Mejoramiento de una vacuna génica contra el Herpesvirus Bovino-1, uso de adyuvantes: Liposoma MAN?1-2MAN, CD40 ligando, MontanideTM GEL01 PR y Galectina-8
Bovine Herpesvirus type 1 (BoHV-1) has a worldwide distribution, causes various symptoms in cattle, is responsible for economic losses in the cattle industry and is recognized as an important component of the Bovine Respiratory Complex Disease (BRCD).\nAlthough inactivated and modified live virus (M...
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| Formato: | Tesis doctoral acceptedVersion |
| Lenguaje: | Español |
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Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica
2022
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| Acceso en línea: | http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=posgraafa&cl=CL1&d=HWA_6737 https://repositoriouba.sisbi.uba.ar/gsdl/collect/posgraafa/index/assoc/HWA_6737.dir/6737.PDF |
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I28-R145-HWA_6737 |
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dspace |
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Universidad de Buenos Aires |
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I-28 |
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R-145 |
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Repositorio Digital de la Universidad de Buenos Aires (UBA) |
| language |
Español |
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spa |
| topic |
Vacuna a ADN Adyuvante HVBo-1 Ratón Bovino Respuesta inmune Protección DNA vaccine Adjuvant BoHV-1 Mouse Bovine Immune response Protection Ciencias de la vida |
| spellingShingle |
Vacuna a ADN Adyuvante HVBo-1 Ratón Bovino Respuesta inmune Protección DNA vaccine Adjuvant BoHV-1 Mouse Bovine Immune response Protection Ciencias de la vida Kornuta, Claudia Alejandra Mejoramiento de una vacuna génica contra el Herpesvirus Bovino-1, uso de adyuvantes: Liposoma MAN?1-2MAN, CD40 ligando, MontanideTM GEL01 PR y Galectina-8 |
| topic_facet |
Vacuna a ADN Adyuvante HVBo-1 Ratón Bovino Respuesta inmune Protección DNA vaccine Adjuvant BoHV-1 Mouse Bovine Immune response Protection Ciencias de la vida |
| description |
Bovine Herpesvirus type 1 (BoHV-1) has a worldwide distribution, causes various symptoms in cattle, is responsible for economic losses in the cattle industry and is recognized as an important component of the Bovine Respiratory Complex Disease (BRCD).\nAlthough inactivated and modified live virus (MVL) vaccines are currently used, there are certain disadvantages in their use. In the case of inactivated vaccines, they do not provide complete protection and are generally poor in inducers of cellular immunity, while MVL have the risk of reversion to virulence, induction of abortions in pregnant cows and re-excretion of the vaccine strain by immunosuppression of cattle. The manipulation of the virus and the costs associated with its production are important points. On the other hand, DNA vaccines allow the differenciation infected to vaccinated animals (DIVA), are easy to produce, easy to transport and allow inducing humoral and cellular immune responses against specific antigens of the virus of interest.\nIn previous works, the use of a DNA-based vaccine encoding the truncated version of BoHV-1 glycoprotein D (pCIgD) together with chemical adjuvants to enhance DNA transfection has been shown to induce good levels of humoral and cellular immunity in the murine model, and partial protection in bovines.\nTo improve protection, the aim of this work is the evaluation of pCIgD in combination with different chemical and molecular adjuvants, first in mice and then in the natural host of the infection.\nThe adjuvants used were, first, a liposome decorated with MAN?1-2MAN molecules capable of specifically targeting its cargo to dendritic cell (DC) surface receptors called DC-SIGN. Then, a plasmid encoding the CD40L sequence that would allow triggering different responses by its interaction with CD40 present on DCs and other cells. Also, Galectin-8 (Gal-8) whose binding to cell surface glyco-receptors induces multiple cellular responses such as proliferation, differentiation, cytokine secretion among others, and the inclusion of the commercial adjuvant Montanide GEL01 PR. This latter, improves DNA transfection, recruits cells to the inoculation site and maintains the immune response for a longer period of time due to its "depot" effect.\nThe results showed that the use of the different adjuvants together with the DNA vaccine increased the immune response in both animal models. In humoral immunity, significant differences were obtained when total antibody titers, isotypes and neutralizing antibodies in serum and antibodies in mucosal were analyzed. Regarding cellular immunity, there was a significant increase in the proliferative response against BoHV-1 in animals vaccinated with the adjuvants compared to the response induced in animals vaccinated with pCIgD. In addition, upregulation of CD40, MHCII and CD86 molecules on the surface of bovine dendritic cells (DCs) was observed when cells were stimulated and activated with the adjuvants of the vaccine formulations.\nWhen virus challenge was performed, bovines vaccinated with the adjuvants elicited better protection, which was evidenced by lower viral excretion and clinical symptomatology. Undoubtedly, each adjuvant contributed to enhance the vaccine response due to its different chemical and/or molecular properties.\nThese adjuvants were formulated for the first time with a DNA vaccine and the results could be useful to design and improve a vaccine for the control of Infectious Bovine Rhinotracheitis (IBR). |
| author2 |
Langellotti, Cecilia Ana |
| author_facet |
Langellotti, Cecilia Ana Kornuta, Claudia Alejandra |
| format |
Tesis doctoral Tesis doctoral acceptedVersion |
| author |
Kornuta, Claudia Alejandra |
| author_sort |
Kornuta, Claudia Alejandra |
| title |
Mejoramiento de una vacuna génica contra el Herpesvirus Bovino-1, uso de adyuvantes: Liposoma MAN?1-2MAN, CD40 ligando, MontanideTM GEL01 PR y Galectina-8 |
| title_short |
Mejoramiento de una vacuna génica contra el Herpesvirus Bovino-1, uso de adyuvantes: Liposoma MAN?1-2MAN, CD40 ligando, MontanideTM GEL01 PR y Galectina-8 |
| title_full |
Mejoramiento de una vacuna génica contra el Herpesvirus Bovino-1, uso de adyuvantes: Liposoma MAN?1-2MAN, CD40 ligando, MontanideTM GEL01 PR y Galectina-8 |
| title_fullStr |
Mejoramiento de una vacuna génica contra el Herpesvirus Bovino-1, uso de adyuvantes: Liposoma MAN?1-2MAN, CD40 ligando, MontanideTM GEL01 PR y Galectina-8 |
| title_full_unstemmed |
Mejoramiento de una vacuna génica contra el Herpesvirus Bovino-1, uso de adyuvantes: Liposoma MAN?1-2MAN, CD40 ligando, MontanideTM GEL01 PR y Galectina-8 |
| title_sort |
mejoramiento de una vacuna génica contra el herpesvirus bovino-1, uso de adyuvantes: liposoma man?1-2man, cd40 ligando, montanidetm gel01 pr y galectina-8 |
| publisher |
Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica |
| publishDate |
2022 |
| url |
http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=posgraafa&cl=CL1&d=HWA_6737 https://repositoriouba.sisbi.uba.ar/gsdl/collect/posgraafa/index/assoc/HWA_6737.dir/6737.PDF |
| work_keys_str_mv |
AT kornutaclaudiaalejandra mejoramientodeunavacunagenicacontraelherpesvirusbovino1usodeadyuvantesliposomaman12mancd40ligandomontanidetmgel01prygalectina8 |
| _version_ |
1840330178371256320 |
| spelling |
I28-R145-HWA_67372025-06-25 Bovine Herpesvirus type 1 (BoHV-1) has a worldwide distribution, causes various symptoms in cattle, is responsible for economic losses in the cattle industry and is recognized as an important component of the Bovine Respiratory Complex Disease (BRCD).\nAlthough inactivated and modified live virus (MVL) vaccines are currently used, there are certain disadvantages in their use. In the case of inactivated vaccines, they do not provide complete protection and are generally poor in inducers of cellular immunity, while MVL have the risk of reversion to virulence, induction of abortions in pregnant cows and re-excretion of the vaccine strain by immunosuppression of cattle. The manipulation of the virus and the costs associated with its production are important points. On the other hand, DNA vaccines allow the differenciation infected to vaccinated animals (DIVA), are easy to produce, easy to transport and allow inducing humoral and cellular immune responses against specific antigens of the virus of interest.\nIn previous works, the use of a DNA-based vaccine encoding the truncated version of BoHV-1 glycoprotein D (pCIgD) together with chemical adjuvants to enhance DNA transfection has been shown to induce good levels of humoral and cellular immunity in the murine model, and partial protection in bovines.\nTo improve protection, the aim of this work is the evaluation of pCIgD in combination with different chemical and molecular adjuvants, first in mice and then in the natural host of the infection.\nThe adjuvants used were, first, a liposome decorated with MAN?1-2MAN molecules capable of specifically targeting its cargo to dendritic cell (DC) surface receptors called DC-SIGN. Then, a plasmid encoding the CD40L sequence that would allow triggering different responses by its interaction with CD40 present on DCs and other cells. Also, Galectin-8 (Gal-8) whose binding to cell surface glyco-receptors induces multiple cellular responses such as proliferation, differentiation, cytokine secretion among others, and the inclusion of the commercial adjuvant Montanide GEL01 PR. This latter, improves DNA transfection, recruits cells to the inoculation site and maintains the immune response for a longer period of time due to its "depot" effect.\nThe results showed that the use of the different adjuvants together with the DNA vaccine increased the immune response in both animal models. In humoral immunity, significant differences were obtained when total antibody titers, isotypes and neutralizing antibodies in serum and antibodies in mucosal were analyzed. Regarding cellular immunity, there was a significant increase in the proliferative response against BoHV-1 in animals vaccinated with the adjuvants compared to the response induced in animals vaccinated with pCIgD. In addition, upregulation of CD40, MHCII and CD86 molecules on the surface of bovine dendritic cells (DCs) was observed when cells were stimulated and activated with the adjuvants of the vaccine formulations.\nWhen virus challenge was performed, bovines vaccinated with the adjuvants elicited better protection, which was evidenced by lower viral excretion and clinical symptomatology. Undoubtedly, each adjuvant contributed to enhance the vaccine response due to its different chemical and/or molecular properties.\nThese adjuvants were formulated for the first time with a DNA vaccine and the results could be useful to design and improve a vaccine for the control of Infectious Bovine Rhinotracheitis (IBR). Fil: Kornuta, Claudia Alejandra. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Buenos Aires, Argentina Langellotti, Cecilia Ana Kornuta, Claudia Alejandra 2022-07-29 El Herpesvirus Bovino tipo 1 (HVBo-1) posee una amplia distribución mundial, es causante de diversos signos clínicos en el ganado, responsable de grandes pérdidas económicas en la industria ganadera y se lo reconoce como componente importante del Complejo Respiratorio Bovino (CRB).\nSi bien en la actualidad se utilizan vacunas a virus inactivado y a virus vivo modificado (MVL), existen ciertas desventajas en su uso. En el caso de las inactivadas, éstas no proporcionan una protección completa y son generalmente malas inductoras de la inmunidad celular, mientras que las MVL poseen riesgo de reversión a la virulencia, la inducción de abortos en vacas preñadas y reexcreción de la cepa vacunal por inmunosupresión del ganado. También hay que tener en cuenta la manipulación del virus y los costos asociados en su producción. En cambio, las vacunas a ADN permiten diferenciar animales vacunados de infectados (DIVA), son fáciles de producir, de transportar y permiten inducir respuestas inmunes humorales y celulares contra antígenos específicos de los virus de interés.\nEl uso de un plásmido que porta la secuencia de la gD del HVBo-1 (pCIgD) en su forma secretada junto con adyuvantes químicos, ha demostrado en trabajos previos inducir buenos niveles de inmunidad humoral y celular en el modelo murino, y protección parcial en bovinos.\nPara mejorar la protección, el objetivo de este trabajo es la evaluación del pCIgD en combinación con distintos adyuvantes químicos y moleculares, primero en ratones y luego en el huésped natural de la infección.\nLos adyuvantes empleados fueron un liposoma recubierto con moléculas de MAN?1-2MAN capaz de dirigir específicamente su carga a receptores de superficie de las células dendríticas (CD) denominados DC-SIGN, un plásmido que porta la secuencia del gen del CD40L que mejoraría la presentación por su interacción con el CD40 presente en las CD y otras células, Galectina-8 (Gal-8) cuya unión a glico-receptores de la superficie celular inducen múltiples respuestas celulares como proliferación, diferenciación, secreción de citoquinas, entre otras, y la inclusión del adyuvante comercial Montanide GEL01 PR que mejora la transfección del ADN y recluta células al sitio de inoculación y manteniendo una respuesta inmune por más tiempo debido a su efecto ?depot?.\nLos resultados mostraron que el uso de los distintos adyuvantes junto con la vacuna a ADN aumentaron la respuesta inmunitaria tanto del ratón como del bovino. En la inmunidad humoral, se obtuvieron diferencias significativas cuando se analizaron en suero los títulos de anticuerpos totales, los distintos isotipos de inmunoglobulinas y los anticuerpos con capacidad neutralizantes y los anticuerpos presentes en mucosa. En cuanto a la inmunidad celular, se produjo un aumento significativo de la respuesta proliferativa contra el HVBo-1 en los animales vacunados con los adyuvantes en comparación con la respuesta inducida en los animales vacunados con pCIgD solamente. Además, hubo regulación de las moléculas CD40, CMHII y CD86 en la superficie de las células dendríticas (CDs) bovinas cuando las células fueron estimuladas y activadas con los adyuvantes de las formulaciones de la vacuna.\nCuando se realizó el desafío viral, los bovinos vacunados con los adyuvantes obtuvieron una mejor protección, lo que se puso de manifiesto por una menor excreción del virus y signología clínica de los animales.\nEs la primera vez que estos adyuvantes fueron aplicados junto a una vacuna a ADN y los resultados podrían ser útiles para diseñar y mejorar una vacuna para el control de la rinotraqueitis infecciosa bovina. application/pdf Malchiodi, Emilio Mundo, Silvia Taboga, oscar Vacuna a ADN Adyuvante HVBo-1 Ratón Bovino Respuesta inmune Protección DNA vaccine Adjuvant BoHV-1 Mouse Bovine Immune response Protection spa Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/2.5/ar/ Ciencias de la vida Doctora de la Universidad de Buenos Aires en Ciencias Biológicas Mejoramiento de una vacuna génica contra el Herpesvirus Bovino-1, uso de adyuvantes: Liposoma MAN?1-2MAN, CD40 ligando, MontanideTM GEL01 PR y Galectina-8 info:eu-repo/semantics/doctoralThesis info:ar-repo/semantics/tesis doctoral info:eu-repo/semantics/acceptedVersion http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=posgraafa&cl=CL1&d=HWA_6737 https://repositoriouba.sisbi.uba.ar/gsdl/collect/posgraafa/index/assoc/HWA_6737.dir/6737.PDF |