Vacuna frente a infestaciones provocadas por artrópodos hematófagos

La presente invención describe una vacuna frente a infestaciones provocadas por artrópodos hematófagos preferentemente garrapatas, mosquitos, flebótomos, ácaros rojos de las gallinas, pulgas, piojos, piojos de mar, etc. Así mismo, se protege un péptido capaz de inducir una respuesta inmune protectora y de reacción cruzada frente a una infestación provocada por artrópodos hematófagos, un polinucleótido, un vector de expresión, una célula hospedadora, un animal no humano, un anticuerpo, y un medicamento, así como el uso de estos productos en medicina, preferentemente en tecnologías del ADN recombinante y en inmunologíaPeer reviewedConsejo Superior de Investigaciones Científicas, Universidad de Castilla La ManchaA1 Solicitud de patente con informe sobre el estado de la técnic

Vaccination with BM86, subolesin and akirin protective antigens for the control of tick infestations in white tailed deer and red deer

Red deer (Cervus elaphus) and white-tailed deer (Odocoileus virginianus) are hosts for different tick species and tick-borne pathogens and play a role in tick dispersal and maintenance in some regions. These factors stress the importance of controlling tick infestations in deer and several methods such as culling and acaricide treatment have been used. Tick vaccines are a cost-effective alternative for tick control that reduced cattle tick infestations and tick-borne pathogens prevalence while reducing the use of acaricides. Our hypothesis is that vaccination with vector protective antigens can be used for the control of tick infestations in deer. Herein, three experiments were conducted to characterize (1) the antibody response in red deer immunized with recombinant BM86, the antigen included in commercial tick vaccines, (2) the antibody response and control of cattle tick infestations in white-tailed deer immunized with recombinant BM86 or tick subolesin (SUB) and experimentally infested with Rhipicephalus (Boophilus) microplus, and (3) the antibody response and control of Hyalomma spp. and Rhipicephalus spp. field tick infestations in red deer immunized with mosquito akirin (AKR), the SUB ortholog and candidate protective antigen against different tick species and other ectoparasites. The results showed that deer produced an antibody response that correlated with the reduction in tick infestations and was similar to other hosts vaccinated previously with these antigens. The overall vaccine efficacy was similar between BM86 (E=76%) and SUB (E=83%) for the control of R. microplus infestations in white-tailed deer. The field trial in red deer showed a 25-33% (18-40% when only infested deer were considered) reduction in tick infestations, 14-20 weeks after the first immunization. These results demonstrated that vaccination with vector protective antigens could be used as an alternative method for the control of tick infestations in deer to reduce tick populations and dispersal in regions where deer are relevant hosts for these ectoparasites.This work was supported by the Ministerio de Ciencia e Innovación, Spain (project BFU2008-01244/BMC) and the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Spain (project FAU2008-00014-00-00). D. Carreón was partially supported by a grant-in-aid from Conacyt, Mexico and the EU grant TB-STEP 212414. M. Villar was funded by the JAE-DOC program (CSIC-FSE), Spain. J.A. Moreno-Cid is a recipient of a JCCM fellowship, Spain.Peer Reviewe

Reciprocal regulation of NF-kB (Relish) and subolesin in the tick vector, Ixodes scapularis

[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.This research was supported by grants BFU2008-01244/BMC and BFU2011-23896 to JF, the Oklahoma Agricultural Experimental Grant 1669 and the Walter R. Sitlington Endowed Chair for Food Animal Research to KMK, and the EU FP7 ANTIGONE project number 278976. V. Naranjo was funded by the European Social Fund and the Junta de Comunidades de Castilla-La Mancha (Program FSE 2007-2013), Spain. N. Ayllón was funded by Ministerio de Educación y Ciencia (MEC), Spain.Peer Reviewe

Reactivity and Applications of Singlet Oxygen Molecule

Reactive oxygen species (ROS) are molecules produced in living organisms, in the environment, and in various chemical reactions. The main species include, among others, singlet oxygen (1O2), the superoxide anion radical (•O2−), the hydroxyl radical (HO•), and the hydroperoxyl radical (HOO•). In general, the reactivity of 1O2 is lower than that of HO• but even higher than that of •O2−. Singlet oxygen is the lowest energy excited state of molecular oxygen, but it is also a highly reactive species, which can initiate oxidation reactions of biomolecules such as amino acids, proteins, nucleic acids, and lipids, either by a direct reaction or by the induction of ROS. Singlet oxygen is a highly reactive electrophilic species that reacts with electron-rich molecules and is related to several types of pathologies. To inhibit the oxidation of biomolecules with this species, some substances act as antioxidants by performing a quenching effect. In this chapter, aspects such as its physicochemical properties, methods of generation and detection, as well as the reactivity of this molecule are detailed

Bioinformatics Discovery of Vertebrate Cathelicidins from the Mining of Available Genomes

Due to the worrying increase in antimicrobial resistance to conventional antibiotics, the search for alternatives is becoming increasingly important. Antimicrobial peptides (AMPs), originating from natural resources, have been recognised as a novel class of antibiotics. An advantage of peptides over antibiotics is that the resistance is more difficult to attain than for conventional antibiotics. With the increasing number of genomes sequenced and available in the public domain, one alternative methodology to obtain novel AMPs is to analyse genes and proteins from genomic databases to predict and identify amino acid sequences that share similarities and molecular features with natural bioactive antimicrobial peptides. In this chapter, we summarise some of our recent results on the production of antimicrobial peptides, particularly, how we managed to identify a family of antimicrobial peptides: cathelicidins, through bioinformatics tools, from the genomes of two lower vertebrates (a reptile and a bird) available in public databases. We hope that our preliminary investigation with these novel peptides could be useful for the design of future strategies that pursue the production of antimicrobial peptides through biotechnology

No evidence that wild red deer (Cervus elaphus) on the Iberian Peninsula are a reservoir of Mycobacterium avium subspecies paratuberculosis infection

The potential role of red deer (Cervus elaphus) as a reservoir of Mycobacterium avium subspecies paratuberculosis (MAP) infection is largely unknown. A total of 332 wild red deer were investigated using post-mortem examination, bacteriology and serology. Only three animals (1.12%) were found to have lesions on histopathological examination and no MAP bacteria were recovered on culture. The results suggest it is unlikely that wild red deer make a significant contribution to the maintenance of MAP infection in the region. The cross-reactivity of the ELISAs used indicates this diagnostic modality is ineffective in the detection of MAP infection in this species. The implications of these results for the control of this important pathogen in both livestock and wildlife are discussed.This research was supported by MCINN Grants AGL2008-03875/FEDER and FP7EU/TB-STEP-212414.Peer Reviewe

Tonsils of the soft palate do not mediate the response of pigs to oral vaccination with heat-inactivated Mycobacterium bovis

Mycobacterium bovis causes animal tuberculosis (TB) in cattle, humans, and other mammalian species, including pigs. The goal of this study was to experimentally assess the responses of pigs with and without a history of tonsillectomy to oral vaccination with heat-inactivated M. bovis and challenge with a virulent M. bovis field strain, to compare pig and wild boar responses using the same vaccination model as previously used in the Eurasian wild boar (Sus scrofa), to evaluate the use of several enzyme-linked immunosorbent assays (ELISAs) and lateral flow tests for in vivo TB diagnosis in pigs, and to verify if these tests are influenced by oral vaccination with inactivated M. bovis. At necropsy, the lesion and culture scores were 20% to 43% higher in the controls than those in the vaccinated pigs. Massive M. bovis growth from thoracic tissue samples was observed in 4 out of 9 controls but in none of the 10 vaccinated pigs. No effect of the presence or absence of tonsils was observed on these scores, suggesting that tonsils are not involved in the protective response to this vaccine in pigs. The serum antibody levels increased significantly only after challenge. At necropsy, the estimated sensitivities of the ELISAs and dual path platform (DPP) assays ranged from 89% to 94%. In the oral mucosa, no differences in gene expression were observed in the control group between the pigs with and without tonsils. In the vaccinated group, the mRNA levels for chemokine (C-C motif) receptor 7 (CCR7), interferon beta (IFN-β), and methylmalonyl coenzyme A mutase (MUT) were higher in pigs with tonsils. Complement component 3 mRNA levels in peripheral blood mononuclear cells (PBMC) increased with vaccination and decreased after M. bovis challenge. This information is relevant for pig production in regions that are endemic for M. bovis and for TB vaccine research.This study is a contribution to EU FP7 grant 613779 WildTBVac and to MINECO Plan Nacional grant AGL2011-30041 and FEDER.Peer Reviewe

Genome editing technologies, mechanisms and improved production of therapeutic phytochemicals: Opportunities and prospects

Plants produce a large number of secondary metabolites, known as phytometabolites that may be employed as medicines, dyes, poisons, and insecticides in the field of medicine, agriculture, and industrial use, respectively. The rise of genome management approaches has promised a factual revolution in genetic engineering. Targeted genome editing in living entities permits the understanding of the biological systems very clearly, and also sanctions to address a wide-ranging objective in the direction of improving features of plant and their yields. The last few years have introduced a number of unique genome editing systems, including transcription activator-like effector nucleases, zinc finger nucleases, and miRNA-regulated clustered regularly interspaced short palindromic repeats/Cas9 (CRISPR/Cas9). Genome editing systems have helped in the transformation of metabolic engineering, allowing researchers to modify biosynthetic pathways of different secondary metabolites. Given the growing relevance of editing genomes in plant research, the exciting novel methods are briefly reviewed in this chapter. Also, this chapter highlights recent discoveries on the CRISPR-based modification of natural products in different medicinal plants.All the authors are highly grateful and acknowledge the authority ofthe respective departments and institutions for their support incarrying out this study. This research was funded by projectsAPOGEO (Cooperation Program INTERREG‐MAC 2014–2020, withEuropean Funds for Regional Development‐FEDER).“Agencia Canaria de Investigación, Innova‐ción y Sociedad de la Información (ACIISI)del Gobierno de Canarias”(Project ProID2020010134), and Fundación CajaCanarias (Project 2019SP43).Peer reviewe

Oral vaccination with heat inactivated Mycobacterium bovis activates the complement system to protect against tuberculosis

Tuberculosis (TB) remains a pandemic affecting billions of people worldwide, thus stressing the need for new vaccines. Defining the correlates of vaccine protection is essential to achieve this goal. In this study, we used the wild boar model for mycobacterial infection and TB to characterize the protective mechanisms elicited by a new heat inactivated Mycobacterium bovis vaccine (IV). Oral vaccination with the IV resulted in significantly lower culture and lesion scores, particularly in the thorax, suggesting that the IV might provide a novel vaccine for TB control with special impact on the prevention of pulmonary disease, which is one of the limitations of current vaccines. Oral vaccination with the IV induced an adaptive antibody response and activation of the innate immune response including the complement component C3 and inflammasome. Mycobacterial DNA/RNA was not involved in inflammasome activation but increased C3 production by a still unknown mechanism. The results also suggested a protective mechanism mediated by the activation of IFN-γ producing CD8+ T cells by MHC I antigen presenting dendritic cells (DCs) in response to vaccination with the IV, without a clear role for Th1 CD4+ T cells. These results support a role for DCs in triggering the immune response to the IV through a mechanism similar to the phagocyte response to PAMPs with a central role for C3 in protection against mycobacterial infection. Higher C3 levels may allow increased opsonophagocytosis and effective bacterial clearance, while interfering with CR3-mediated opsonic and nonopsonic phagocytosis of mycobacteria, a process that could be enhanced by specific antibodies against mycobacterial proteins induced by vaccination with the IV. These results suggest that the IV acts through novel mechanisms to protect against TB in wild boar.This research was supported by Plan Nacional I+D+I AGL2011-30041 from Ministerio de Economía y Competitividad (MINECO), Spain and FEDER. This is also a contribution to EU FP7 grant WildTBvac and the EU FP7 ANTIGONE project number 278976. R.C. Galindo was funded by MEC, Spain. B. Beltrán-Beck was supported by MINECO grant BES-2009-017401.Peer Reviewe