Maria Montoya

• Autonomous University of Barcelona

Extracellular vesicles (EVs) released by virus-infected cells have the potential to encapsulate viral peptides, a characteristic that could facilitate vaccine development. Furthermore, plasma-derived EVs may elucidate pathological changes occurring in distal tissues during viral infections. We hypothesized that molecular characterization of EVs iso...

Due to the emergence of antibiotic resistance and new and more complex diseases that affect livestock animal health and food security, the control of epidemics has become a top priority worldwide. Vaccination represents the most important and cost-effective measure to control infectious diseases in animal health, but it represents only 23% of the t...

Dendritic cells (DCs) are the most potent antigen presenting cells (APCs). Because of the difficulty in obtaining these cells directly from tissues, different sources of DCs are frequently used for in vitro experimentation and many of their biological and functional characteristics were studied using these systems. Until recently, it was assumed th...

Despite the role of pigs as a source of new Influenza A Virus viruses (IAV) potentially capable of initiating human pandemics, immune responses to swine influenza virus (SwIV) in pigs are not fully understood. Several SwIV epitopes presented by swine MHC (SLA) class I have been identified using different approaches either in outbred pigs or in Babr...

Oseltamivir is a common therapy against influenza A virus (IAV) infections. The acquisition of oseltamivir resistance (OR) mutations, such as H275Y, hampers viral fitness. However, OR H1N1 viruses have demonstrated the ability to spread throughout different populations. The objective of this work was to compare the fitness of two strains of OR (R6...

Following short immunization protocols, naturally attenuated African swine fever virus (ASFV) isolate OURT88/3 and deletion mutant BeninΔMGF have previously been shown to induce high percentage of protection in domestic pigs against challenge with virulent virus. Results obtained in the present study showed that a single intramuscular immunization...

Classical approaches to African swine fever virus (ASFV) vaccine development have not been successful; inactivated virus does not provide protection and use of live attenuated viruses generated by passage in tissue culture had a poor safety profile. Current African swine fever (ASF) vaccine research focuses on the development of modified live virus...

Supplementary tables S1 to S3

Porcine respiratory disease complex (PRDC) is a polymicrobial syndrome that results from a combination of infectious agents, such as environmental stressors, population size, management strategies, age, and genetics. PRDC results in reduced performance as well as increased mortality rates and production costs in the pig industry worldwide. This rev...

The porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important swine diseases in the world. It is causing an enormous economic burden due to reproductive failure in sows and a complex respiratory syndrome in pigs of all ages, with mortality varying from 2 to 100% in the most extreme cases of emergent highly pathogenic...

The Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is the etiological agent of one of the most important swine diseases with a significant economic burden worldwide. Unfortunately, available vaccines are partially effective highlighting the need of novel approaches. Previously, antigenic viral proteins were described in serum-derived e...

Background The interaction between influenza virus and the host response to infection clearly plays an important role in determining the outcome of infection. While much is known on the participation of inflammation on the pathogenesis of severe A (H1N1) pandemic 09-influenza virus, its role in the course of non-fatal pneumonia has not been fully a...

Variation of gene expression along time in the infected group. Highlighted genes in colour represent genes involved in the top 20 canonical pathways identified by IPA. Orange: represent cytokines or chemokines genes and blue represent interferon stimulated genes. Red represents granzyme molecules.

List of genes differentially expressed between infected mice and controls. FC: fold change. Highlighted genes in colour represent genes involved in the top 20 canonical pathways identified by IPA. Orange: represent cytokines or chemokines genes and blue represent interferon stimulated genes. Red represents granzyme molecules.

Genes involved in the top 20 canonical signaling pathways altered by A (H1N1) pdm 09 virus at day 5 post infection. FC: fold change.

Droplet digital PCR validation of microarray data. Expression values obtained from the microarrays for IFNB1 and IL6 genes showed a significant positive correlation, confirmed by using digital droplet PCR.

Pulmonary gene expression profiles at day 5 and 10 post infection. A) Venn diagram showing those genes whose expression levels differed from controls either at day 5 and day 10, and those which differed only at one time point. B) Heatmap of the common signature across different time points. The colour is proportional to their fold change (FC) compa...

Studies evaluating host transcriptomic responses to A (H1N1) pdm09 influenza virus in animal models. This table summarized the most significant previous studies evaluating the transcriptomic response to A (H1N1) pdm09 virus in different animal models. EID50: 50% Egg Infective Dose, TCID50: 50% Tissue Culture Infective Dose, PFU: Plaque Forming Unit...

The current circulating swine influenza virus (IV) subtypes in Europe (H1N1, H1N2, and H3N2) are associated with clinical outbreaks of disease. However, we showed that pigs could be susceptible to other IV strains that are able to cross the species barrier. In this work, we extended our investigations into whether different IV strains able to cross...

Introduction Human and mouse respiratory tracts show anatomical and physiological differences, making it necessary to develop alternative experimental models for many respiratory diseases study. Pig has been recognized as a valuable biomedical model, in particular for lung transplantation or pathologies such as cystic fibrosis and influenza infecti...

[This corrects the article on p. 131 in vol. 4, PMID: 27900319.].

PRRSV is the etiological agent of one of the most important swine diseases with a significant economic burden worldwide and limitations in vaccinology. Exosomes are 30-100 nm vesicles of endocytic origin. Remarkably, immunizations with exosomes containing antigens from tumors or pathogens are capable of eliciting protective immune responses, albeit...

Severe cases after pH1N1 infection are consequence of interstitial pneumonia triggered by alveolar viral replication and an exacerbated host immune response, characterized by the up-regulation of pro-inflammatory cytokines and the influx of inflammatory leukocytes to the lungs. Different lung cell populations have been suggested as culprits in the...

PRRSV is the etiological agent of one of the most important swine diseases with a significant economic burden worldwide and limitations in vaccinology. Exosomes are 30–100 nm vesicles of endocytic origin. Remarkably, immunizations with exosomes containing antigens from tumors or pathogens are capable of eliciting protective immune responses, albeit...

Reticulocyte-derived exosomes (rex) are 30–100 nm membrane vesicles of endocytic origin released during the maturation of reticulocytes to erythrocytes upon fusion of multivesicular bodies with the plasma membrane. Combination of CpG-ODN with rex obtained from BALB/c mice infected with the reticulocyte-prone non-lethal P. yoelii 17X malaria strain...

List of mouse proteins detected by MS/MS.

Over-represented Gene ontology (GO) categories in the core proteome of mouse rexPy.

Gating strategy to define human splenocyte populations. Splenocytes were labeled with a panel of antibodies in order to discriminate immune relevant splenocyte populations (CD3-PE, CD4-APC, CD8-APC-H7, CD45-PerCP, CD16-V450, CD19-Bv510, CD56-PE-Bio770) and analyzed in a BD LSRFortessa flow cytometer. (A) Singlet cells were gated according to FSC-A/...

List of Plasmodium yoelii proteins detected by MS/MS.

Over-represented Gene ontology (GO) categories in the core proteome of Plasmodium yoelii from mouse rexPy.

Virus-like particles (VLPs), comprised of viral structural proteins devoid of genetic material, are tunable nanoparticles that can be chemically or genetically engineered, to be used as platforms for multimeric display of foreign antigens. Here, we report the engineering of chimeric VLPs, derived from rabbit hemorrhagic disease virus (RHDV) for pre...

Exosomes represent a new and advantageous approach to be evaluated in PRRSV, being that to the best of our knowledge is the first time that exosomes derived from PRRSV naïve and non-viremic swine has been characterized. Moreover, its immunological properties in the infectious diseases context makes them interesting as platforms for vaccination and...

Human and mouse respiratory tracts show anatomical and physiological differences, which will benefit from alternative experimental models for studying many respiratory diseases. Pig has been recognized as a valuable biomedical model, particularly for lung transplantation or pathologies such as cystic fibrosis and influenza infection. However, there...

Importance section: Although natural infection of humans with an avian H3N8 influenza A virus has not yet been reported, this influenza A virus subtype has already crossed the species barrier. Therefore, we have examined the potential of H3N8 from canine, equine, avian and seal origin to productively infect pigs. Our results demonstrated that avia...

It is well established that trans-placental transmission of classical swine fever virus (CSFV) during mid-gestation can lead to persistently infected offspring. The aim of the present study was to evaluate the ability of CSFV to induce viral persistence upon early postnatal infection. Two litters of 10 piglets each were infected intranasally on the...

The majority of pandemic 2009 H1N1 (A(H1N1)pdm09) influenza virus (IV) caused mild symptoms in most infected patients, however, a greater rate of severe disease was observed in healthy young adults and children without co-morbid conditions. The purpose of this work was to study in ferrets the dynamics of infection of two contemporary strains of hum...

GenBank Accession Numbers KP636483-KP636490 represent sequences from the 8 segments of Influenza A virus (A/Anas platyrhynchos/Spain/1877/2009(H7N2)). ##Assembly-Data-START## Assembly Method :: Lasergene v. 11 Sequencing Technology :: Illumina ##Assembly-Data-END##

GenBank Accession Numbers KP636483-KP636490 represent sequences from the 8 segments of Influenza A virus (A/Anas platyrhynchos/Spain/1877/2009(H7N2)). ##Assembly-Data-START## Assembly Method :: Lasergene v. 11 Sequencing Technology :: Illumina ##Assembly-Data-END##

GenBank Accession Numbers KP636483-KP636490 represent sequences from the 8 segments of Influenza A virus (A/Anas platyrhynchos/Spain/1877/2009(H7N2)). ##Assembly-Data-START## Assembly Method :: Lasergene v. 11 Sequencing Technology :: Illumina ##Assembly-Data-END##

GenBank Accession Numbers KP636483-KP636490 represent sequences from the 8 segments of Influenza A virus (A/Anas platyrhynchos/Spain/1877/2009(H7N2)). ##Assembly-Data-START## Assembly Method :: Lasergene v. 11 Sequencing Technology :: Illumina ##Assembly-Data-END##

GenBank Accession Numbers KP636483-KP636490 represent sequences from the 8 segments of Influenza A virus (A/Anas platyrhynchos/Spain/1877/2009(H7N2)). ##Assembly-Data-START## Assembly Method :: Lasergene v. 11 Sequencing Technology :: Illumina ##Assembly-Data-END##

GenBank Accession Numbers KP636483-KP636490 represent sequences from the 8 segments of Influenza A virus (A/Anas platyrhynchos/Spain/1877/2009(H7N2)). ##Assembly-Data-START## Assembly Method :: Lasergene v. 11 Sequencing Technology :: Illumina ##Assembly-Data-END##

GenBank Accession Numbers KP636483-KP636490 represent sequences from the 8 segments of Influenza A virus (A/Anas platyrhynchos/Spain/1877/2009(H7N2)). ##Assembly-Data-START## Assembly Method :: Lasergene v. 11 Sequencing Technology :: Illumina ##Assembly-Data-END##

GenBank Accession Numbers KP636483-KP636490 represent sequences from the 8 segments of Influenza A virus (A/Anas platyrhynchos/Spain/1877/2009(H7N2)). ##Assembly-Data-START## Assembly Method :: Lasergene v. 11 Sequencing Technology :: Illumina ##Assembly-Data-END##

Long pentraxin 3 (PTX3) is a conserved pattern-recognition secreted protein and a host-defence-related component of the humoral innate immune system. The aim of the present study was to characterize swine PTX3 (SwPTX3) protein expression in influenza virus infected pigs. First, we performed in silico studies to evaluate the cross-reactivity of PTX3...

Vaccination is considered one of the most effective ways to control pathogens and prevent diseases in humans as well as in the veterinary field. Traditional vaccines against animal viral diseases are based on inactivated or attenuated viruses, but new subunit vaccines are gaining attention from researchers in animal vaccinology. Among these, virus-...

in press

Profile of cytokines induction on A549 infected cells at 24 hpi. Cultured cells were infected at 3 PFU/cell with the A/New Caledonia/20/99 (NC) and the H1N1 pandemic M and F isolates. At 24 hpi cell supernatants were taken and used to determine the concentration of the indicated cytokines using the Luminex 100 System. Error bars indicate the standa...

Frequency of different residues at PB2 221, PA 529 and HA 127 positions in human, swine and avian viruses. (PDF)

Ccr5 allele determination. Samples of cultured A549 (top) and kidneys from infected or uninfected mice (bottom)were used to isolate total RNA that was used for RT-PCR reactions and DNA sequencing to detect the CCR5 region that comprises the 32-bp deletion. The sequence of the CCR5 allele from human (CCR5 H. sapiens) or mice (Ccr5 M. musculus) have...

Nucleotides at the indicated positions of M and F RNAs. (PDF)

Swine influenza virus is one of the most important pathogens involved in the swine respiratory disease complex. Recent serological surveys showed a high prevalence of swine influenza strains belonging to the H1N1, H1N2 and H3N2 subtypes circulating in pigs in Spain. However, little is known about their genome sequence. Five swine influenza strains...

Fluorescence images of co-infected-poBMDCs with H3N2 SwIV and SW114 or Nagasaki at 8 hpi. Porcine BMDC were infected and stained using (a) anti-SW14 or (b) anti-Nagasaki rabbit serum, and then with the anti-rabbit IgG-Dye Light 549, and anti-mouse NP HB65 ATCC antibody followed by an anti-mouse IgG-FITC. Therefore, nuclei were stained with DAPI (bl...

TEM of poBMDC co-infected with H3N2 SwIV and Nagasaki at 8 h. Porcine BMDC were infected with SwIV followed by Nagasaki. At 8 hpi, poBMDC showed several vesicles (ves) (a-e). Few cells showed drastic cell damage (b). Some vesicles contained more than one bacterium (d, g). The Golgi of Nagasaki-infected poBMDC was dilated (f, h). Different levels of...

H3N2 SwIV and H. parasuis SW114 or Nagasaki co-infected-poBMDC at (a) 1h and (b)8 hpi. Porcine BMDC were infected and stained using anti-SW14 or anti-Nagasaki rabbit serum 1 h at 4°C, and then with the anti-rabbit IgG-FITC antibody. Mock (grey histograms), SW114 (dotted line), Nagasaki (continuous line). Representative results of two independent ex...

TEM of poBMDC co-infected with H3N2 SwIV and SW114 at 8 h. Porcine BMDC showed many vesicles (ves) containing SW114 (a,b,c). Some vesicles had more than one bacteria (d, e). SW114 was found inside vesicles at different levels of degradation (asterisk, e, f, g). Also virus-like-particles (VLP) were observed (h, i). Bars: a = 2 μm; b-g = 200 nm; h =...

Vaccination continues to be the main approach to protect animals from infectious diseases. Until recently, all licensed vaccines were developed using conventional technologies. Subunit vaccines are, however, gaining attention from researchers in the field of veterinary vaccinology, and among these, virus-like particles (VLPs) represent one of the m...

The ability to control an immune response for the benefit and production efficiency of animals is the objective of immunomodulation in food-producing animals; substances that exert this control are called immunomodulators. A Spanish product (Inmunicín MAYMO®), based on food plant phytosterols, is being commercialized as complementary feed. The main...

Swine influenza virus (SwIV) causes sub-acute or acute respiratory infections on swine farms and pigs can act as "mixing vessels" for new influenza strains. Knowledge of the immune response of SwIV in its natural host, pigs, is very limited. Dendritic cells (DCs) mediate the induction of immunity to pathogens, but their interaction with SwIV has no...

ABSTRACT: The present study examined the immunological response of antigen presenting cells (APC) to genotype-I isolates of porcine reproductive and respiratory syndrome virus (PRRSV) infection by analysing the cytokine profile induced and evaluating the changes taking place upon infection on immunologically relevant cell markers (MHCI, MHCII, CD80...

The recent pandemic caused by human influenza virus A(H1N1) 2009 contains ancestral gene segments from North American and Eurasian swine lineages as well as from avian and human influenza lineages. The emergence of this A(H1N1) 2009 poses a potential global threat for human health and the fact that it can infect other species, like pigs, favours a...

The average of fragment kinetic energy (E*) and the multiplicity of prompt neutrons (nu) as a function of fragment mass (m*), as well as the fragment mass yield (Y(m*)) from thermal neutron-induced fission of 239Pu have been measured by Tsuchiya et al.. In that work the mass and kinetic energy are calculated from the measured kinetic energy of one...

Porcine circovirus type 2 (PCV2) is the essential etiological agent of postweaning multisystemic wasting syndrome (PMWS), a worldwide distributed pig disease. The involvement of the immune system in the pathogenesis of PMWS is considered crucial. Previous studies have shown a cytokine profile suggesting T immunosuppression and indicating that inter...

The aim of this study was to characterize the immune responses of DCs after infection with four different EU strains of PRRSV and whether they show any ability to immunomodulate T cells activation. Our results show that all EU strains can efficiently infect and replicate in DCs. Nevertheless, SLA-II levels remained unaltered in DC infected by all E...

Classical swine fever (CSF) is a highly contagious viral infection affecting domestic and wild pigs. For classical swine fever virus (CSFV), immunization with plasmids expressing different versions of glycoprotein E2 has proven an effective way to induce protection. Previously, we have also shown that immunization with DNA vaccine expressing glycop...

Type 1 IFNs, innate cytokines with important effector and immunomodulatory properties, are rapidly induced in the acute phase of many virus infections; however, this is generally a transient response that is not sustained during virus persistence. To gain insight into mechanisms that can contribute to down-regulation of type 1 IFN production during...

We have analyzed the potential of virus-like particles (VLPs) from rabbit hemorrhagic disease virus (RHDV) as a delivery system for foreign T cell epitopes. To accomplish this goal, we generated chimeric RHDV-VLPs incorporating a CD8(+) T cell epitope (SIINFEKL) derived from chicken ovalbumin (OVA). The OVA epitope was inserted in the capsid protei...

We investigated the contribution of lipopolysaccharide (LPS) to adjuvant properties of native outer membrane vesicles (NOMV), a vaccine candidate for meningococcal B disease. NOMV induce the maturation of and cytokine production by murine bone marrow-derived dendritic cells through both toll-like receptors (TLR) 2 and 4 which are mostly dependent o...

Las mediciones sobre la desviación estándar de la distribución de energía cinetica final (σe) en función de la masa final (m) de los fragmentos de la fisión de baja energía del 234U, hechas por Belhafaf et al., presentan un pico alrededor de m = 109 y otro alrededor de m = 122. Los autores atribuyen el primer pico a la evaporación de un elevado núm...

Porcine circovirus type 2 (PCV2) is a single-stranded circular DNA virus infecting domestic pigs worldwide. Interaction of this virus with the immune system apparently modulates the immune response of the host. In the present study, the implication of different components of PCV2 in the modulation of the immune response of the host were investigate...

Porcine circovirus type 2 (PCV2) is the necessary agent for the occurrence of post-weaning multisystemic wasting syndrome (PMWS) in pigs. It has been suggested that PMWS-affected pigs are immunosuppressed and, therefore, more prone to develop co-infections. In this study, we elucidated that PCV2 downregulates in vitro the immune cell functions duri...

The potential of PRRSv‐infected dendritic cells (DCs) to induce Foxp3+CD25+ T‐regulatory cells was evaluated in vitro using two American and five European strains. Monocyte‐derived DCs were infected, cultured for 24 h and in some cultures were treated with IFN‐α. Infected DCs were co‐cultured with autologous non‐adherent cells for 5 days and some c...

Immunomodulation in PRRS-MLV vaccinated pigs

Inmunomodulación del virus vacunal PRRSV

A recent report has shown that different PRRSV strains may have different abilities to induce neutralizing antibodies and, if produced, cross reactivity with other strains may be variable1. Moreover, other reports suggested that cell-mediated immunity, measured as IFN-γ-SC, may also have a role in PRRSV clearance2,3. However, cross reactivity of di...

In previous studies, we observed that different European-type strains may induce different cytokine patterns in antigen-presenting cells1. These different patterns might influence the outcome of the infection and have a role in the development of immunity. In PRRSV immunology is assumed that infection with a given strain provides long-term homologo...

The mass and kinetic energy distribution of nuclear fragments from the thermal neutron-induced fission of 235U have been studied using a Monte Carlo simulation. Besides reproducing the pronounced broadening on the standard deviation of the final fragment kinetic energy distribution (¾e(m)) around the mass number m = 109, our simulation also produce...

Modulation of the immunity during a PRRSV modified-live vaccination

Current vaccines for PRRSV are far from being universally efficacious. One of the main reasons for that is thought to be the high genetic diversity of PRRSV. In addition, it has been shown that protection induced by a given vaccine against a heterologous strain may be related to the ability of a given vaccine to induce strong cell-mediated response...

CD45 is a leukocyte tyrosine phosphatase, essential for normal immune responses. We have studied the function of splenic dendritic cells of CD45(+/+), CD45(-/-), CD45RABC and CD45RO transgenic mice. We show that there are increased numbers of plasmacytoid dendritic cells in CD45(-/-) mice. DC of all mice are capable of responding to lymphocytic cho...

Chimeric mice generated with bone marrow from RelB-deficient (-/-), RelB-heterozygous (+/-) and wild-type (+/+) mice were used to determine how total or partial absence of the transcription factor RelB in haematopoietic cells affects the immune response generated after lymphocytic choriomeningitis virus (LCMV) infection. In RelB(-/-) chimeras, earl...

Human coinfection with the helminth parasite Schistosoma mansoni and hepatitis B and hepatitis C viruses is associated with increased hepatic viral burdens and severe liver pathology. In this study we developed a murine S. mansoni/lymphocytic choriomeningitis virus (LCMV) coinfection model that reproduces the enhanced viral replication and liver pa...

Spain is the country with the highest rate of donation from deceased individuals. However, given the deficit in organs, living donation is being encouraged. Our objective was to analyze attitudes toward living donation among patients on the waiting list for a transplant. Patients on the waiting list for a kidney or liver transplant (n = 96; 46 kidn...