Person:
Domínguez Bernal, Gustavo Ramón

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First Name
Gustavo Ramón
Last Name
Domínguez Bernal
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Veterinaria
Department
Sanidad Animal
Area
Sanidad Animal
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Now showing 1 - 10 of 19
  • Publication
    Immunization 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 Ramón
    Leishmania 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.
  • Publication
    Strength 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 Ramón
    HisAK70candidateshave 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.
  • Publication
    Mitigating 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, Lara
    Leishmania 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.
  • Publication
    Mechanisms 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 La
    Several 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.
  • Publication
    Engineering 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 Ramón
    The 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.
  • Publication
    Characterisation of the ex vivo virulence of Leishmania infantum isolates from Phlebotomus perniciosus from an outbreak of human leishmaniosis in Madrid, Spain
    (BioMed Central Ltd., 2014-11-07) Jiménez, Maribel; Molina, Ricardo; Ordóñez-Gutiérrez, Lara; Carrión Herrero, Francisco Javier; Domínguez Bernal, Gustavo Ramón; Martínez Rodrigo, Abel; Mas Zubiri, Alicia; Cutuli Simón, María Teresa
    Background Since mid 2009, an outbreak of human leishmaniosis in Madrid, Spain, has involved more than 560 clinical cases. Many of the cases occurred in people who live in areas around a newly constructed green park (BosqueSur). This periurban park provides a suitable habitat for sand flies (the vectors of Leishmania infantum). Indeed, studies of blood meals from sand flies captured in the area showed a strong association between the insect vector, hares or rabbits, and humans in the area. Interestingly, up to 70% of cases have been found in immunocompetent patients (aged between 46-60 years). This study was designed to evaluate the ex vivo virulence of the L. infantum isolates from Phlebotomus perniciosus captured in this area of Madrid. Methods Murine macrophages and dendritic cells were infected ex vivo with L. infantum strain BCN150, isolate BOS1FL1, or isolate POL2FL7. At different times after infection, the infection indices, cytokine production (IL-12p40 and IL-10), NO release and arginase activities were evaluated. Results Using an ex vivo model of infection in murine bone marrow-derived cells, we found that infection with isolates BOS1FL1 and POL2FL7 undermined host immune defence mechanisms in multiple ways. The main factors identified were changes in both the balance of iNOS versus arginase activities and the equilibrium between the production of IL-12 and IL-10. Infection with isolates BOS1FL1 and POL2FL7 also resulted in higher infection rates compared to the BCN150 strain. Infection index values at 24 h were as follows: BCN150-infected cells, 110 for infected MØ and 115 for infected DC; BOS1FL1-infected cells, 300 for infected MØ and 247 for infected DC; and POL2FL7-infected cells, 275 for infected MØ and 292 for infected DC. Conclusions Our data indicate that L. infantum isolates captured from this endemic area exhibited high virulence in terms of infection index, cytokine production and enzymatic activities involved in the pathogenesis of visceral leishmaniosis. Altogether, these data provide a starting point for the study of the virulence behaviour of parasites (BOS1FL1 and POL2FL7) isolated from P. perniciosus during the outbreak of human leishmaniosis in Madrid, Spain, and their involvement in infecting immunocompetent hosts.
  • Publication
    Short communication: Isolation frequency of bacteria causing lymphadenitis and abscesses in small ruminants in central Spain
    (Elsevier, 2017-07-10) Heras, Moisés de las; Torrijos, Carlos; Carrión Herrero, Francisco Javier; Fuente López, Ricardo De La; Díez De Tejada Martín, María Paloma; Pérez Sancho, Marta; Orden Gutiérrez, José Antonio; Domínguez Bernal, Gustavo Ramón
    Infectious lymphadenitis in small ruminants caused by Corynebacterium pseudotuberculosis and Staphylococcus aureus subsp. anaerobius are widely distributed throughout the world, and result in significant economic losses. Trueperella pyogenes has also been associated with lymphadenitis in sheep and goats. In order to determinate the isolation frequency of the different agents associated with lymphadenitis and abscesses, 171 pus samples (135 from sheep and 36 from goats) from 46 flocks were investigated. Isolated bacteria were identified by MALDI-TOF method. S. aureus subsp. anaerobius was the most frequently detected agent. It was identified in 76 animals (59 sheep and 17 goats) from 24 of the surveyed flocks. Of these infected animal, 25 (32,9%) were over one year old, confirming that abscess disease may occur in a significant percentage of adult animals. C. pseudotuberculosis was identified in 45 of the sampled animals (36 sheep and 9 goats) from 24 flocks. Only 5 of animals suffering caseous lymphadenitis were under one year old. T. pyogenes was isolated from 17 animals (13 sheep and 4 goats) in 11 flocks. Seven of these samples were taken from subcutaneous abscesses located in not lymph nodes regions. A notable finding of this work is the isolation of Actinomyces hyovaginalis from 5 of the samples analyzed all of them taken from subcutaneous abscesses located in superficial lymph nodes regions. Thus, T. pyogenes and A. hyovaginalis should be included in the differential diagnosis of lymphadenitis in small ruminants. In 19 of the 46 surveyed flocks at least two of the four agents were detected, which underlines the need to analyze samples from several animals to reach an accurate diagnosis in flocks affected by lymphadenitis.
  • Publication
    Subtilase Cytotoxin-Coding Genes in Verotoxin-Producing Escherichia coli Strains from Sheep and Goats Differ from Those from Cattle
    (2011-09-24) Carrión Herrero, Francisco Javier; Orden Gutiérrez, José Antonio; Horcajo Iglesias, María Del Pilar; Fuente López, Ricardo De La; Ruiz Santa Quiteria Serrano De La Cruz, José Antonio; Domínguez Bernal, Gustavo Ramón
    Subtilase cytotoxin (SubAB) from verotoxin (VT)-producing Escherichia coli (VTEC) strains was first described in the 98NK2 strain and has been associated with human disease. However, SubAB has recently been found in two VT-negative E. coli strains (ED 591 and ED 32). SubAB is encoded by two closely linked, cotranscribed genes (subA and subB). In this study, we investigated the presence of subAB genes in 52 VTEC strains isolated from cattle and 209 strains from small ruminants, using PCR. Most (91.9%) VTEC strains from sheep and goats and 25% of the strains from healthy cattle possessed subAB genes. The presence of subAB in a high percentage of the VTEC strains from small ruminants might increase the pathogenicity of these strains for human beings. Some differences in the results of PCRs and in the association with some virulence genes suggested the existence of different variants of subAB. We therefore sequenced the subA gene in 12 strains and showed that the subA gene in most of the subAB-positive VTEC strains from cattle was almost identical (about 99%) to that in the 98NK2 strain, while the subA gene in most of the subAB-positive VTEC strains from small ruminants was almost identical to that in the ED 591 strain. We propose the terms subAB1 to describe the SubAB-coding genes resembling that in the 98NK2 strain and subAB2 to describe those resembling that in the ED 591 strain.
  • Publication
    Differences in virulence gene expression between atypical enteropathogenic Escherichia coli strains isolated from diarrheic and healthy ruminants
    (2012-06-07) Carrión Herrero, Francisco Javier; Horcajo Iglesias, María Del Pilar; Domínguez Bernal, Gustavo Ramón; Fuente López, Ricardo De La; Ruiz Santa Quiteria Serrano De La Cruz, José Antonio; Orden Gutiérrez, José Antonio
    Differences in the pathogenicity of atypical enteropathogenic Escherichia coli (EPEC) strains may be due, at least partially, to different expression patterns of some virulence genes. To investigate this hypothesis, the virulence gene expression patterns of 6 atypical EPEC strains isolated from healthy and diarrheic ruminants were compared using quantitative real-time reverse transcription polymerase chain reaction after growing the bacteria in culture medium alone or after binding it to HeLa epithelial cells. Some virulence genes in strains from diarrheic animals were upregulated relative to their expression in strains from healthy animals. When bacteria were cultured in the presence of HeLa cells, the ehxA and efa1/lifA genes, previously associated with the production of diarrhea, were expressed at higher levels in strains from diarrheic animals than in strains from healthy animals. Thus, the expression levels of some virulence genes may help determine which atypical EPEC strains cause diarrhea in ruminants.
  • Publication
    Ruminants are not a reservoir of enteroaggregative Escherichia coli
    (Universidad Austral de Chile, 2017) Ruiz-Santa-Quiteria, José A.; García, Ángel; Carrión Herrero, Francisco Javier; Orden Gutiérrez, José Antonio; Domínguez Bernal, Gustavo Ramón; Horcajo Iglesias, María Del Pilar
    Enteroaggregative Escherichia coli (EAEC) causes both acute and persistent diarrhoea among children and adults in developing and developed countries. In addition, the large outbreak of food-borne illness in Europe in the summer of 2011 was caused by a verotoxin-producing EAEC O104:H4 strain. The public health threat posed by EAEC makes it important to identify animal reservoirs that might lead to epidemics in humans. Based on only one study in sheep and a few in cattle, researchers have suggested that EAEC are not found in ruminants. To expand the available data on this question, the present study applied polymerase chain reaction to faecal samples from 920 cattle, sheep and goats to detect EAEC. None of the samples tested positive for any of the EAEC genes tested, suggesting that goats, like cattle and sheep, are unlikely to be a significant reservoir of EAEC.