Victoria ley | Universidad Autónoma de Madrid (original) (raw)
Papers by Victoria ley
Res Immunol, 1990
The induction of specific cytotoxic T lymphocytes (CTL) is one component in the immune response w... more The induction of specific cytotoxic T lymphocytes (CTL) is one component in the immune response which can effectively protect the host against the progression of many viral infections. CTL are also known to play an important role in immune defence against tumour growth. CTL induction is dependent the presence of the specific antigen, appropriately presented, and interleukin-2 (IL2), provided by T helper lymphocytes. We studied the specific CTL response induced by tumour cells transfected with murine IL2. Our results show that tumour cells manipulated to secrete IL2 induce an improved specific anti-tumour response which results in tumour rejection in mice. To further investigate the effect of IL2 on the CTL response_ to_ different_.. _. .. _~nti~, ,n~.., . . . . . , u,,~..,. introduced synthetic peptides into IL2-secreting tumour cells and determined the specific CTL induction in syngeneic mice immunized with these cells. We report here that such IL2-secreting cells can effectively prime peptide-specific CTL in vivo. Our data are relevant to immunotherapy and vaccine develop° ment and open up the possibility that autologou s cells, manipulated to secrete IL2 and located with one or a cocktail of peptides, could be used to stimulate a specific CTL response.
RIV Rassegna Italiana di Valutazione, 2013
Animal Health Research Reviews
Swine vesicular disease is a highly contagious disease of pigs that is caused by an enterovirus o... more Swine vesicular disease is a highly contagious disease of pigs that is caused by an enterovirus of the family Picornaviridae. The virus is a relatively recent derivative of the human coxsackievirus B5, with which it has high molecular and antigenic homology. The disease is not severe, and affected animals usually show moderate general weakening and slight weight loss that is recovered in few days, as well as vesicular lesions in the mucosa of the mouth and nose and in the interdigital spaces of the feet. However, the similarity of these lesions to those caused by foot-and-mouth disease virus has led to the inclusion of this virus in list A of the Office International des Epizooties. The disease has been eradicated in the European Union except in Italy, where it is considered endemic in the south. Nevertheless, as occasional outbreaks still appear and must be eliminated rapidly, European countries are on the alert and farms are monitored routinely for the presence of the virus. This ...
La enfermedad vesicular del cerdo es una enfermedad altamente contagiosa que afecta al ganado por... more La enfermedad vesicular del cerdo es una enfermedad altamente contagiosa que afecta al ganado porcino, causada por un enterovirus de la familia Picornaviridae. El virus es un derivado relativamente reciente del virus humano coxsackie-B5, con el que tiene gran homología molecular y antigénica. La enfermedad no es grave y los animales se suelen recuperar en pocos días. Sin embargo, suelen aparecer vesículas en el hocico y en los espacios interdigitales de las pezuñas. El hecho de que estos síntomas sean indistinguibles de los causados por el virus de la fiebre aftosa, hace que este virus esté incluido dentro de la lista A de la Oficina Internacional de Epizootias. Actualmente la enfermedad está erradicada en los países de la Unión Europea excepto en Italia, donde se considera endémica. Sin embargo, en Europa se mantiene el estado de alarma por la posible aparición de brotes, ya que esporádicamente aparecen en algunos países y han de ser erradicados inmediatamente. Para ello se ha invertido un considerable esfuerzo en investigación de métodos de diagnóstico eficaces y en el estudio del virus y la patología de la enfermedad, información que resumimos en esta revisión. PALABRAS CLAVE: Enfermedad vesicular EVC Virus Enterovirus Cerdo Diagnóstico Patología Respuesta inmune
Research in immunology
The induction of specific cytotoxic T lymphocytes (CTL) is one component in the immune response w... more The induction of specific cytotoxic T lymphocytes (CTL) is one component in the immune response which can effectively protect the host against the progression of many viral infections. CTL are also known to play an important role in immune defence against tumour growth. CTL induction is dependent the presence of the specific antigen, appropriately presented, and interleukin-2 (IL2), provided by T helper lymphocytes. We studied the specific CTL response induced by tumour cells transfected with murine IL2. Our results show that tumour cells manipulated to secrete IL2 induce an improved specific anti-tumour response which results in tumour rejection in mice. To further investigate the effect of IL2 on the CTL response to different antigens, we introduced synthetic peptides into IL2-secreting tumour cells and determined the specific CTL induction in syngeneic mice immunized with these cells. We report here that such IL2-secreting cells can effectively prime peptide-specific CTL in vivo....
Acta crystallographica. Section D, Biological crystallography, 2003
Three different crystal forms of the swine vesicular disease virus (SVDV), isolate SPA/2/'93,... more Three different crystal forms of the swine vesicular disease virus (SVDV), isolate SPA/2/'93, were obtained by the hanging-drop vapour-diffusion technique using ammonium sulfate and sodium/potassium phosphate as precipitants. Monoclinic crystals, space group C2, with unit-cell parameters a = 473.7, b = 385.3, c = 472.8 A, beta = 100.4 degrees, contain one virus pArticle in the crystal asymmetric unit and diffract to 3.0 A resolution. A second type of crystals had a cubic morphology and diffracted beyond 2.6 A resolution. These crystals belong to a primitive orthorhombic space group, with unit-cell parameters a = 319.6, b = 353.8, c = 377.7 A, and contain half a virus pArticle in the asymmetric unit. A third type of crystals, with a prismatic shape and belonging to space group I222, was also obtained under similar crystallization conditions. These latter crystals, with unit-cell parameters a = 318.3, b = 349.9, c = 371.7 A, diffract to at least 3.0 A resolution and contain 15 pro...
Developments in biological standardization, 1999
Foot-and-mouth disease virus (FMDV) has been one of the pioneering viral systems in the developme... more Foot-and-mouth disease virus (FMDV) has been one of the pioneering viral systems in the development of synthetic peptides as vaccines. Protection against FMDV infection is associated with the induction of neutralising antibodies. Therefore, attempts have been made to identify peptides capable of eliciting protective humoral responses. Peptides based on a continuous, immunodominant B cell site on the capsid protein VP1 have been shown to confer limited protection in natural hosts. This probably reflects the difficulties in reproducing the immunogenicity of an entire viral particle by using a much simpler synthetic antigen, due to: (i) the polymorphism of the class II MHC; (ii) the adequate presentation to the immune system of the peptides, and (iii) the difficulties of achieving protection against a highly variable RNA virus, which may favour selection of virus antigenic variants. The improvement of FMD peptide vaccines, and the development of in vitro alternatives to in vivo immunog...
The Journal of general virology, 1999
A recombinant live vector vaccine was produced by insertion of cDNA encoding the structural prote... more A recombinant live vector vaccine was produced by insertion of cDNA encoding the structural proteins (P1) of foot-and-mouth disease virus (FMDV) into a replication-competent human adenovirus type 5 vaccine strain (Ad5 wt). Groups of cattle (n = 3) were immunized twice, by the subcutaneous and/or intranasal routes, with either the Ad5 wt vaccine or with the recombinant FMDV Ad5-P1 vaccine. All animals were challenged by intranasal instillation of FMDV 4 weeks after the second immunizations. In the absence of a detectable antibody response to FMDV, significant protection against viral challenge was seen in all of the animals immunized twice by the subcutaneous route with the recombinant vaccine. The observed partial protection against clinical disease was not associated with a reduction in titre of persistent FMDV infections in the oropharynx of challenged cattle.
Tesis doctoral inédita de la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento d... more Tesis doctoral inédita de la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 20-10-1997 Bibliografía:h. 107-121
La enfermedad vesicular del cerdo es una enfermedad altamente contagiosa que afecta al ganado por... more La enfermedad vesicular del cerdo es una enfermedad altamente contagiosa que afecta al ganado porcino, causada por un enterovirus de la familia Picornaviridae. El virus es un derivado relativamente reciente del virus humano coxsackie-B5, con el que tiene gran homología molecular y antigénica. La enfermedad no es grave y los animales se suelen recuperar en pocos días. Sin embargo, suelen aparecer vesículas en el hocico y en los espacios interdigitales de las pezuñas. El hecho de que estos síntomas sean indistinguibles de los causados por el virus de la fiebre aftosa, hace que este virus esté incluido dentro de la lista A de la Oficina Internacional de Epizootias. Actualmente la enfermedad está erradicada en los países de la Unión Europea excepto en Italia, donde se considera endémica. Sin embargo, en Europa se mantiene el estado de alarma por la posible aparición de brotes, ya que esporádicamente aparecen en algunos países y han de ser erradicados inmediatamente. Para ello se ha invertido un considerable esfuerzo en investigación de métodos de diagnóstico eficaces y en el estudio del virus y la patología de la enfermedad, información que resumimos en esta revisión. PALABRAS CLAVE: Enfermedad vesicular EVC Virus Enterovirus Cerdo Diagnóstico Patología Respuesta inmune
Methods in Biotechnology, 2006
Page 1. 153 From: Methods in Biotechnology, Vol. 21: Food-Borne Pathogens: Methods and Protocols ... more Page 1. 153 From: Methods in Biotechnology, Vol. 21: Food-Borne Pathogens: Methods and Protocols Edited by: CC Adley © Humana Press Inc., Totowa, NJ 13 Detection of Enteroviruses Miguel-Angel Jiménez-Clavero, Victoria Ley, Nuria Gómez, and Juan-Carlos Sáiz ...
Virology, 2000
Swine vesicular disease virus (SVDV) is an enterovirus of the Picornaviridae family that belongs ... more Swine vesicular disease virus (SVDV) is an enterovirus of the Picornaviridae family that belongs to the coxsackievirus B group. A number of antigenic sites have been identified in SVDV by analysis of neutralizing monoclonal antibody-resistant mutants and shown to be exposed on the surface of the capsid. In this paper we have identified seven new immunodominant antigenic regions in SVDV capsid proteins by a peptide scanning method, using a panel of sera from infected pigs. When these antigenic regions were located in the capsid by using a computer-generated three-dimensional model of the virion, one was readily exposed on the surface of the virus and the remaining sites were located facing the inner side of the capsid shell, at subunit contacts, or in the interior of the subunit structure.
Virology, 1999
The importance of the induction of virus neutralizing antibodies to provide protection against fo... more The importance of the induction of virus neutralizing antibodies to provide protection against foot-and-mouth disease virus (FMDV) infection is well established. However, recent studies with recombinant adenovirus expressing the precursor polypeptide of the viral capsid (P1) indicate that cattle inoculated with this recombinant vector developed partial protection against FMDV infection, in the absence of a detectable specific humoral response. Other viral vectors have been widely used to induce protective immunity against many pathogens, and it has been reported that the use of different vectors for priming and boosting injections can provide a synergistic effect on this response. In this work, we determined the immunogenicity of two recombinant viruses (adenovirus and vaccinia) expressing P1-FMDV, administered either individually or sequentially, and the protection that they induced against FMDV challenge in pigs. A double immunization with the adeno-P1 virus was the most effective strategy at inducing protective immunity. In contrast to previous reports, the use of two different vectors for priming and boosting did not show a synergistic effect on the protection induced against FMD. Interestingly, immunized pigs developed FMDV-specific T cell responses but not detectable antibodies. Thus, the protection observed was likely to be mediated by a cellular immune response.
Virology, 1984
of African swine fever virus DNA (about 170 kbp) with the restriction endonucleases SolI, EcoRI, ... more of African swine fever virus DNA (about 170 kbp) with the restriction endonucleases SolI, EcoRI, Kpn1, PvuI, and SmaI yielded 14,31,17,13, and 11 fragments, respectively. The order of the restriction fragments produced by each nuclease was established by identifying the crosslinked EcoRI and Sal1 terminal fragments and then finding overlapping fragments. The five restriction fragment maps were integrated into a single map by locating SalI, KpnI, PvuI, and SvzaI sites in cloned EcoRI fragments, and orienting each fragment in the overall map.
Virology, 1984
African swine fever virus DNA (about 170 kbp) was cleaved with the restriction endonuclease EwRI ... more African swine fever virus DNA (about 170 kbp) was cleaved with the restriction endonuclease EwRI and most of the resulting 31 fragments were cloned in either the phage vector hWES.hB or the plasmid pBR325. Three fragments were not cloned in those vectors, the largest fragment EcoRI-A (21.2 kbp) and the two crosslinked terminal fragments, EcoRI-K' and D'. Endonuclease Sal1 cut fragment EcoRI-A into three pieces which were cloned in plasmid pBR322. The two terminal EcoRI fragments were cloned after removal of the crosslinks with nuclease Sl and addition of EcoRI linkers to the fragment ends. The complete library of the cloned fragments accounted for about 98% of ASF virus genome, the missing sequences being those removed by the nuclease Sl in the process of cloning the terminal fragments.
Molecular Immunology, 1986
Mouse monoclonal antibodies (MAbs) with different specificities against Dermatophagoides farinae ... more Mouse monoclonal antibodies (MAbs) with different specificities against Dermatophagoides farinae (D. farinae) extract have been obtained. Fifteen of these antibodies reacted with allergen molecules contained in D. farinae and D. pteronyssinus extracts, immunoprecipitating the main allergen of D. farinae (DF29) and homologous allergen of D. pteronyssinus (DP28). In addition, the monoclonal antibody MADF2 immunoprecipitated DF29 together with two other polypeptides (mol. wt 20,000 and 40,000) from D. farinae extracts. Five monoclonal antibodies (MADF2, MADF5, MADF9, MADF10 and MADF13) were selected to study their epitope specificity and the relationship of the epitope location on the allergen with the IgE binding site. By cross-inhibition studies two different epitopes and two partly overlapping determinants were found. In addition, two of these epitopes, those defined by MADF13 and MADF5, are close to, or overlapping, IgE binding site(s) on the allergen molecule. DF29 allergen from D. farinae extract was purified by affinity chromatography using MADF5 coupled to Sepharose. The purified allergen had capacity to bind mite specific human IgE and demonstrated an allergenic activity of up to 70% of total extract of D. farinae. These results indicate that DF29 molecule is the main allergen from D. farinae extracts.
Molecular Immunology, 1985
Allergen molecules from Purietaria judaica pollen, a widely distributed allergy inducer in Southe... more Allergen molecules from Purietaria judaica pollen, a widely distributed allergy inducer in Southern and Western Europe, have been studied using specific monoclonal antibodies (MAbs). MAbs
Res Immunol, 1990
The induction of specific cytotoxic T lymphocytes (CTL) is one component in the immune response w... more The induction of specific cytotoxic T lymphocytes (CTL) is one component in the immune response which can effectively protect the host against the progression of many viral infections. CTL are also known to play an important role in immune defence against tumour growth. CTL induction is dependent the presence of the specific antigen, appropriately presented, and interleukin-2 (IL2), provided by T helper lymphocytes. We studied the specific CTL response induced by tumour cells transfected with murine IL2. Our results show that tumour cells manipulated to secrete IL2 induce an improved specific anti-tumour response which results in tumour rejection in mice. To further investigate the effect of IL2 on the CTL response_ to_ different_.. _. .. _~nti~, ,n~.., . . . . . , u,,~..,. introduced synthetic peptides into IL2-secreting tumour cells and determined the specific CTL induction in syngeneic mice immunized with these cells. We report here that such IL2-secreting cells can effectively prime peptide-specific CTL in vivo. Our data are relevant to immunotherapy and vaccine develop° ment and open up the possibility that autologou s cells, manipulated to secrete IL2 and located with one or a cocktail of peptides, could be used to stimulate a specific CTL response.
RIV Rassegna Italiana di Valutazione, 2013
Animal Health Research Reviews
Swine vesicular disease is a highly contagious disease of pigs that is caused by an enterovirus o... more Swine vesicular disease is a highly contagious disease of pigs that is caused by an enterovirus of the family Picornaviridae. The virus is a relatively recent derivative of the human coxsackievirus B5, with which it has high molecular and antigenic homology. The disease is not severe, and affected animals usually show moderate general weakening and slight weight loss that is recovered in few days, as well as vesicular lesions in the mucosa of the mouth and nose and in the interdigital spaces of the feet. However, the similarity of these lesions to those caused by foot-and-mouth disease virus has led to the inclusion of this virus in list A of the Office International des Epizooties. The disease has been eradicated in the European Union except in Italy, where it is considered endemic in the south. Nevertheless, as occasional outbreaks still appear and must be eliminated rapidly, European countries are on the alert and farms are monitored routinely for the presence of the virus. This ...
La enfermedad vesicular del cerdo es una enfermedad altamente contagiosa que afecta al ganado por... more La enfermedad vesicular del cerdo es una enfermedad altamente contagiosa que afecta al ganado porcino, causada por un enterovirus de la familia Picornaviridae. El virus es un derivado relativamente reciente del virus humano coxsackie-B5, con el que tiene gran homología molecular y antigénica. La enfermedad no es grave y los animales se suelen recuperar en pocos días. Sin embargo, suelen aparecer vesículas en el hocico y en los espacios interdigitales de las pezuñas. El hecho de que estos síntomas sean indistinguibles de los causados por el virus de la fiebre aftosa, hace que este virus esté incluido dentro de la lista A de la Oficina Internacional de Epizootias. Actualmente la enfermedad está erradicada en los países de la Unión Europea excepto en Italia, donde se considera endémica. Sin embargo, en Europa se mantiene el estado de alarma por la posible aparición de brotes, ya que esporádicamente aparecen en algunos países y han de ser erradicados inmediatamente. Para ello se ha invertido un considerable esfuerzo en investigación de métodos de diagnóstico eficaces y en el estudio del virus y la patología de la enfermedad, información que resumimos en esta revisión. PALABRAS CLAVE: Enfermedad vesicular EVC Virus Enterovirus Cerdo Diagnóstico Patología Respuesta inmune
Research in immunology
The induction of specific cytotoxic T lymphocytes (CTL) is one component in the immune response w... more The induction of specific cytotoxic T lymphocytes (CTL) is one component in the immune response which can effectively protect the host against the progression of many viral infections. CTL are also known to play an important role in immune defence against tumour growth. CTL induction is dependent the presence of the specific antigen, appropriately presented, and interleukin-2 (IL2), provided by T helper lymphocytes. We studied the specific CTL response induced by tumour cells transfected with murine IL2. Our results show that tumour cells manipulated to secrete IL2 induce an improved specific anti-tumour response which results in tumour rejection in mice. To further investigate the effect of IL2 on the CTL response to different antigens, we introduced synthetic peptides into IL2-secreting tumour cells and determined the specific CTL induction in syngeneic mice immunized with these cells. We report here that such IL2-secreting cells can effectively prime peptide-specific CTL in vivo....
Acta crystallographica. Section D, Biological crystallography, 2003
Three different crystal forms of the swine vesicular disease virus (SVDV), isolate SPA/2/'93,... more Three different crystal forms of the swine vesicular disease virus (SVDV), isolate SPA/2/'93, were obtained by the hanging-drop vapour-diffusion technique using ammonium sulfate and sodium/potassium phosphate as precipitants. Monoclinic crystals, space group C2, with unit-cell parameters a = 473.7, b = 385.3, c = 472.8 A, beta = 100.4 degrees, contain one virus pArticle in the crystal asymmetric unit and diffract to 3.0 A resolution. A second type of crystals had a cubic morphology and diffracted beyond 2.6 A resolution. These crystals belong to a primitive orthorhombic space group, with unit-cell parameters a = 319.6, b = 353.8, c = 377.7 A, and contain half a virus pArticle in the asymmetric unit. A third type of crystals, with a prismatic shape and belonging to space group I222, was also obtained under similar crystallization conditions. These latter crystals, with unit-cell parameters a = 318.3, b = 349.9, c = 371.7 A, diffract to at least 3.0 A resolution and contain 15 pro...
Developments in biological standardization, 1999
Foot-and-mouth disease virus (FMDV) has been one of the pioneering viral systems in the developme... more Foot-and-mouth disease virus (FMDV) has been one of the pioneering viral systems in the development of synthetic peptides as vaccines. Protection against FMDV infection is associated with the induction of neutralising antibodies. Therefore, attempts have been made to identify peptides capable of eliciting protective humoral responses. Peptides based on a continuous, immunodominant B cell site on the capsid protein VP1 have been shown to confer limited protection in natural hosts. This probably reflects the difficulties in reproducing the immunogenicity of an entire viral particle by using a much simpler synthetic antigen, due to: (i) the polymorphism of the class II MHC; (ii) the adequate presentation to the immune system of the peptides, and (iii) the difficulties of achieving protection against a highly variable RNA virus, which may favour selection of virus antigenic variants. The improvement of FMD peptide vaccines, and the development of in vitro alternatives to in vivo immunog...
The Journal of general virology, 1999
A recombinant live vector vaccine was produced by insertion of cDNA encoding the structural prote... more A recombinant live vector vaccine was produced by insertion of cDNA encoding the structural proteins (P1) of foot-and-mouth disease virus (FMDV) into a replication-competent human adenovirus type 5 vaccine strain (Ad5 wt). Groups of cattle (n = 3) were immunized twice, by the subcutaneous and/or intranasal routes, with either the Ad5 wt vaccine or with the recombinant FMDV Ad5-P1 vaccine. All animals were challenged by intranasal instillation of FMDV 4 weeks after the second immunizations. In the absence of a detectable antibody response to FMDV, significant protection against viral challenge was seen in all of the animals immunized twice by the subcutaneous route with the recombinant vaccine. The observed partial protection against clinical disease was not associated with a reduction in titre of persistent FMDV infections in the oropharynx of challenged cattle.
Tesis doctoral inédita de la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento d... more Tesis doctoral inédita de la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 20-10-1997 Bibliografía:h. 107-121
La enfermedad vesicular del cerdo es una enfermedad altamente contagiosa que afecta al ganado por... more La enfermedad vesicular del cerdo es una enfermedad altamente contagiosa que afecta al ganado porcino, causada por un enterovirus de la familia Picornaviridae. El virus es un derivado relativamente reciente del virus humano coxsackie-B5, con el que tiene gran homología molecular y antigénica. La enfermedad no es grave y los animales se suelen recuperar en pocos días. Sin embargo, suelen aparecer vesículas en el hocico y en los espacios interdigitales de las pezuñas. El hecho de que estos síntomas sean indistinguibles de los causados por el virus de la fiebre aftosa, hace que este virus esté incluido dentro de la lista A de la Oficina Internacional de Epizootias. Actualmente la enfermedad está erradicada en los países de la Unión Europea excepto en Italia, donde se considera endémica. Sin embargo, en Europa se mantiene el estado de alarma por la posible aparición de brotes, ya que esporádicamente aparecen en algunos países y han de ser erradicados inmediatamente. Para ello se ha invertido un considerable esfuerzo en investigación de métodos de diagnóstico eficaces y en el estudio del virus y la patología de la enfermedad, información que resumimos en esta revisión. PALABRAS CLAVE: Enfermedad vesicular EVC Virus Enterovirus Cerdo Diagnóstico Patología Respuesta inmune
Methods in Biotechnology, 2006
Page 1. 153 From: Methods in Biotechnology, Vol. 21: Food-Borne Pathogens: Methods and Protocols ... more Page 1. 153 From: Methods in Biotechnology, Vol. 21: Food-Borne Pathogens: Methods and Protocols Edited by: CC Adley © Humana Press Inc., Totowa, NJ 13 Detection of Enteroviruses Miguel-Angel Jiménez-Clavero, Victoria Ley, Nuria Gómez, and Juan-Carlos Sáiz ...
Virology, 2000
Swine vesicular disease virus (SVDV) is an enterovirus of the Picornaviridae family that belongs ... more Swine vesicular disease virus (SVDV) is an enterovirus of the Picornaviridae family that belongs to the coxsackievirus B group. A number of antigenic sites have been identified in SVDV by analysis of neutralizing monoclonal antibody-resistant mutants and shown to be exposed on the surface of the capsid. In this paper we have identified seven new immunodominant antigenic regions in SVDV capsid proteins by a peptide scanning method, using a panel of sera from infected pigs. When these antigenic regions were located in the capsid by using a computer-generated three-dimensional model of the virion, one was readily exposed on the surface of the virus and the remaining sites were located facing the inner side of the capsid shell, at subunit contacts, or in the interior of the subunit structure.
Virology, 1999
The importance of the induction of virus neutralizing antibodies to provide protection against fo... more The importance of the induction of virus neutralizing antibodies to provide protection against foot-and-mouth disease virus (FMDV) infection is well established. However, recent studies with recombinant adenovirus expressing the precursor polypeptide of the viral capsid (P1) indicate that cattle inoculated with this recombinant vector developed partial protection against FMDV infection, in the absence of a detectable specific humoral response. Other viral vectors have been widely used to induce protective immunity against many pathogens, and it has been reported that the use of different vectors for priming and boosting injections can provide a synergistic effect on this response. In this work, we determined the immunogenicity of two recombinant viruses (adenovirus and vaccinia) expressing P1-FMDV, administered either individually or sequentially, and the protection that they induced against FMDV challenge in pigs. A double immunization with the adeno-P1 virus was the most effective strategy at inducing protective immunity. In contrast to previous reports, the use of two different vectors for priming and boosting did not show a synergistic effect on the protection induced against FMD. Interestingly, immunized pigs developed FMDV-specific T cell responses but not detectable antibodies. Thus, the protection observed was likely to be mediated by a cellular immune response.
Virology, 1984
of African swine fever virus DNA (about 170 kbp) with the restriction endonucleases SolI, EcoRI, ... more of African swine fever virus DNA (about 170 kbp) with the restriction endonucleases SolI, EcoRI, Kpn1, PvuI, and SmaI yielded 14,31,17,13, and 11 fragments, respectively. The order of the restriction fragments produced by each nuclease was established by identifying the crosslinked EcoRI and Sal1 terminal fragments and then finding overlapping fragments. The five restriction fragment maps were integrated into a single map by locating SalI, KpnI, PvuI, and SvzaI sites in cloned EcoRI fragments, and orienting each fragment in the overall map.
Virology, 1984
African swine fever virus DNA (about 170 kbp) was cleaved with the restriction endonuclease EwRI ... more African swine fever virus DNA (about 170 kbp) was cleaved with the restriction endonuclease EwRI and most of the resulting 31 fragments were cloned in either the phage vector hWES.hB or the plasmid pBR325. Three fragments were not cloned in those vectors, the largest fragment EcoRI-A (21.2 kbp) and the two crosslinked terminal fragments, EcoRI-K' and D'. Endonuclease Sal1 cut fragment EcoRI-A into three pieces which were cloned in plasmid pBR322. The two terminal EcoRI fragments were cloned after removal of the crosslinks with nuclease Sl and addition of EcoRI linkers to the fragment ends. The complete library of the cloned fragments accounted for about 98% of ASF virus genome, the missing sequences being those removed by the nuclease Sl in the process of cloning the terminal fragments.
Molecular Immunology, 1986
Mouse monoclonal antibodies (MAbs) with different specificities against Dermatophagoides farinae ... more Mouse monoclonal antibodies (MAbs) with different specificities against Dermatophagoides farinae (D. farinae) extract have been obtained. Fifteen of these antibodies reacted with allergen molecules contained in D. farinae and D. pteronyssinus extracts, immunoprecipitating the main allergen of D. farinae (DF29) and homologous allergen of D. pteronyssinus (DP28). In addition, the monoclonal antibody MADF2 immunoprecipitated DF29 together with two other polypeptides (mol. wt 20,000 and 40,000) from D. farinae extracts. Five monoclonal antibodies (MADF2, MADF5, MADF9, MADF10 and MADF13) were selected to study their epitope specificity and the relationship of the epitope location on the allergen with the IgE binding site. By cross-inhibition studies two different epitopes and two partly overlapping determinants were found. In addition, two of these epitopes, those defined by MADF13 and MADF5, are close to, or overlapping, IgE binding site(s) on the allergen molecule. DF29 allergen from D. farinae extract was purified by affinity chromatography using MADF5 coupled to Sepharose. The purified allergen had capacity to bind mite specific human IgE and demonstrated an allergenic activity of up to 70% of total extract of D. farinae. These results indicate that DF29 molecule is the main allergen from D. farinae extracts.
Molecular Immunology, 1985
Allergen molecules from Purietaria judaica pollen, a widely distributed allergy inducer in Southe... more Allergen molecules from Purietaria judaica pollen, a widely distributed allergy inducer in Southern and Western Europe, have been studied using specific monoclonal antibodies (MAbs). MAbs