Person: Domínguez Bernal, Gustavo Ramón
Universidad Complutense de Madrid
Faculty / Institute
Now showing 1 - 10 of 26
PublicationVeterinaria es calidad: Evaluación contínua y autoevaluación(2018-03-22) Olmeda García, Angeles Sonia; Lorenzo González, Pedro Luis; Fernández Álvarez, Manuela; Cambero Rodríguez, María Isabel; Domínguez Bernal, Gustavo Ramón; Sainz Rodríguez, Ángel; San Andrés Larrea, María Dolores de; Pérez Cabal, María de los Ángeles; Pérez Sen, Raquel; Barrero Rodríguez, AndrésEl documento recoge los resultados del proyecto de Innova-Gestion UCM desarrollado durante los años 2016/2017 para la evaluación por rúbrica y online, formando, aplicando y analizando sus resultados. También se exponen los datos preliminares de la opinión de docentes y alumnos sobre su satisfacción con el actual Grado en Veterinaria. Estos resultados son, además, el punto de partida, para continuar mejorando la calidad de la docencia del centro. PublicationTranscriptomic Profile of Canine DH82 Macrophages Infected by Leishmania infantum Promastigotes with Different Virulence Behavior(MPDI, 2022-01-27) Mas, Alicia; Martínez Rodrigo, Abel; Carrión, Javier; Orden, José Antonio; Alzate, Juan F.; Domínguez Bernal, Gustavo Ramón; Horcajo, PilarZoonotic visceral leishmaniosis caused by Leishmania infantum is an endemic disease in the Mediterranean Basin affecting mainly humans and dogs, the main reservoir. The leishmaniosis outbreak declared in the Community of Madrid (Spain) led to a significant increase in human disease incidence without enhancing canine leishmaniosis prevalence, suggesting a better adaptation of the outbreak’s isolates by other host species. One of the isolates obtained in the focus, IPER/ES/2012/BOS1FL1 (BOS1FL1), has previously demonstrated a different phenotype than the reference strain MCAN/ES/1996/BCN150 (BCN150), characterized by a lower infectivity when interacting with canine macrophages. Nevertheless, not enough changes in the cell defensive response were found to support their different behavior. Thus, we decided to investigate the molecular mechanisms involved in the interaction of both parasites with DH82 canine macrophages by studying their transcriptomic profiles developed after infection using RNA sequencing. The results showed a common regulation induced by both parasites in the phosphoinositide-3-kinase–protein kinase B/Akt and NOD-like receptor signaling pathways. However, other pathways, such as phagocytosis and signal transduction, including tumor necrosis factor, mitogen-activated kinases and nuclear factor-κB, were only regulated after infection with BOS1FL1. These differences could contribute to the reduced infection ability of the outbreak isolates in canine cells. Our results open a new avenue to investigate the true role of adaptation of L. infantum isolates in their interaction with their different hosts. PublicationImmunization with the HisAK70 DNA Vaccine Induces Resistance against Leishmania Amazonensis Infection in BALB/c Mice(MDPI, 2019-11-14) Martínez Rodrigo, Abel; S. Dias, Daniel; Ribeiro, Patrícia A. F.; Roatt, Bruno M.; Mas Zubiri, Alicia; Carrión Herrero, Francisco Javier; Coelho, Eduardo A. F.; Domínguez Bernal, Gustavo RamónLeishmania amazonensis is the aetiological agent of a broad spectrum of leishmaniosis in South America. It can cause not only numerous cases of cutaneous leishmaniosis but also diffuse cutaneous leishmaniosis. Considering the diversity of parasite species causing different forms of the disease that coexist in the same region, it is desirable to develop a vaccine capable of eliciting cross-protection. We have previously described the use of HisAK70 DNA vaccine for immunization of mice to assess the induction of a resistant phenotype against Leishmania major and infantum infections. In this study, we extended its application in the murine model of infection by using L. amazonensis promastigotes. Our data revealed that 14 weeks post-infection, HisAK70-vaccinated mice showed key biomarkers of protection, such as higher iNOS/arginase activity, IFN-γ/IL-10, IFN-γ/IL-4, and GM-CSF/IL-10 ratios, in addition to an IgG2a-type response when compared to the control group. These findings correlated with the presentation of lower footpad swelling and parasite burdens in the immunized compared to the control mice. Overall, this study suggests that immunization with HisAK70 may be considered a suitable tool to combat leishmaniosis as it is able to induce a potent cellular immune response, which allows to control the infection caused by L. amazonensis. PublicationStrength and medium-term impact of HisAK70 immunization in dogs: Vaccine safety and biomarkers of effectiveness for ex vivo Leishmania infantum infection(Elsevier, 2019-08) Fernández-Cotrina; Javier; Belinchón-Lorenzo, Silvia; Arias, Pablo; Martínez Rodrigo, Abel; Mas Zubiri, Alicia; Orden Gutiérrez, José Antonio; Fuente López, Ricardo De La; Carrión Herrero, Francisco Javier; Domínguez Bernal, Gustavo RamónHisAK70candidateshave successfullybeentested incutaneous (CL) andvisceral leishmaniosis (VL)mouse models.Here,weanalysedifferentbiomarkersindogtrialsafteraheterologousimmunizationstrategywitha HisAK70candidate(plasmidDNAplusadoptivetransferofperipheralblood-deriveddendriticcells(DCs)pulsed withthesamepathoantigenandCpGODNasanadjuvant)toexploretheantileishmanialactivityinanexvivo canineco-culturesysteminthepresenceofLeishmaniainfantumparasites.Inthecaninemodel,theheterologous HisAK70vaccinecoulddecreasetheinfectionindexintheDC-Tcellco-culturesystembyupto54%after30days andreachalmost67%after100dayspost-immunization,respectively,comparedtothoseobtainedinthecontrol groupofdogs.Theobservedsecurityandpotential tofight exvivoL. infantuminfectionhighlightaHisAK70 heterologousimmunizationstrategyasapromisingalternativetoevaluateitseffectivenessagainstcanineVL. PublicationDetection and antimicrobial resistance of Enterobacteriaceae other than Escherichia coli in raccoons from the Madrid region of Spain(SCIENDO, 2022-11-04) Orden Gutiérrez, José Antonio; Martínez Rodrigo, Abel; Vela Alonso, Ana Isabel; Fernández-Garayzábal Fernández, José Francisco; Hurtado Morillas, Clara; Mas Zubiri, Alicia; Domínguez Bernal, Gustavo RamónRaccoons are an invasive alien species widely distributed in the Madrid region of Spain. These animals can carry a variety of enteric bacteria with associated antimicrobial resistance, which can infect humans and livestock. However, to our knowledge, the presence of non-E. coli Enterobacteriaceae in raccoons has not been previously studied. We conducted a study to examine the species distribution of Enterobacteriaceae isolates other than E. coli, as well as their antimicrobial resistance, in the faeces of 83 raccoons in the Madrid region. We detected 12 Enterobacteriaceae isolates other than E. coli belonging to seven different species: Citrobacter freundii (1 isolate), Citrobacter gillenii (3 isolates), Citrobacter murliniae (1 isolate), Citrobacter portucalensis (2 isolates), Enterobacter hormaechei subsp. hoffmannii (1 isolate), Hafnia paralvei (2 isolates) and Raoultella ornithinolytica (2 isolates). These isolates were found in 7 of the 83 (8.4%) animals studied. To our knowledge, this study is the first report of the presence of non-E. coli Enterobacteriaceae in raccoon faeces. All isolates but one were resistant to at least one of the 14 antimicrobials tested. Resistance to ampicillin (83.3%), amoxicillinclavulanic acid (50%) and cefoxitin (33.3%) was the most frequent. Our study indicates that raccoons are a potential source of infection with Enterobacteriaceae other than E. coli for humans and livestock in the Madrid region. PublicationA further investigation of the leishmaniosis outbreak in Madrid (Spain): low-infectivity phenotype of the Leishmania infantum BOS1FL1 isolate to establish infection in canine cells(Elsevier, 2020-12) Viñals, Luis Miguel; Mas Zubiri, Alicia; Martínez Rodrigo, Abel; Orden Gutiérrez, José Antonio; Domínguez Bernal, Gustavo Ramón; Carrión Herrero, Francisco JavierHuman leishmaniosis caused by Leishmania infantum is a zoonotic disease, with dogs as the main reservoir in Mediterranean Basin countries. The largest European outbreak of human leishmaniosis declared in the southwestern Madrid region (Spain) is characterized by unusual epidemiological and clinical features, such as the emergence of new wild reservoirs (hares and rabbits), whereas the seroprevalence, infection, and severity of canine leishmaniosis have not substantially changed since the first studies conducted in Madrid before the outbreak. Previous studies reported that L. infantum isolates from the Madrid leishmaniosis focus displayed elevated virulence in in vivo models of infection and increased infectivity in murine target cells. With the aim of studying whether changes in the host-parasite interaction and virulence profile have developed, we first assessed the behaviour of one circulating isolate of the outbreak, IPER/ES/2012/BOS1FL1 (BOS1FL1), compared to that of a well-characterized strain from canine leishmaniosis, MCAN/ES/1996/BCN150 (BCN150), in terms of infection capacity (percentage of infected cells, representing infectivity, and number of amastigotes per infected cell, representing the intensity of infection) in canine monocytes and macrophages. BCN150 displayed significantly higher infectivity (76.82 ±4.40 vs 38.58 ±2.19; P <0.0001) and intensity of infection (3.64 ±0.13 vs 1.83 ±0.12; P <0.0001) than BOS1FL1 when interacting with canine cells. Our ROS induction results did not differ significantly between the two isolates or with the responses previously described for other L. infantum isolates. Paradoxically, increased resilience to hydrogen peroxide exposure was observed for BOS1FL1 (% viability 40.62 ±5.54 vs 26.37 ±2.93; P =0.039). Finally, we demonstrated that a decreased intracellular load of BOS1FL1 was associated with increased IFN-γ (261.21 ±26.29 vs 69.80 ±9.02; P =0.0151) and decreased IL- 10 production (165.06 ±23.87 vs 264.41 ±30.58; P =0.0002). In this study, we provide the first detailed insight into the differences between the isolate BOS1FL1 from the outbreak in Madrid and the well-characterized strain BCN150 MON-1 obtained from a dog in their response to interacting with canine cells. However, further studies are necessary to shed light on the immune mechanisms resulting in BOS1FL1 exhibiting less virulent behaviour in canine cells than in cells derived from other host species. PublicationEpitope Selection for Fighting Visceral Leishmaniosis: Not All Peptides Function the Same Way(MDPI, 2020-07-01) Álvarez-Campos, Daniel; Martínez Rodrigo, Abel; Mas Zubiri, Alicia; Orden Gutiérrez, José Antonio; Domínguez Bernal, Gustavo Ramón; Carrión Herrero, Francisco JavierVisceral leishmaniosis (VL) caused by Leishmania infantum is a disease with an increasing prevalence worldwide. Treatments are expensive, toxic, and ineffective. Therefore, vaccination seems to be a promising approach to control VL. Peptide-based vaccination is a useful method due to its stability, absence of local side effects, and ease of scaling up. In this context, bioinformatics seems to facilitate the use of peptides, as this analysis can predict high binding affinity epitopes to MHC class I and II molecules of different species. We have recently reported the use of HisAK70 DNA immunization in mice to induce a resistant phenotype against L. major, L. infantum, and L. amazonensis infections. In the present study, we used bioinformatics tools to select promising multiepitope peptides (HisDTC and AK) from the polyprotein encoded in the HisAK70 DNA to evaluate their immunogenicity in the murine model of VL by L. infantum. Our results revealed that both multiepitope peptides were able to induce the control of VL in mice. Furthermore, HisDTC was able to induce a better cell-mediated immune response in terms of reduced parasite burden, protective cytokine profile, leishmanicidal enzyme modulation, and specific IgG2a isotype production in immunized mice, before and after infectious challenge. Overall, this study indicates that the HisDTC chimera may be considered a satisfactory tool to control VL because it is able to activate a potent CD4+ and CD8+ T-cell protective immune responses. PublicationMitigating an undesirable immune response of inherent susceptibility to cutaneous leishmaniosis in a mouse model: the role of the pathoantigenic HISA70 DNA vaccine(2012-08-09) Herrero-Gil, Aldara; Carrión Herrero, Francisco Javier; Domínguez Bernal, Gustavo Ramón; Horcajo Iglesias, María Del Pilar; Orden Gutiérrez, José Antonio; Fuente López, Ricardo De La; Ordóñez Gutiérrez, LaraLeishmania major is the major cause of cutaneous leishmaniosis (CL) outside of the Americas. In the present study we have cloned six Leishmania genes (H2A, H2B, H3, H4, A2 and HSP70) into the eukaryotic expression vector pCMVβ-m2a, resulting in pCMV-HISA70m2A, which encodes all six pathoantigenic proteins as a single polyprotein. This expression plasmid has been evaluated as a novel vaccine candidate in the BALB/c mouse model of CL. The DNA vaccine shifted the immune response normally induced by L. major infection away from a Th2-specific pathway to one of basal susceptibility. Immunization with pCMV-HISA70m2A dramatically reduced footpad lesions and lymph node parasite burdens relative to infected control mice. Complete absence of visceral parasite burden was observed in all 12 immunized animals but not in any of the 24 control mice. Moreover, vaccinated mice produced large amounts of IFN-γ, IL-17 and NO at 7 weeks post-infection (pi), and they showed lower arginase activity at the site of infection, lower IL-4 production and a weaker humoral immune response than infected control mice. Taken together, these results demonstrate the ability of the HISA70 vaccine to shift the murine immune response to L. major infection away from an undesirable, Th2-specific pathway to a less susceptible-like pathway involving Th1 and Th17 cytokine profiles. PublicationMechanisms of resistance and susceptibility to experimental visceral leishmaniosis: BALB/c mouse versus syrian hamster model(2011-02-23) Nieto, Ana; Madrid-Elena, Nadia; Carrión Herrero, Francisco Javier; Domínguez Bernal, Gustavo Ramón; Orden Gutiérrez, José Antonio; Fuente López, Ricardo De LaSeveral animal models have been established to study visceral leishmaniosis (VL), a worldwide vector-borne disease affecting humans and domestic animals that constitutes a serious public health problem. BALB/c mice and Syrian hamsters are the most widely used experimental models. In this paper, we summarize the advantages and disadvantages of these two experimental models and discuss the results obtained using these models in different studies of VL. Studies using the BALB/c mouse model have underscored differences between the liver and spleen in the course of VL, indicating that pathological evaluation of the visceral organs is essential for understanding the immune mechanisms induced by Leishmania infantum infection. The main goal of this review is to collate the relevant literature on Leishmania pathogenesis into a sequence of events, providing a schematic view of the main components of adaptive and innate immunity in the liver and spleen after experimental infection with L. infantum or L. donovani. This review also presents several viewpoints and reflections about some controversial aspects of Leishmania research, including the choice of experimental model, route of administration, inoculum size and the relevance of pathology (intimately linked to parasite persistence): a thorough understanding of which is essential for future VL research and the successful development of efficient control strategies for Leishmania spp. PublicationEngineering of a live Salmonella enterica serovar Choleraesuis negative-marker strain that allows serological differentiation between immunised and infected animals(Elsevier, 2016-07-01) Herrero-Gil, Aldara; Carrión Herrero, Francisco Javier; Orden Gutiérrez, José Antonio; Fuente López, Ricardo De La; Domínguez Bernal, Gustavo RamónThe usefulness of Salmonella vaccine vehicles is limited by the fact that control programmes relying on Salmonella bacteriology and serology cannot differentiate infected animals from vaccinated ones, an ability referred to as DIVA (differentiating infected from vaccinated animals). As a first step towards Salmonella-based DIVA vaccines, the ompA gene was deleted in live attenuated ΔphoP and ΔrpoS vaccine strains. The ompA gene is present in all Salmonella enterica serovars and it encodes an abundant, highly immunogenic outer membrane protein. The double mutant ΔphoP ΔompA and ΔrpoS ΔompA strains showed similar virulence attenuation, safety and immunogenicity in a mouse model of infection as the parental ΔphoP and ΔrpoS strains. Sera from mice inoculated with the double mutant strains failed to recognise OmpA in Western blots of outer membrane extracts, whereas the protein was recognised by sera from mice inoculated with wild-type Salmonella or a mixture of double mutant and parental strains. These data suggest that OmpA can be a suitable negative marker for DIVA vaccines.