{"id":{"repo_id":"dialnet","oai_identifier":"oai:dialnet.unirioja.es:TES0000023209"},"canonical_url":"https://search.dev.ndltd.org/etd/dialnet/oai:dialnet.unirioja.es:TES0000023209","repository":{"repo_id":"dialnet","name":"Dialnet","base_url":"https://dialnet.unirioja.es/oaites/OAIHandler"},"display":{"title":"Development of an oral DNA anti-cancer vaccine targeting an angiogenic peptide, proadrenomedullin N-terminal 20 peptide (PAMP)","abstract":"Nucleic acid-based anticancer vaccines are becoming a very active field in the fight against cancer. The goal of this thesis was to generate an oral DNA vaccine targeting the angiogenic peptide, proadrenomedullin N-terminal 20 peptide (PAMP). Methods: An expression plasmid (PcPAMP) was generated by fusing the tetanus toxin epitopes P2 and P30 to the mouse PAMP sequence to counteract self-tolerance, and the empty plasmid was used as a negative control (PcNeg). The plasmids were introduced into Salmonella typhimurium bacteria that were then converted into bacterial ghosts. C57BL/6J mice were orally immunized with the ghosts five times at 2-week intervals. Then, B16-F10 melanoma cells were injected either into the tail vein to generate lung metastases or subcutaneously to generate cutaneous melanoma. Furthermore, naive CD4+ T cells were exposed to PAMP, and their secretome was analyzed by proximity extension assays. Results: In the lung metastasis model, a humoral and cellular immune response was evidenced by the presence of significantly higher amounts of anti-PAMP IgGs and spleen CD8+ T cells in the PcPAMP-vaccinated mice. Although the number and size of lung metastases were similar between both experimental groups, there was a notable reduction in intratumoral angiogenesis and cancer cell proliferation in the PcPAMP group. Furthermore, the treatment led to intense infiltration of lymphocytes, including regulatory T cells, and M2-like macrophages into the metastases. In the cutaneous melanoma model, similar results were obtained. Despite of a high level of anti-PAMP IgGs and a marked reduction of intratumoral angiogenesis in the PcPAMP-vaccinated mice, the treatment did not reduce tumor volume or improve survival rates. A remarkable infiltration of lymphocytes and macrophages was also observed in the tumors of vaccinated animals. Additionally, the incubation of naive CD4+ T cells with PAMP induced the up-regulation of IL-1β, IL-6, IL-7, IL-12, IL-27, TNFα, and FGF21, and the down-regulation of IL-16. Conclusions: Although the vaccine was not effective in reducing tumor growth, new proliferative and immune functions have been described for PAMP. These new functions include induction of melanoma proliferation and modulation of lymphocyte and macrophage tumor infiltration dynamics.","abstract_html":"Nucleic acid-based anticancer vaccines are becoming a very active field in the fight against cancer. The goal of this thesis was to generate an oral DNA vaccine targeting the angiogenic peptide, proadrenomedullin N-terminal 20 peptide (PAMP). Methods: An expression plasmid (PcPAMP) was generated by fusing the tetanus toxin epitopes P2 and P30 to the mouse PAMP sequence to counteract self-tolerance, and the empty plasmid was used as a negative control (PcNeg). The plasmids were introduced into Salmonella typhimurium bacteria that were then converted into bacterial ghosts. C57BL/6J mice were orally immunized with the ghosts five times at 2-week intervals. Then, B16-F10 melanoma cells were injected either into the tail vein to generate lung metastases or subcutaneously to generate cutaneous melanoma. Furthermore, naive CD4+ T cells were exposed to PAMP, and their secretome was analyzed by proximity extension assays. Results: In the lung metastasis model, a humoral and cellular immune response was evidenced by the presence of significantly higher amounts of anti-PAMP IgGs and spleen CD8+ T cells in the PcPAMP-vaccinated mice. Although the number and size of lung metastases were similar between both experimental groups, there was a notable reduction in intratumoral angiogenesis and cancer cell proliferation in the PcPAMP group. Furthermore, the treatment led to intense infiltration of lymphocytes, including regulatory T cells, and M2-like macrophages into the metastases. In the cutaneous melanoma model, similar results were obtained. Despite of a high level of anti-PAMP IgGs and a marked reduction of intratumoral angiogenesis in the PcPAMP-vaccinated mice, the treatment did not reduce tumor volume or improve survival rates. A remarkable infiltration of lymphocytes and macrophages was also observed in the tumors of vaccinated animals. Additionally, the incubation of naive CD4+ T cells with PAMP induced the up-regulation of IL-1β, IL-6, IL-7, IL-12, IL-27, TNFα, and FGF21, and the down-regulation of IL-16. Conclusions: Although the vaccine was not effective in reducing tumor growth, new proliferative and immune functions have been described for PAMP. These new functions include induction of melanoma proliferation and modulation of lymphocyte and macrophage tumor infiltration dynamics.","abstract_has_math":false,"creators":["Kalathil Raju, Tom"],"institution":"Universidad de La Rioja (España)","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Martínez Ramírez, Alfredo (Universidad de La Rioja)"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2025,"date_issued":"2025","date_published":"2025","updated_at":"2026-07-24T06:27:11Z","subjects":[],"languages":["eng"],"rights":["LICENCIA DE USO: Los documentos a texto completo incluidos en Dialnet son de acceso libre y propiedad de sus autores y/o editores. Por tanto, cualquier acto de reproducción, distribución, comunicación pública y/o transformación total o parcial requiere el consentimiento expreso y escrito de aquéllos. Cualquier enlace al texto completo de estos documentos deberá hacerse a través de la URL oficial de éstos en Dialnet. Más información: https://dialnet.unirioja.es/info/derechosOAI | INTELLECTUAL PROPERTY RIGHTS STATEMENT: Full text documents hosted by Dialnet are protected by copyright and/or related rights. This digital object is accessible without charge, but its use is subject to the licensing conditions set by its authors or editors. Unless expressly stated otherwise in the licensing conditions, you are free to linking, browsing, printing and making a copy for your own personal purposes. All other acts of reproduction and communication to the public are subject to the licensing conditions expressed by editors and authors and require consent from them. Any link to this document should be made using its official URL in Dialnet. More info: https://dialnet.unirioja.es/info/derechosOAI"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://dialnet.unirioja.es/servlet/oaites?codigo=396430","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Martínez Ramírez, Alfredo (Universidad de La Rioja)"]},{"key":"dc:creator","label":"Author","values":["Kalathil Raju, Tom"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2025"]},{"key":"dc:publisher","label":"Institution","values":["Universidad de La Rioja (España)"]},{"key":"dc:type","label":"Dc Type","values":["text (thesis)"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["LICENCIA DE USO: Los documentos a texto completo incluidos en Dialnet son de acceso libre y propiedad de sus autores y/o editores. Por tanto, cualquier acto de reproducción, distribución, comunicación pública y/o transformación total o parcial requiere el consentimiento expreso y escrito de aquéllos. Cualquier enlace al texto completo de estos documentos deberá hacerse a través de la URL oficial de éstos en Dialnet. Más información: https://dialnet.unirioja.es/info/derechosOAI | INTELLECTUAL PROPERTY RIGHTS STATEMENT: Full text documents hosted by Dialnet are protected by copyright and/or related rights. This digital object is accessible without charge, but its use is subject to the licensing conditions set by its authors or editors. Unless expressly stated otherwise in the licensing conditions, you are free to linking, browsing, printing and making a copy for your own personal purposes. All other acts of reproduction and communication to the public are subject to the licensing conditions expressed by editors and authors and require consent from them. Any link to this document should be made using its official URL in Dialnet. More info: https://dialnet.unirioja.es/info/derechosOAI"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://dialnet.unirioja.es/servlet/oaites?codigo=396430"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Nucleic acid-based anticancer vaccines are becoming a very active field in the fight against cancer. The goal of this thesis was to generate an oral DNA vaccine targeting the angiogenic peptide, proadrenomedullin N-terminal 20 peptide (PAMP). Methods: An expression plasmid (PcPAMP) was generated by fusing the tetanus toxin epitopes P2 and P30 to the mouse PAMP sequence to counteract self-tolerance, and the empty plasmid was used as a negative control (PcNeg). The plasmids were introduced into Salmonella typhimurium bacteria that were then converted into bacterial ghosts. C57BL/6J mice were orally immunized with the ghosts five times at 2-week intervals. Then, B16-F10 melanoma cells were injected either into the tail vein to generate lung metastases or subcutaneously to generate cutaneous melanoma. Furthermore, naive CD4+ T cells were exposed to PAMP, and their secretome was analyzed by proximity extension assays. Results: In the lung metastasis model, a humoral and cellular immune response was evidenced by the presence of significantly higher amounts of anti-PAMP IgGs and spleen CD8+ T cells in the PcPAMP-vaccinated mice. Although the number and size of lung metastases were similar between both experimental groups, there was a notable reduction in intratumoral angiogenesis and cancer cell proliferation in the PcPAMP group. Furthermore, the treatment led to intense infiltration of lymphocytes, including regulatory T cells, and M2-like macrophages into the metastases. In the cutaneous melanoma model, similar results were obtained. Despite of a high level of anti-PAMP IgGs and a marked reduction of intratumoral angiogenesis in the PcPAMP-vaccinated mice, the treatment did not reduce tumor volume or improve survival rates. A remarkable infiltration of lymphocytes and macrophages was also observed in the tumors of vaccinated animals. Additionally, the incubation of naive CD4+ T cells with PAMP induced the up-regulation of IL-1β, IL-6, IL-7, IL-12, IL-27, TNFα, and FGF21, and the down-regulation of IL-16. Conclusions: Although the vaccine was not effective in reducing tumor growth, new proliferative and immune functions have been described for PAMP. These new functions include induction of melanoma proliferation and modulation of lymphocyte and macrophage tumor infiltration dynamics.","Las vacunas anticancerígenas basadas en ácidos nucleicos se están convirtiendo en un campo muy activo en la lucha contra el cáncer. Nuestro objetivo en esta tesis ha sido generar una vacuna oral de DNA dirigida contra el péptido angiogénico de 20 aminoácidos derivado de la proadrenomedulina (PAMP). Métodos: Se generó un plásmido de expresión (PcPAMP) fusionando los epítopos P2 y P30 de la toxina tetánica a la secuencia del PAMP de ratón para contrarrestar la autotolerancia, y se utilizó el plásmido vacío como control negativo (PcNeg). Los plásmidos se introdujeron en bacterias Salmonella typhimurium que luego se transformaron en fantasmas bacterianos. Se utilizaron ratones C57BL/6J que fueron inmunizados por vía oral con los fantasmas en cinco ocasiones, con intervalos de dos semanas. A continuación, se inyectaron células de melanoma B16-F10 en la vena de la cola para generar metástasis pulmonares o se empleó la administración subcutánea para obtener un modelo de melanoma cutáneo. Además, se expusieron células T CD4+ a PAMP y se analizó su secretoma mediante ensayos de extensión de proximidad. Resultados: Se observó un aumento significativo en los niveles de IgG anti-PAMP y de células T CD8+ en el bazo de los ratones vacunados con PcPAMP, indicando un nivel suficiente de respuesta inmunitaria tanto humoral como celular. Aunque el número y el tamaño de las metástasis pulmonares fueron similares entre ambos grupos experimentales, se produjo una notable reducción de la angiogénesis intratumoral y de la proliferación de células cancerosas en el grupo PcPAMP. Además, el tratamiento provocó una intensa infiltración de linfocitos, incluidas células T reguladoras y macrófagos tipo M2, en las metástasis. En los casos de melanoma cutáneo se encontraron resultados similares. aA pesar de encontrarse niveles altos de IgGs anti-PAMP y una reducción de la angiogenesis intratumoral en los ratones vacunados con PcPAMP, el tratamiento tampoco redujo el volumen tumoral ni mejoró las tasas de supervivencia. En este modelo también se observó una fuerte infiltración de linfocitos y macrófagos en los ratones vacunados. Por otro lado, la incubación de células T CD4+ con PAMP indujo la regulación al alza de IL-1β, IL-6, IL-7, IL-12, IL-27, TNFα y FGF21, mientras que se observó una regulación a la baja para IL-16. Conclusiones: Aunque la vacuna no fue eficaz en la reducción del crecimiento tumoral, se han descrito nuevas funciones proliferativas e inmunes para PAMP. Estas nuevas funciones incluyen la inducción de la proliferación del melanoma y la modulación de la dinámica de infiltración tumoral de linfocitos y macrófagos"]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Development of an oral DNA anti-cancer vaccine targeting an angiogenic peptide, proadrenomedullin N-terminal 20 peptide (PAMP)"]}]}],"canonical_facts":{"dc:contributor":["Martínez Ramírez, Alfredo (Universidad de La Rioja)"],"dc:creator":["Kalathil Raju, Tom"],"dc:date":["2025"],"dc:description":["Nucleic acid-based anticancer vaccines are becoming a very active field in the fight against cancer. The goal of this thesis was to generate an oral DNA vaccine targeting the angiogenic peptide, proadrenomedullin N-terminal 20 peptide (PAMP). Methods: An expression plasmid (PcPAMP) was generated by fusing the tetanus toxin epitopes P2 and P30 to the mouse PAMP sequence to counteract self-tolerance, and the empty plasmid was used as a negative control (PcNeg). The plasmids were introduced into Salmonella typhimurium bacteria that were then converted into bacterial ghosts. C57BL/6J mice were orally immunized with the ghosts five times at 2-week intervals. Then, B16-F10 melanoma cells were injected either into the tail vein to generate lung metastases or subcutaneously to generate cutaneous melanoma. Furthermore, naive CD4+ T cells were exposed to PAMP, and their secretome was analyzed by proximity extension assays. Results: In the lung metastasis model, a humoral and cellular immune response was evidenced by the presence of significantly higher amounts of anti-PAMP IgGs and spleen CD8+ T cells in the PcPAMP-vaccinated mice. Although the number and size of lung metastases were similar between both experimental groups, there was a notable reduction in intratumoral angiogenesis and cancer cell proliferation in the PcPAMP group. Furthermore, the treatment led to intense infiltration of lymphocytes, including regulatory T cells, and M2-like macrophages into the metastases. In the cutaneous melanoma model, similar results were obtained. Despite of a high level of anti-PAMP IgGs and a marked reduction of intratumoral angiogenesis in the PcPAMP-vaccinated mice, the treatment did not reduce tumor volume or improve survival rates. A remarkable infiltration of lymphocytes and macrophages was also observed in the tumors of vaccinated animals. Additionally, the incubation of naive CD4+ T cells with PAMP induced the up-regulation of IL-1β, IL-6, IL-7, IL-12, IL-27, TNFα, and FGF21, and the down-regulation of IL-16. Conclusions: Although the vaccine was not effective in reducing tumor growth, new proliferative and immune functions have been described for PAMP. These new functions include induction of melanoma proliferation and modulation of lymphocyte and macrophage tumor infiltration dynamics.","Las vacunas anticancerígenas basadas en ácidos nucleicos se están convirtiendo en un campo muy activo en la lucha contra el cáncer. Nuestro objetivo en esta tesis ha sido generar una vacuna oral de DNA dirigida contra el péptido angiogénico de 20 aminoácidos derivado de la proadrenomedulina (PAMP). Métodos: Se generó un plásmido de expresión (PcPAMP) fusionando los epítopos P2 y P30 de la toxina tetánica a la secuencia del PAMP de ratón para contrarrestar la autotolerancia, y se utilizó el plásmido vacío como control negativo (PcNeg). Los plásmidos se introdujeron en bacterias Salmonella typhimurium que luego se transformaron en fantasmas bacterianos. Se utilizaron ratones C57BL/6J que fueron inmunizados por vía oral con los fantasmas en cinco ocasiones, con intervalos de dos semanas. A continuación, se inyectaron células de melanoma B16-F10 en la vena de la cola para generar metástasis pulmonares o se empleó la administración subcutánea para obtener un modelo de melanoma cutáneo. Además, se expusieron células T CD4+ a PAMP y se analizó su secretoma mediante ensayos de extensión de proximidad. Resultados: Se observó un aumento significativo en los niveles de IgG anti-PAMP y de células T CD8+ en el bazo de los ratones vacunados con PcPAMP, indicando un nivel suficiente de respuesta inmunitaria tanto humoral como celular. Aunque el número y el tamaño de las metástasis pulmonares fueron similares entre ambos grupos experimentales, se produjo una notable reducción de la angiogénesis intratumoral y de la proliferación de células cancerosas en el grupo PcPAMP. Además, el tratamiento provocó una intensa infiltración de linfocitos, incluidas células T reguladoras y macrófagos tipo M2, en las metástasis. En los casos de melanoma cutáneo se encontraron resultados similares. aA pesar de encontrarse niveles altos de IgGs anti-PAMP y una reducción de la angiogenesis intratumoral en los ratones vacunados con PcPAMP, el tratamiento tampoco redujo el volumen tumoral ni mejoró las tasas de supervivencia. En este modelo también se observó una fuerte infiltración de linfocitos y macrófagos en los ratones vacunados. Por otro lado, la incubación de células T CD4+ con PAMP indujo la regulación al alza de IL-1β, IL-6, IL-7, IL-12, IL-27, TNFα y FGF21, mientras que se observó una regulación a la baja para IL-16. Conclusiones: Aunque la vacuna no fue eficaz en la reducción del crecimiento tumoral, se han descrito nuevas funciones proliferativas e inmunes para PAMP. Estas nuevas funciones incluyen la inducción de la proliferación del melanoma y la modulación de la dinámica de infiltración tumoral de linfocitos y macrófagos"],"dc:format":["application/pdf"],"dc:identifier":["https://dialnet.unirioja.es/servlet/oaites?codigo=396430"],"dc:language":["eng"],"dc:publisher":["Universidad de La Rioja (España)"],"dc:rights":["LICENCIA DE USO: Los documentos a texto completo incluidos en Dialnet son de acceso libre y propiedad de sus autores y/o editores. Por tanto, cualquier acto de reproducción, distribución, comunicación pública y/o transformación total o parcial requiere el consentimiento expreso y escrito de aquéllos. Cualquier enlace al texto completo de estos documentos deberá hacerse a través de la URL oficial de éstos en Dialnet. Más información: https://dialnet.unirioja.es/info/derechosOAI | INTELLECTUAL PROPERTY RIGHTS STATEMENT: Full text documents hosted by Dialnet are protected by copyright and/or related rights. This digital object is accessible without charge, but its use is subject to the licensing conditions set by its authors or editors. Unless expressly stated otherwise in the licensing conditions, you are free to linking, browsing, printing and making a copy for your own personal purposes. All other acts of reproduction and communication to the public are subject to the licensing conditions expressed by editors and authors and require consent from them. Any link to this document should be made using its official URL in Dialnet. More info: https://dialnet.unirioja.es/info/derechosOAI"],"dc:title":["Development of an oral DNA anti-cancer vaccine targeting an angiogenic peptide, proadrenomedullin N-terminal 20 peptide (PAMP)"],"dc:type":["text (thesis)"]},"updated_at":"2026-07-24T06:27:11Z"}