily closely related and therefore similar responses are expected in A. phagocytophilum-infected cells. However, differences may exist between I. scapularis ISE6 and I. ricinus IRE/CTVM20 tick cells associated with tissue-specific signatures of these cell lines. To address this hypothesis, the transcriptional response to A. phagocytophilum infection was characterized by RNA sequencing and compared between I. scapularis ISE6 and I. ricinus IRE/CTVM20 tick cell lines. The transcriptional response to infection of I. scapularis ISE6 cells resembled that of tick hemocytes while the response in I. ricinus IRE/CTVM20 cells was more closely related to that reported previously in infected tick midguts. The inhibition of cell apoptosis by A. phagocytophilum appears to be a key adaptation mechanism to facilitate infection of both vertebrate and tick cells and was used to investigate further the tissue-specific response of tick cell lines to pathogen infection. The results supported a role for the intrinsic pathway in the inhibition of cell apoptosis by A. phagocytophilum infection of I. scapularis ISE6 cells. In contrast, the results in I. ricinus IRE/CTVM20 cells were similar to those obtained in tick midguts and suggested a role for the JAK/STAT pathway in the inhibition of apoptosis in tick cells infected with A. phagocytophilum. Nevertheless, tick cell lines were derived from embryonated eggs and may contain various cell populations with different morphology and behavior that could affect transcriptional response to infection. These results suggested tissue-specific signatures in I. scapularis ISE6 and I. ricinus IRE/CTVM20 tick cell line response to A. phagocytophilum infection that support their use as models for the study of tick-pathogen interactions.
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- 2016
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16. Genomic insights into the Ixodes scapularis tick vector of Lyme disease.
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Gulia-Nuss M, Nuss AB, Meyer JM, Sonenshine DE, Roe RM, Waterhouse RM, Sattelle DB, de la Fuente J, Ribeiro JM, Megy K, Thimmapuram J, Miller JR, Walenz BP, Koren S, Hostetler JB, Thiagarajan M, Joardar VS, Hannick LI, Bidwell S, Hammond MP, Young S, Zeng Q, Abrudan JL, Almeida FC, Ayllón N, Bhide K, Bissinger BW, Bonzon-Kulichenko E, Buckingham SD, Caffrey DR, Caimano MJ, Croset V, Driscoll T, Gilbert D, Gillespie JJ, Giraldo-Calderón GI, Grabowski JM, Jiang D, Khalil SMS, Kim D, Kocan KM, Koči J, Kuhn RJ, Kurtti TJ, Lees K, Lang EG, Kennedy RC, Kwon H, Perera R, Qi Y, Radolf JD, Sakamoto JM, Sánchez-Gracia A, Severo MS, Silverman N, Šimo L, Tojo M, Tornador C, Van Zee JP, Vázquez J, Vieira FG, Villar M, Wespiser AR, Yang Y, Zhu J, Arensburger P, Pietrantonio PV, Barker SC, Shao R, Zdobnov EM, Hauser F, Grimmelikhuijzen CJP, Park Y, Rozas J, Benton R, Pedra JHF, Nelson DR, Unger MF, Tubio JMC, Tu Z, Robertson HM, Shumway M, Sutton G, Wortman JR, Lawson D, Wikel SK, Nene VM, Fraser CM, Collins FH, Birren B, Nelson KE, Caler E, and Hill CA
- Subjects
- Animals, Gene Expression Profiling, Genomics, Lyme Disease transmission, Oocytes, Xenopus laevis, Anaplasma phagocytophilum, Arachnid Vectors genetics, Genome genetics, Ixodes genetics, Ligand-Gated Ion Channels genetics
- Abstract
Ticks transmit more pathogens to humans and animals than any other arthropod. We describe the 2.1 Gbp nuclear genome of the tick, Ixodes scapularis (Say), which vectors pathogens that cause Lyme disease, human granulocytic anaplasmosis, babesiosis and other diseases. The large genome reflects accumulation of repetitive DNA, new lineages of retro-transposons, and gene architecture patterns resembling ancient metazoans rather than pancrustaceans. Annotation of scaffolds representing ∼57% of the genome, reveals 20,486 protein-coding genes and expansions of gene families associated with tick-host interactions. We report insights from genome analyses into parasitic processes unique to ticks, including host 'questing', prolonged feeding, cuticle synthesis, blood meal concentration, novel methods of haemoglobin digestion, haem detoxification, vitellogenesis and prolonged off-host survival. We identify proteins associated with the agent of human granulocytic anaplasmosis, an emerging disease, and the encephalitis-causing Langat virus, and a population structure correlated to life-history traits and transmission of the Lyme disease agent.
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- 2016
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17. Integrated Metabolomics, Transcriptomics and Proteomics Identifies Metabolic Pathways Affected by Anaplasma phagocytophilum Infection in Tick Cells.
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Villar M, Ayllón N, Alberdi P, Moreno A, Moreno M, Tobes R, Mateos-Hernández L, Weisheit S, Bell-Sakyi L, and de la Fuente J
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- Animals, Cell Line, Ehrlichiosis genetics, Ehrlichiosis metabolism, Endoplasmic Reticulum genetics, Endoplasmic Reticulum metabolism, Glucose metabolism, Host-Pathogen Interactions, Metabolic Networks and Pathways, Systems Biology methods, Anaplasma phagocytophilum physiology, Ehrlichiosis veterinary, Metabolomics methods, Proteomics methods, Ticks microbiology
- Abstract
Anaplasma phagocytophilum is an emerging zoonotic pathogen that causes human granulocytic anaplasmosis. These intracellular bacteria establish infection by affecting cell function in both the vertebrate host and the tick vector, Ixodes scapularis. Previous studies have characterized the tick transcriptome and proteome in response to A. phagocytophilum infection. However, in the postgenomic era, the integration of omics datasets through a systems biology approach allows network-based analyses to describe the complexity and functionality of biological systems such as host-pathogen interactions and the discovery of new targets for prevention and control of infectious diseases. This study reports the first systems biology integration of metabolomics, transcriptomics, and proteomics data to characterize essential metabolic pathways involved in the tick response to A. phagocytophilum infection. The ISE6 tick cells used in this study constitute a model for hemocytes involved in pathogen infection and immune response. The results showed that infection affected protein processing in endoplasmic reticulum and glucose metabolic pathways in tick cells. These results supported tick-Anaplasma co-evolution by providing new evidence of how tick cells limit pathogen infection, while the pathogen benefits from the tick cell response to establish infection. Additionally, ticks benefit from A. phagocytophilum infection by increasing survival while pathogens guarantee transmission. The results suggested that A. phagocytophilum induces protein misfolding to limit the tick cell response and facilitate infection but requires protein degradation to prevent ER stress and cell apoptosis to survive in infected cells. Additionally, A. phagocytophilum may benefit from the tick cell's ability to limit bacterial infection through PEPCK inhibition leading to decreased glucose metabolism, which also results in the inhibition of cell apoptosis that increases infection of tick cells. These results support the use of this experimental approach to systematically identify cell pathways and molecular mechanisms involved in tick-pathogen interactions. Data are available via ProteomeXchange with identifier PXD002181., (© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.)
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- 2015
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18. Identification and Characterization of Anaplasma phagocytophilum Proteins Involved in Infection of the Tick Vector, Ixodes scapularis.
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Villar M, Ayllón N, Kocan KM, Bonzón-Kulichenko E, Alberdi P, Blouin EF, Weisheit S, Mateos-Hernández L, Cabezas-Cruz A, Bell-Sakyi L, Vancová M, Bílý T, Meyer DF, Sterba J, Contreras M, Rudenko N, Grubhoffer L, Vázquez J, and de la Fuente J
- Subjects
- Anaplasma phagocytophilum, Animals, Bacterial Proteins metabolism, Chaperonin 60 metabolism, Female, Gastrointestinal Tract microbiology, Gene Expression Profiling, Gene Expression Regulation, HSP70 Heat-Shock Proteins metabolism, Host-Pathogen Interactions, Membrane Proteins metabolism, Molecular Sequence Annotation, Proteome metabolism, Salivary Glands microbiology, Signal Transduction, Stress, Physiological, Bacterial Proteins genetics, Chaperonin 60 genetics, HSP70 Heat-Shock Proteins genetics, Ixodes microbiology, Membrane Proteins genetics, Proteome genetics
- Abstract
Anaplasma phagocytophilum is an emerging zoonotic pathogen transmitted by Ixodes scapularis that causes human granulocytic anaplasmosis. Here, a high throughput quantitative proteomics approach was used to characterize A. phagocytophilum proteome during rickettsial multiplication and identify proteins involved in infection of the tick vector, I. scapularis. The first step in this research was focused on tick cells infected with A. phagocytophilum and sampled at two time points containing 10-15% and 65-71% infected cells, respectively to identify key bacterial proteins over-represented in high percentage infected cells. The second step was focused on adult female tick guts and salivary glands infected with A. phagocytophilum to compare in vitro results with those occurring during bacterial infection in vivo. The results showed differences in the proteome of A. phagocytophilum in infected ticks with higher impact on protein synthesis and processing than on bacterial replication in tick salivary glands. These results correlated well with the developmental cycle of A. phagocytophilum, in which cells convert from an intracellular reticulated, replicative form to the nondividing infectious dense-core form. The analysis of A. phagocytophilum differentially represented proteins identified stress response (GroEL, HSP70) and surface (MSP4) proteins that were over-represented in high percentage infected tick cells and salivary glands when compared to low percentage infected cells and guts, respectively. The results demonstrated that MSP4, GroEL and HSP70 interact and bind to tick cells, thus playing a role in rickettsia-tick interactions. The most important finding of these studies is the increase in the level of certain bacterial stress response and surface proteins in A. phagocytophilum-infected tick cells and salivary glands with functional implication in tick-pathogen interactions. These results gave a new dimension to the role of these stress response and surface proteins during A. phagocytophilum infection in ticks. Characterization of Anaplasma proteome contributes information on host-pathogen interactions and provides targets for development of novel control strategies for pathogen infection and transmission.
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- 2015
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19. Bacterial membranes enhance the immunogenicity and protective capacity of the surface exposed tick Subolesin-Anaplasma marginale MSP1a chimeric antigen.
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Contreras M, Moreno-Cid JA, Domingos A, Canales M, Díez-Delgado I, Pérez de la Lastra JM, Sánchez E, Merino O, Zavala RL, Ayllón N, Boadella M, Villar M, Gortázar C, and de la Fuente J
- Subjects
- Adjuvants, Immunologic, Animals, Antibodies blood, Cell Membrane chemistry, Escherichia coli, Female, Mice, Mice, Inbred BALB C, Rabbits, Swine, Anaplasma marginale immunology, Antigens immunology, Arthropod Proteins immunology, Bacterial Outer Membrane Proteins immunology, Rhipicephalus immunology, Vaccines immunology
- Abstract
Ticks are vectors of diseases that affect humans and animals worldwide. Tick vaccines have been proposed as a cost-effective and environmentally sound alternative for tick control. Recently, the Rhipicephalus microplus Subolesin (SUB)-Anaplasma marginale MSP1a chimeric antigen was produced in Escherichia coli as membrane-bound and exposed protein and used to protect vaccinated cattle against tick infestations. In this research, lipidomics and proteomics characterization of the E. coli membrane-bound SUB-MSP1a antigen showed the presence of components with potential adjuvant effect. Furthermore, vaccination with membrane-free SUB-MSP1a and bacterial membranes containing SUB-MSP1a showed that bacterial membranes enhance the immunogenicity of the SUB-MSP1a antigen in animal models. R. microplus female ticks were capillary-fed with sera from pigs orally immunized with membrane-free SUB, membrane bound SUB-MSP1a and saline control. Ticks ingested antibodies added to the blood meal and the effect of these antibodies on reduction of tick weight was shown for membrane bound SUB-MSP1a but not SUB when compared to control. Using the simple and cost-effective process developed for the purification of membrane-bound SUB-MSP1a, endotoxin levels were within limits accepted for recombinant vaccines. These results provide further support for the development of tick vaccines using E. coli membranes exposing chimeric antigens such as SUB-MSP1a., (Copyright © 2015 Elsevier GmbH. All rights reserved.)
- Published
- 2015
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20. Infection of Ixodes spp. tick cells with different Anaplasma phagocytophilum isolates induces the inhibition of apoptotic cell death.
- Author
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Alberdi P, Ayllón N, Cabezas-Cruz A, Bell-Sakyi L, Zweygarth E, Stuen S, and de la Fuente J
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- Animals, Caspase 9 genetics, Caspase 9 metabolism, Cell Line, DNA, Bacterial genetics, Gene Expression Regulation, Enzymologic, Hexokinase genetics, Hexokinase metabolism, Ixodes embryology, Phylogeny, Anaplasma phagocytophilum physiology, Apoptosis physiology, Ixodes cytology
- Abstract
Anaplasma phagocytophilum is an intracellular rickettsial pathogen transmitted by Ixodes spp. ticks, which causes granulocytic anaplasmosis in humans, horses and dogs and tick-borne fever (TBF) in ruminants. In the United States, human granulocytic anaplasmosis (HGA) is highly prevalent while TBF has not been reported. However, in Europe the situation is the opposite, with high prevalence for TBF in sheep and low prevalence of HGA. The origin of these differences has not been identified and our hypothesis is that different A. phagocytophilum isolates impact differently on tick vector capacity through inhibition of apoptosis to establish infection of the tick vector. In this study we used three different isolates of A. phagocytophilum of human, canine and ovine origin to infect the Ixodes ricinus-derived cell line IRE/CTVM20 and the Ixodes scapularis-derived cell line ISE6 in order to characterize the effect of infection on the level of tick cell apoptosis. Inhibition of apoptosis was observed by flow cytometry as early as 24h post-infection for both tick cell lines and all three isolates of A. phagocytophilum, suggesting that pathogen infection inhibits apoptotic pathways to facilitate infection independently of the origin of the A. phagocytophilum isolate and tick vector species. However, infection with A. phagocytophilum isolates inhibited the intrinsic apoptosis pathway at different levels in I. scapularis and I. ricinus cells. These results suggested an impact of vector-pathogen co-evolution on the adaptation of A. phagocytophilum isolates to grow in tick cells as each isolate grew better in the tick cell line derived from its natural vector species. These results increase our understanding of the mechanisms of A. phagocytophilum infection and multiplication and suggest that multiple mechanisms may affect disease prevalence in different geographical regions., (Copyright © 2015 Elsevier GmbH. All rights reserved.)
- Published
- 2015
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21. Nuclease Tudor-SN Is Involved in Tick dsRNA-Mediated RNA Interference and Feeding but Not in Defense against Flaviviral or Anaplasma phagocytophilum Rickettsial Infection.
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Ayllón N, Naranjo V, Hajdušek O, Villar M, Galindo RC, Kocan KM, Alberdi P, Šíma R, Cabezas-Cruz A, Rückert C, Bell-Sakyi L, Kazimírová M, Havlíková S, Klempa B, Kopáček P, and de la Fuente J
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- Amino Acid Sequence, Animals, Cell Line, Conserved Sequence, Cricetinae, Ixodes parasitology, Ixodes virology, Molecular Sequence Data, Nuclear Proteins metabolism, Phylogeny, Transcriptome, Anaplasma phagocytophilum pathogenicity, Flavivirus pathogenicity, Ixodes genetics, Nuclear Proteins genetics, RNA Interference
- Abstract
Tudor staphylococcal nuclease (Tudor-SN) and Argonaute (Ago) are conserved components of the basic RNA interference (RNAi) machinery with a variety of functions including immune response and gene regulation. The RNAi machinery has been characterized in tick vectors of human and animal diseases but information is not available on the role of Tudor-SN in tick RNAi and other cellular processes. Our hypothesis is that tick Tudor-SN is part of the RNAi machinery and may be involved in innate immune response and other cellular processes. To address this hypothesis, Ixodes scapularis and I. ricinus ticks and/or cell lines were used to annotate and characterize the role of Tudor-SN in dsRNA-mediated RNAi, immune response to infection with the rickettsia Anaplasma phagocytophilum and the flaviviruses TBEV or LGTV and tick feeding. The results showed that Tudor-SN is conserved in ticks and involved in dsRNA-mediated RNAi and tick feeding but not in defense against infection with the examined viral and rickettsial pathogens. The effect of Tudor-SN gene knockdown on tick feeding could be due to down-regulation of genes that are required for protein processing and blood digestion through a mechanism that may involve selective degradation of dsRNAs enriched in G:U pairs that form as a result of adenosine-to-inosine RNA editing. These results demonstrated that Tudor-SN plays a role in tick RNAi pathway and feeding but no strong evidence for a role in innate immune responses to pathogen infection was found.
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- 2015
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22. Identification and characterization of a novel tick-borne flavivirus subtype in goats (Capra hircus) in Spain.
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Mansfield KL, Morales AB, Johnson N, Ayllón N, Höfle U, Alberdi P, Fernández de Mera IG, Marín JF, Gortázar C, de la Fuente J, and Fooks AR
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- Animals, Cluster Analysis, Encephalitis Viruses, Tick-Borne genetics, Encephalitis, Tick-Borne virology, Ireland, Molecular Sequence Data, Phylogeny, Sequence Homology, Spain, Encephalitis Viruses, Tick-Borne isolation & purification, Encephalitis, Tick-Borne veterinary, Genome, Viral, Goat Diseases virology, Goats virology, RNA, Viral genetics, Sequence Analysis, DNA
- Abstract
In 2011, a neurological disease was reported in a herd of goats (Capra hircus) in Asturias, Spain. Initial sequencing identified the causative agent as louping ill virus (LIV). Subsequently, with the application of whole genome sequencing and phylogenetic analysis, empirical data demonstrates that the LIV-like virus detected is significantly divergent from LIV and Spanish sheep encephalitis virus (SSEV). This virus encoded an amino acid sequence motif at the site of a previously identified marker for differentiating tick-borne flaviviruses that was shared with a virus previously isolated in Ireland in 1968. The significance of these observations reflects the diversity of tick-borne flaviviruses in Europe. These data also contribute to our knowledge of the evolution of tick-borne flaviviruses and could reflect the movement of viruses throughout Europe. Based on these observations, the proposed name for this virus is Spanish goat encephalitis virus (SGEV), to distinguish it from SSEV.
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- 2015
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23. Molecular and immunological characterization of three strains of Anaplasma marginale grown in cultured tick cells.
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Lis K, Fernández de Mera IG, Popara M, Cabezas-Cruz A, Ayllón N, Zweygarth E, Passos LM, Broniszewska M, Villar M, Kocan KM, Ribeiro MF, Pfister K, and de la Fuente J
- Subjects
- Anaplasma marginale classification, Anaplasma marginale genetics, Anaplasma marginale growth & development, Anaplasmosis microbiology, Animals, Antigenic Variation, Brazil, Cattle, Cattle Diseases microbiology, Conserved Sequence, Molecular Sequence Data, Phylogeny, United States, Anaplasma marginale isolation & purification, Anaplasmosis immunology, Arachnid Vectors microbiology, Bacterial Proteins genetics, Bacterial Proteins immunology, Cattle Diseases immunology, Ticks microbiology
- Abstract
Anaplasma marginale is an economically important tick-borne pathogen of cattle that causes bovine anaplasmosis. A wide range of geographic strains of A. marginale have been isolated from cattle, several of which have been characterized using genomics and proteomics. While many of these strains have been propagated in tick lines, comparative analyses after propagation in tick cells have not been reported. The overall purpose of this research therefore was to compare the degree of conservation of selected genes after propagation in tick cell culture among A. marginale strains from the U.S. (the Virginia strain) and Brazil (UFMG1 and UFMG2 strains). The genes studied herein included those which encode the proteins HSP70 and SODB involved in heat shock and stress responses, respectively, and two genes that encode major surface proteins MSP4 and MSP5. Strain identities were first confirmed by sequencing the tandem repeats of the msp1a gene which encodes for the adhesin, MSP1a. The results of these studies demonstrated that the genes encoding for both stress response and heat shock proteins were highly conserved among the three A. marginale strains. Antibodies specific for MSP4, MSP5, SODB and HSP70 proteins were used to further characterize the A. marginale strains, and they reacted with all of these strains propagated in tick cell culture, providing further evidence for antigenic conservation. Although antigenic differences were not found among the three A. marginale strains, multi-locus sequence analysis (MLSA) performed with nucleotide sequences of these genes demonstrated that the A. marginale Brazilian and U.S. strains fall in different clades. These results showed that phylogenetically distant strains of A. marginale are antigenically conserved, even after several in vitro passages, supporting the use of some of the above conserved proteins as candidates for universal vaccines., (Copyright © 2015 Elsevier GmbH. All rights reserved.)
- Published
- 2015
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24. Systems biology of tissue-specific response to Anaplasma phagocytophilum reveals differentiated apoptosis in the tick vector Ixodes scapularis.
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Ayllón N, Villar M, Galindo RC, Kocan KM, Šíma R, López JA, Vázquez J, Alberdi P, Cabezas-Cruz A, Kopáček P, and de la Fuente J
- Subjects
- Anaplasma phagocytophilum pathogenicity, Anaplasmosis microbiology, Anaplasmosis transmission, Animals, Cell Differentiation genetics, Female, Gene Expression Regulation, Humans, Insect Vectors genetics, Insect Vectors microbiology, Ixodes microbiology, Organ Specificity, RNA Interference, Salivary Glands metabolism, Salivary Glands microbiology, Signal Transduction genetics, Transcriptome genetics, Anaplasma phagocytophilum genetics, Anaplasmosis genetics, Apoptosis genetics, Systems Biology
- Abstract
Anaplasma phagocytophilum is an emerging pathogen that causes human granulocytic anaplasmosis. Infection with this zoonotic pathogen affects cell function in both vertebrate host and the tick vector, Ixodes scapularis. Global tissue-specific response and apoptosis signaling pathways were characterized in I. scapularis nymphs and adult female midguts and salivary glands infected with A. phagocytophilum using a systems biology approach combining transcriptomics and proteomics. Apoptosis was selected for pathway-focused analysis due to its role in bacterial infection of tick cells. The results showed tissue-specific differences in tick response to infection and revealed differentiated regulation of apoptosis pathways. The impact of bacterial infection was more pronounced in tick nymphs and midguts than in salivary glands, probably reflecting bacterial developmental cycle. All apoptosis pathways described in other organisms were identified in I. scapularis, except for the absence of the Perforin ortholog. Functional characterization using RNA interference showed that Porin knockdown significantly increases tick colonization by A. phagocytophilum. Infection with A. phagocytophilum produced complex tissue-specific alterations in transcript and protein levels. In tick nymphs, the results suggested a possible effect of bacterial infection on the inhibition of tick immune response. In tick midguts, the results suggested that A. phagocytophilum infection inhibited cell apoptosis to facilitate and establish infection through up-regulation of the JAK/STAT pathway. Bacterial infection inhibited the intrinsic apoptosis pathway in tick salivary glands by down-regulating Porin expression that resulted in the inhibition of Cytochrome c release as the anti-apoptotic mechanism to facilitate bacterial infection. However, tick salivary glands may promote apoptosis to limit bacterial infection through induction of the extrinsic apoptosis pathway. These dynamic changes in response to A. phagocytophilum in I. scapularis tissue-specific transcriptome and proteome demonstrated the complexity of the tick response to infection and will contribute to characterize gene regulation in ticks.
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- 2015
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25. Studies of Anaplasma phagocytophilum in sheep experimentally infected with the human NY-18 isolate: characterization of tick feeding sites.
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Reppert E, Galindo RC, Ayllón N, Breshears MA, Kocan KM, Blouin EF, and de la Fuente J
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- Anaplasmosis microbiology, Animals, Ehrlichiosis microbiology, Female, Humans, Male, Models, Animal, Sheep, Sheep Diseases microbiology, Zoonoses, Anaplasma phagocytophilum physiology, Anaplasmosis transmission, Ehrlichiosis transmission, Host-Pathogen Interactions, Ixodes microbiology, Sheep Diseases transmission
- Abstract
Anaplasma phagocytophilum, transmitted by ticks of the genus Ixodes, was first described in Scotland as the agent of tick-borne fever in sheep and more recently as the cause of human granulocytic anaplasmosis in the U.S. and Europe. We previously reported sheep as an experimental host for the human NY-18 isolate of A. phagocytophilum. While clinical signs were not observed and infected granulocytes were not seen in stained blood smears, these sheep served as a good host for infection of ticks. In this research we characterized tick feeding sites to better understand tick/host/pathogen interactions. Ixodes scapularis adults were allowed to feed for 2 and 4 days on experimentally infected sheep, after which biopsies were taken beneath tick feeding sites for histopathology, PCR and immunohistochemistry (IHC) studies. In addition, the expression of selected immune response genes was studied in blood and feeding site biopsies. While necrosis was too advanced in 4-day biopsies for accurate cell counts, higher numbers of eosinophils and neutrophils were found in 2-day biopsies from infected sheep as compared with the uninfected controls. An unexpected result was the documentation of higher dermal inflammation in infected sheep at sites without ticks. A. phagocytophilum infected granulocytes were localized by immunohistochemistry (IHC) in skin biopsies using rabbit antibodies against the recombinant A. phagocytophilum major surface protein 4 as the primary antibody for indirect peroxidase-anti-peroxidase and fluorescent antibody IHC. These infected cells are likely to be the source of infection for ticks. Sheep therefore served as good hosts for studying host/pathogen/tick interactions of this human strain of A. phagocytophilum, and provided a means of producing infected ticks for future studies on tick/pathogen developmental and transmission cycles., (Copyright © 2014 Elsevier GmbH. All rights reserved.)
- Published
- 2014
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26. A systems biology approach to the characterization of stress response in Dermacentor reticulatus tick unfed larvae.
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Villar M, Popara M, Ayllón N, Fernández de Mera IG, Mateos-Hernández L, Galindo RC, Manrique M, Tobes R, and de la Fuente J
- Subjects
- Animals, Arachnid Vectors microbiology, Arthropod Proteins genetics, Arthropod Proteins metabolism, Dermacentor microbiology, Food Deprivation, Genes, Bacterial, Larva microbiology, Larva physiology, Protein Biosynthesis, Proteome genetics, Proteome metabolism, RNA, Messenger genetics, RNA, Messenger metabolism, Rickettsia genetics, Systems Biology, Transcriptome, Arachnid Vectors physiology, Dermacentor physiology, Stress, Physiological
- Abstract
Background: Dermacentor reticulatus (Fabricius, 1794) is distributed in Europe and Asia where it infests and transmits disease-causing pathogens to humans, pets and other domestic and wild animals. However, despite its role as a vector of emerging or re-emerging diseases, very little information is available on the genome, transcriptome and proteome of D. reticulatus. Tick larvae are the first developmental stage to infest hosts, acquire infection and transmit pathogens that are transovarially transmitted and are exposed to extremely stressing conditions. In this study, we used a systems biology approach to get an insight into the mechanisms active in D. reticulatus unfed larvae, with special emphasis on stress response., Principal Findings: The results support the use of paired end RNA sequencing and proteomics informed by transcriptomics (PIT) for the analysis of transcriptomics and proteomics data, particularly for organisms such as D. reticulatus with little sequence information available. The results showed that metabolic and cellular processes involved in protein synthesis were the most active in D. reticulatus unfed larvae, suggesting that ticks are very active during this life stage. The stress response was activated in D. reticulatus unfed larvae and a Rickettsia sp. similar to R. raoultii was identified in these ticks., Significance: The activation of stress responses in D. reticulatus unfed larvae likely counteracts the negative effect of temperature and other stress conditions such as Rickettsia infection and favors tick adaptation to environmental conditions to increase tick survival. These results show mechanisms that have evolved in D. reticulatus ticks to survive under stress conditions and suggest that these mechanisms are conserved across hard tick species. Targeting some of these proteins by vaccination may increase tick susceptibility to natural stress conditions, which in turn reduce tick survival and reproduction, thus reducing tick populations and vector capacity for tick-borne pathogens.
- Published
- 2014
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27. Interaction of the tick immune system with transmitted pathogens.
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Hajdušek O, Síma R, Ayllón N, Jalovecká M, Perner J, de la Fuente J, and Kopáček P
- Subjects
- Anaplasma pathogenicity, Animals, Arachnid Vectors microbiology, Arachnid Vectors parasitology, Babesia pathogenicity, Borrelia pathogenicity, Ticks microbiology, Ticks parasitology, Anaplasma immunology, Arachnid Vectors immunology, Babesia immunology, Borrelia immunology, Host-Pathogen Interactions, Immunity, Innate, Ticks immunology
- Abstract
Ticks are hematophagous arachnids transmitting a wide variety of pathogens including viruses, bacteria, and protozoans to their vertebrate hosts. The tick vector competence has to be intimately linked to the ability of transmitted pathogens to evade tick defense mechanisms encountered on their route through the tick body comprising midgut, hemolymph, salivary glands or ovaries. Tick innate immunity is, like in other invertebrates, based on an orchestrated action of humoral and cellular immune responses. The direct antimicrobial defense in ticks is accomplished by a variety of small molecules such as defensins, lysozymes or by tick-specific antimicrobial compounds such as microplusin/hebraein or 5.3-kDa family proteins. Phagocytosis of the invading microbes by tick hemocytes is likely mediated by the primordial complement-like system composed of thioester-containing proteins, fibrinogen-related lectins and convertase-like factors. Moreover, an important role in survival of the ingested microbes seems to be played by host proteins and redox balance maintenance in the tick midgut. Here, we summarize recent knowledge about the major components of tick immune system and focus on their interaction with the relevant tick-transmitted pathogens, represented by spirochetes (Borrelia), rickettsiae (Anaplasma), and protozoans (Babesia). Availability of the tick genomic database and feasibility of functional genomics based on RNA interference greatly contribute to the understanding of molecular and cellular interplay at the tick-pathogen interface and may provide new targets for blocking the transmission of tick pathogens.
- Published
- 2013
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28. Anaplasma phagocytophilum inhibits apoptosis and promotes cytoskeleton rearrangement for infection of tick cells.
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Ayllón N, Villar M, Busby AT, Kocan KM, Blouin EF, Bonzón-Kulichenko E, Galindo RC, Mangold AJ, Alberdi P, Pérez de la Lastra JM, Vázquez J, and de la Fuente J
- Subjects
- Anaplasma phagocytophilum genetics, Animals, Carrier Proteins genetics, Caspase 9 genetics, Caspase 9 metabolism, Cell Line, Feeding Behavior, Female, Gastrointestinal Tract microbiology, Gene Expression Regulation, Gene Knockdown Techniques, Host-Pathogen Interactions, Ixodes genetics, Ixodes metabolism, Male, Microfilament Proteins genetics, Mitochondrial Membrane Transport Proteins genetics, Mitochondrial Membrane Transport Proteins metabolism, Phylogeny, RNA Interference, RNA, Messenger genetics, RNA, Messenger metabolism, Salivary Glands microbiology, Spectrin genetics, Spectrin metabolism, Voltage-Dependent Anion Channels genetics, Voltage-Dependent Anion Channels metabolism, Anaplasma phagocytophilum pathogenicity, Apoptosis, Carrier Proteins metabolism, Cytoskeleton metabolism, Ixodes microbiology, Microfilament Proteins metabolism
- Abstract
Anaplasma phagocytophilum causes human granulocytic anaplasmosis. Infection with this zoonotic pathogen affects gene expression in both the vertebrate host and the tick vector, Ixodes scapularis. Here, we identified new genes, including spectrin alpha chain or alpha-fodrin (CG8) and voltage-dependent anion-selective channel or mitochondrial porin (T2), that are involved in A. phagocytophilum infection/multiplication and the tick cell response to infection. The pathogen downregulated the expression of CG8 in tick salivary glands and T2 in both the gut and salivary glands to inhibit apoptosis as a mechanism to subvert host cell defenses and increase infection. In the gut, the tick response to infection through CG8 upregulation was used by the pathogen to increase infection due to the cytoskeleton rearrangement that is required for pathogen infection. These results increase our understanding of the role of tick genes during A. phagocytophilum infection and multiplication and demonstrate that the pathogen uses similar strategies to establish infection in both vertebrate and invertebrate hosts.
- Published
- 2013
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29. Reciprocal regulation of NF-kB (Relish) and Subolesin in the tick vector, Ixodes scapularis.
- Author
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Naranjo V, Ayllón N, Pérez de la Lastra JM, Galindo RC, Kocan KM, Blouin EF, Mitra R, Alberdi P, Villar M, and de la Fuente J
- Subjects
- Animals, Antigens metabolism, Arthropod Proteins metabolism, Base Sequence, Conserved Sequence genetics, DNA Primers genetics, Electrophoresis, Capillary, Electrophoretic Mobility Shift Assay, Enzyme-Linked Immunosorbent Assay, Gene Components, Ixodes immunology, Models, Biological, Molecular Sequence Data, RNA Interference, Sequence Analysis, DNA, Antigens genetics, Arthropod Proteins genetics, Arthropod Vectors metabolism, Gene Expression Regulation immunology, Gene Regulatory Networks immunology, Ixodes metabolism, NF-kappa B metabolism
- Abstract
Background: Tick Subolesin and its ortholog in insects and vertebrates, Akirin, have been suggested to play a role in the immune response through regulation of nuclear factor-kappa B (NF-kB)-dependent and independent gene expression via interaction with intermediate proteins that interact with NF-kB and other regulatory proteins, bind DNA or remodel chromatin to regulate gene expression. The objective of this study was to characterize the structure and regulation of subolesin in Ixodes scapularis. I. scapularis is a vector of emerging pathogens such as Borrelia burgdorferi, Anaplasma phagocytophilum and Babesia microti that cause in humans Lyme disease, anaplasmosis and babesiosis, respectively. The genome of I. scapularis was recently sequenced, and this tick serves as a model organism for the study of vector-host-pathogen interactions. However, basic biological questions such as gene organization and regulation are largely unknown in ticks and other arthropod vectors., Principal Findings: The results presented here provide evidence that subolesin/akirin are evolutionarily conserved at several levels (primary sequence, gene organization and function), thus supporting their crucial biological function in metazoans. These results showed that NF-kB (Relish) is involved in the regulation of subolesin expression in ticks, suggesting that as in other organisms, different NF-kB integral subunits and/or unknown interacting proteins regulate the specificity of the NF-kB-mediated gene expression. These results suggested a regulatory network involving cross-regulation between NF-kB (Relish) and Subolesin and Subolesin auto-regulation with possible implications in tick immune response to bacterial infection., Significance: These results advance our understanding of gene organization and regulation in I. scapularis and have important implications for arthropod vectors genetics and immunology highlighting the possible role of NF-kB and Subolesin/Akirin in vector-pathogen interactions and for designing new strategies for the control of vector infestations and pathogen transmission.
- Published
- 2013
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30. Immunization with recombinant subolesin does not reduce tick infection with tick-borne encephalitis virus nor protect mice against disease.
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Havlíková S, Ličková M, Ayllón N, Roller L, Kazimírová M, Slovák M, Moreno-Cid JA, Pérez de la Lastra JM, Klempa B, and de la Fuente J
- Subjects
- Animals, Antigens metabolism, Arthropod Proteins metabolism, Down-Regulation, Encephalitis Viruses, Tick-Borne, Female, Immunization, Ixodes metabolism, Ixodes virology, Mice, Mice, Inbred BALB C, Recombinant Proteins immunology, Recombinant Proteins metabolism, Antigens immunology, Arthropod Proteins immunology, Encephalitis, Tick-Borne prevention & control, Ixodes immunology, Tick Infestations prevention & control, Viral Vaccines immunology
- Abstract
Tick-borne encephalitis (TBE) is a growing zoonotic disease caused by tick-borne encephalitis virus (TBEV) infection. Although effective vaccines for TBEV are available, on-going vaccination efforts are insufficient to prevent increase in TBE cases annually. Vaccination with arthropod vector antigens to reduce vector infestations and vector capacity allows control of several vector-borne diseases by targeting their common vector. Subolesin (SUB) is a tick protective antigen that has a role in tick innate immunity and other molecular pathways and has been shown to protect against tick infestations and infection by vector-borne pathogens. However, SUB expression and the effect of SUB immunization have not been evaluated for tick-borne viruses. Herein, we showed that SUB expression is downregulated during Ixodes ricinus tick feeding but induced in ticks infected with TBEV, thus supporting a role for this molecule in tick innate immune response to virus infection. Immunization with recombinant SUB reduced SUB mRNA levels in nymphs co-feeding with infected females and suggested and effect on tick infestations in mice. However, SUB immunization did not reduce tick infection with TBEV nor protect mice against TBE. These results suggested that SUB is not a good candidate antigen for vaccination against TBEV and support the characterization of tick-pathogen interactions to identify mechanisms that could be targeted to reduce TBEV infection and transmission by ticks., (Copyright © 2013 Elsevier Ltd. All rights reserved.)
- Published
- 2013
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31. Identification of microorganisms in partially fed female horn flies, Haematobia irritans.
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Torres L, Almazán C, Ayllón N, Galindo RC, Rosario-Cruz R, Quiroz-Romero H, Gortazar C, and de la Fuente J
- Subjects
- Animals, Cattle blood, Expressed Sequence Tags, Female, Mycobacterium bovis classification, Mycobacterium bovis genetics, Picornaviridae classification, Picornaviridae genetics, RNA-Directed DNA Polymerase, Real-Time Polymerase Chain Reaction, Wolbachia classification, Wolbachia genetics, Diptera microbiology, Diptera physiology, Mycobacterium bovis isolation & purification, Picornaviridae isolation & purification, Wolbachia isolation & purification
- Abstract
The horn fly Haematobia irritans (Linnaeus, 1758) (Diptera: Muscidae) is one of the most important ectoparasites of cattle. The parasitism of horn flies interferes with cattle feeding, thus reducing weight gain and milk production. Additionally, horn flies are mechanical vectors of pathogens that cause disease in cattle. The aims of this study were to identify microorganisms in partially fed female horn flies through mining of expressed sequence tags (ESTs) and to characterize microorganism prevalence using real-time RT-PCR. Seven unigenes containing 24 ESTs were homologous to infectious agents. Microorganisms identified in partially fed female horn flies ESTs included Nora virus (3 unigenes; 8 ESTs), Wolbachia endosymbionts (3 unigenes; 3 ESTs), and Mycobacterium bovis (1 unigene; 13 ESTs). These results expanded the repertoire of microorganisms that could cause persistent infections or be mechanically transmitted by horn flies and support further studies on the role of horn flies in the epidemiology of these pathogens in Mexico.
- Published
- 2012
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32. Sheep experimentally infected with a human isolate of Anaplasma phagocytophilum serve as a host for infection of Ixodes scapularis ticks.
- Author
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Kocan KM, Busby AT, Allison RW, Breshears MA, Coburn L, Galindo RC, Ayllón N, Blouin EF, and de la Fuente J
- Subjects
- Anaplasma phagocytophilum genetics, Anaplasma phagocytophilum immunology, Animals, Antigens, Bacterial immunology, Arachnid Vectors virology, Cell Line, DNA, Bacterial genetics, Ehrlichiosis complications, Enzyme-Linked Immunosorbent Assay, Female, Gastrointestinal Tract microbiology, Humans, Ixodes virology, Male, Models, Animal, Neutrophils microbiology, Nymph, Polymerase Chain Reaction, Salivary Glands microbiology, Sheep, Tick Infestations complications, Anaplasma phagocytophilum physiology, Arachnid Vectors microbiology, Ehrlichiosis microbiology, Ixodes microbiology, Tick Infestations parasitology
- Abstract
Anaplasma phagocytophilum, first identified as a pathogen of ruminants in Europe, has more recently been recognized as an emerging tick-borne pathogen of humans in the U.S. and Europe. A. phagocytophilum is transmitted by Ixodes spp., but the tick developmental cycle and pathogen/vector interactions have not been fully described. In this research, we report on the experimental infection of sheep with the human NY-18 isolate of A. phagocytophilum which then served as a host for infection of I. scapularis nymphs and adults. A. phagocytophilum was propagated in the human promyelocytic cell line, HL-60, and the infected cell cultures were then used to infect sheep by intravenous inoculation. Infections in sheep were confirmed by PCR and an Anaplasma-competitive ELISA. Clinical signs were not apparent in any of the infected sheep, and only limited hematologic and mild serum biochemical abnormalities were identified. While A. phagocytophilum morulae were rarely seen in neutrophils, blood film evaluation revealed prominent large granular lymphocytes, occasional plasma cells, and rare macrophages. Upon necropsy, gross lesions were restricted to the lymphoid system. Mild splenomegaly and lymphadenomegaly with microscopic evidence of lymphoid hyperplasia was observed in all infected sheep. Female I. scapularis that were allowed to feed and acquire infection on each of the 3 experimentally infected sheep became infected with A. phagocytophilum as determined by PCR of guts (80-87%) and salivary glands (67-100%). Female I. scapularis that acquired infection as nymphs on an experimentally infected sheep transmitted A. phagocytophilum to a susceptible sheep, thus confirming transstadial transmission. Sheep proved to be a good host for the production of I. scapularis infected with this human isolate of A. phagocytophilum, which can be used as a model for future studies of the tick/pathogen interface., (Copyright © 2012 Elsevier GmbH. All rights reserved.)
- Published
- 2012
- Full Text
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33. Characterization of the tick-pathogen interface by quantitative proteomics.
- Author
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Villar M, Popara M, Bonzón-Kulichenko E, Ayllón N, Vázquez J, and de la Fuente J
- Subjects
- Animals, Arachnid Vectors microbiology, Arthropod Proteins metabolism, Cells, Cultured, Humans, Tick Infestations metabolism, Tick Infestations parasitology, Ticks microbiology, Arachnid Vectors metabolism, Arthropod Proteins isolation & purification, Host-Pathogen Interactions physiology, Proteomics methods, Ticks metabolism
- Abstract
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., (Copyright © 2012 Elsevier GmbH. All rights reserved.)
- Published
- 2012
- Full Text
- View/download PDF
34. Impact of climate trends on tick-borne pathogen transmission.
- Author
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Estrada-Peña A, Ayllón N, and de la Fuente J
- Abstract
Recent advances in climate research together with a better understanding of tick-pathogen interactions, the distribution of ticks and the diagnosis of tick-borne pathogens raise questions about the impact of environmental factors on tick abundance and spread and the prevalence and transmission of tick-borne pathogens. While undoubtedly climate plays a role in the changes in distribution and seasonal abundance of ticks, it is always difficult to disentangle factors impacting on the abundance of tick hosts from those exerted by human habits. All together, climate, host abundance, and social factors may explain the upsurge of epidemics transmitted by ticks to humans. Herein we focused on tick-borne pathogens that affect humans with epidemic potential. Borrelia burgdorferi s.l. (Lyme disease), Anaplasma phagocytophilum (human granulocytic anaplasmosis), and tick-borne encephalitis virus (tick-borne encephalitis) are transmitted by Ixodes spp. Crimean-Congo hemorrhagic fever virus (Crimean-Congo hemorrhagic fever) is transmitted by Hyalomma spp. In this review, we discussed how vector tick species occupy the habitat as a function of different climatic factors, and how these factors impact on tick survival and seasonality. How molecular events at the tick-pathogen interface impact on pathogen transmission is also discussed. Results from statistically and biologically derived models are compared to show that while statistical models are able to outline basic information about tick distributions, biologically derived models are necessary to evaluate pathogen transmission rates and understand the effect of climatic variables and host abundance patterns on pathogen transmission. The results of these studies could be used to build early alert systems able to identify the main factors driving the subtle changes in tick distribution and seasonality and the prevalence of tick-borne pathogens.
- Published
- 2012
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35. Functional genomics of the horn fly, Haematobia irritans (Linnaeus, 1758).
- Author
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Torres L, Almazán C, Ayllón N, Galindo RC, Rosario-Cruz R, Quiroz-Romero H, and de la Fuente J
- Subjects
- Animals, Expressed Sequence Tags, Female, Gene Library, RNA Interference, Sequence Analysis, DNA, Genome, Insect, Genomics, Muscidae genetics
- Abstract
Background: The horn fly, Haematobia irritans (Linnaeus, 1758) (Diptera: Muscidae) is one of the most important ectoparasites of pastured cattle. Horn flies infestations reduce cattle weight gain and milk production. Additionally, horn flies are mechanical vectors of different pathogens that cause disease in cattle. The aim of this study was to conduct a functional genomics study in female horn flies using Expressed Sequence Tags (EST) analysis and RNA interference (RNAi)., Results: A cDNA library was made from whole abdominal tissues collected from partially fed adult female horn flies. High quality horn fly ESTs (2,160) were sequenced and assembled into 992 unigenes (178 contigs and 814 singlets) representing molecular functions such as serine proteases, cell metabolism, mitochondrial function, transcription and translation, transport, chromatin structure, vitellogenesis, cytoskeleton, DNA replication, cell response to stress and infection, cell proliferation and cell-cell interactions, intracellular trafficking and secretion, and development. Functional analyses were conducted using RNAi for the first time in horn flies. Gene knockdown by RNAi resulted in higher horn fly mortality (protease inhibitor functional group), reduced oviposition (vitellogenin, ferritin and vATPase groups) or both (immune response and 5'-NUC groups) when compared to controls. Silencing of ubiquitination ESTs did not affect horn fly mortality and oviposition while gene knockdown in the ferritin and vATPse functional groups reduced mortality when compared to controls., Conclusions: These results advanced the molecular characterization of this important ectoparasite and suggested candidate protective antigens for the development of vaccines for the control of horn fly infestations.
- Published
- 2011
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36. Characterization of pathogen-specific expression of host immune response genes in Anaplasma and Mycobacterium species infected ruminants.
- Author
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Galindo RC, Ayllón N, Carta T, Vicente J, Kocan KM, Gortazar C, and de la Fuente J
- Subjects
- Anaplasmosis genetics, Animals, Cattle, Cattle Diseases genetics, Gene Expression Profiling, Mycobacterium Infections genetics, Mycobacterium Infections immunology, Mycobacterium avium subsp. paratuberculosis immunology, Mycobacterium bovis immunology, Oligonucleotide Array Sequence Analysis, Polymerase Chain Reaction, Anaplasma ovis immunology, Anaplasmosis immunology, Cattle Diseases immunology, Deer genetics, Deer immunology, Deer microbiology, Gene Expression Regulation, Genes, MHC Class II, Mycobacterium immunology
- Abstract
Anaplasma and Mycobacterium species are among the most prevalent bacterial pathogens in European red deer (Cervus elaphus) in south-central Spain and are known to modify gene expression in ruminants. In this study, we used microarray hybridization and real-time RT-PCR analyses to characterize global gene expression profiles in red deer in response to Anaplasma ovis and A. ovis/Mycobacterium bovis/Mycobacterium avium sub. paratuberculosis (MAP) infections, compare the expression of immune response genes between red deer infected with A. ovis, M. bovis and A. ovis/M. bovis/MAP, and characterize the differential expression of immune response genes identified in red deer in cattle infected with M. bovis and Anaplasma marginale. Global gene differential expression in A. ovis- and A. ovis/M. bovis/MAP-infected deer resulted in the modification of common and pathogen-specific cellular biological processes. The differential expression of host immune response genes showed pathogen and host-specific signatures and the effect of infection with multiple pathogens on deer immune response. These results suggested that intracellular bacteria from Anaplasma and Mycobacterium genera produce similar genes expression patterns in infected ruminants. However, pathogen and host-specific differences could contribute to disease diagnosis and treatment in ruminants., (Copyright © 2010 Elsevier Ltd. All rights reserved.)
- Published
- 2010
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37. Expression of Heat Shock and Other Stress Response Proteins in Ticks and Cultured Tick Cells in Response to Anaplasma spp. Infection and Heat Shock.
- Author
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Villar M, Ayllón N, Busby AT, Galindo RC, Blouin EF, Kocan KM, Bonzón-Kulichenko E, Zivkovic Z, Almazán C, Torina A, Vázquez J, and de la Fuente J
- Abstract
Ticks are ectoparasites of animals and humans that serve as vectors of Anaplasma and other pathogens that affect humans and animals worldwide. Ticks and the pathogens that they transmit have coevolved molecular interactions involving genetic traits of both the tick and the pathogen that mediate their development and survival. In this paper, the expression of heat shock proteins (HSPs) and other stress response proteins (SRPs) was characterized in ticks and cultured tick cells by proteomics and transcriptomics analyses in response to Anaplasma spp. infection and heat shock. The results of these studies demonstrated that the stress response was activated in ticks and cultured tick cells after Anaplasma spp. infection and heat shock. However, in the natural vector-pathogen relationship, HSPs and other SRPs were not strongly activated, which likely resulted from tick-pathogen coevolution. These results also demonstrated pathogen- and tick-specific differences in the expression of HSPs and other SRPs in ticks and cultured tick cells infected with Anaplasma spp. and suggested the existence of post-transcriptional mechanisms induced by Anaplasma spp. to control tick response to infection. These results illustrated the complexity of the stress response in ticks and suggested a function for the HSPs and other SRPs during Anaplasma spp. infection.
- Published
- 2010
- Full Text
- View/download PDF
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