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1.
J Transl Med ; 14(1): 224, 2016 07 28.
Artículo en Inglés | MEDLINE | ID: mdl-27465494

RESUMEN

BACKGROUND: High-risk human papillomaviruses (HR-HPVs) types 16 and 18 are the main etiological agents of cervical cancer, with more than 550,000 new cases each year worldwide. HPVs are also associated with other ano-genital and head-and-neck tumors. The HR-HPV E6 and E7 oncoproteins are responsible for onset and maintenance of the cell transformation state, and they represent appropriate targets for development of diagnostic and therapeutic tools. METHODS: The unmutated E6 gene from HPV16 and HPV18 and from low-risk HPV11 was cloned in a prokaryotic expression vector for expression of the Histidine-tagged E6 protein (His6-E6), according to a novel procedure. The structural properties were determined using circular dichroism and fluorescence spectroscopy. His6-E6 oncoprotein immunogenicity was assessed in a mouse model, and its functionality was determined using in vitro GST pull-down and protein degradation assays. RESULTS: The His6-tagged E6 proteins from HPV16, HPV18, and HPV11 E6 genes, without any further modification in the amino-acid sequence, were produced in bacteria as soluble and stable molecules. Structural analyses of HPV16 His6-E6 suggests that it maintains correct folding and conformational properties. C57BL/6 mice immunized with HPV16 His6-E6 developed significant humoral immune responses. The E6 proteins from HPV16, HPV18, and HPV11 were purified according to a new procedure, and investigated for protein-protein interactions. HR-HPV His6-E6 bound p53, the PDZ1 motif from MAGI-1 proteins, the human discs large tumor suppressor, and the human ubiquitin ligase E6-associated protein, thus suggesting that it is biologically active. The purified HR-HPV E6 proteins also targeted the MAGI-3 and p53 proteins for degradation. CONCLUSIONS: This new procedure generates a stable, unmutated HPV16 E6 protein, which maintains the E6 properties in in vitro binding assays. This will be useful for basic studies, and for development of diagnostic kits and immunotherapies in preclinical mouse models of HPV-related tumorigenesis.


Asunto(s)
Proteínas de Unión al ADN/biosíntesis , Mutación/genética , Neoplasias/diagnóstico , Neoplasias/terapia , Proteínas Oncogénicas Virales/biosíntesis , Infecciones por Papillomavirus/diagnóstico , Infecciones por Papillomavirus/terapia , Proteínas Recombinantes/biosíntesis , Proteínas Represoras/biosíntesis , Animales , Dicroismo Circular , Proteínas de Unión al ADN/aislamiento & purificación , Detergentes/farmacología , Femenino , Humanos , Inmunidad Humoral/efectos de los fármacos , Ratones Endogámicos C57BL , Chaperonas Moleculares/metabolismo , Neoplasias/virología , Proteínas Oncogénicas Virales/aislamiento & purificación , Unión Proteica/efectos de los fármacos , Desnaturalización Proteica/efectos de los fármacos , Proteolisis/efectos de los fármacos , Proteínas Represoras/aislamiento & purificación , Solubilidad
2.
J Transl Med ; 13: 80, 2015 Mar 05.
Artículo en Inglés | MEDLINE | ID: mdl-25763880

RESUMEN

BACKGROUND: Considering the high number of new cases of cervical cancer each year that are caused by human papilloma viruses (HPVs), the development of an effective vaccine for prevention and therapy of HPV-associated cancers, and in particular against the high-risk HPV-16 genotype, remains a priority. Vaccines expressing the E6 and E7 proteins that are detectable in all HPV-positive pre-cancerous and cancer cells might support the treatment of HPV-related lesions and clear already established tumors. METHODS: In this study, DNA and fowlpox virus recombinants expressing the E6F47R mutant of the HPV-16 E6 oncoprotein were generated, and their correct expression verified by RT-PCR, Western blotting and immunofluorescence. Immunization protocols were tested in a preventive or therapeutic pre-clinical mouse model of HPV-16 tumorigenicity using heterologous (DNA/FP) or homologous (DNA/DNA and FP/FP) prime/boost regimens. The immune responses and therapeutic efficacy were evaluated by ELISA, ELISPOT assays, and challenge with TC-1* cells. RESULTS: In the preventive protocol, while an anti-E6-specific humoral response was just detectable, a specific CD8(+) cytotoxic T-cell response was elicited in immunized mice. After the challenge, there was a delay in cancer appearance and a significant reduction of tumor volume in the two groups of E6-immunized mice, thus confirming the pivotal role of the CD8(+) T-cell response in the control of tumor growth in the absence of E6-specific antibodies. In the therapeutic protocol, in-vivo experiments resulted in a higher number of tumor-free mice after the homologous DNA/DNA or heterologous DNA/FP immunization. CONCLUSIONS: These data establish a preliminary indication for the prevention and treatment of HPV-related tumors by the use of DNA and avipox constructs as safe and effective immunogens following a prime/boost strategy. The combined use of recombinants expressing both E6 and E7 proteins might improve the antitumor efficacy, and should represent an important approach to control HPV-associated cancers.


Asunto(s)
Vacunas contra el Cáncer/inmunología , ADN Recombinante/metabolismo , Viruela Aviar/metabolismo , Papillomavirus Humano 16/inmunología , Inmunización Secundaria , Neoplasias/inmunología , Proteínas Oncogénicas Virales/genética , Proteínas Represoras/genética , Animales , Linfocitos T CD8-positivos/inmunología , Embrión de Pollo , Femenino , Humanos , Inmunidad Humoral/inmunología , Ratones Endogámicos C57BL , Neoplasias/patología , Transgenes , Vacunación , Replicación Viral
3.
Protein Pept Lett ; 31(7): 519-531, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-39021187

RESUMEN

BACKGROUND: Glycogen Storage Disease type III (GSD III) is a metabolic disorder resulting from a deficiency of the Glycogen Debranching Enzyme (GDE), a large monomeric protein (approximately 170 kDa) with cytoplasmic localization and two distinct enzymatic activities: 4-α-glucantransferase and amylo-α-1,6-glucosidase. Mutations in the Agl gene, with consequent deficiency in GDE, lead to the accumulation of abnormal/toxic glycogen with shorter chains (phosphorylase limit dextrin, PLD) in skeletal and/or heart muscle and/or in the liver. Currently, there is no targeted therapy, and available treatments are symptomatic, relying on specific diets. METHODS: Enzyme Replacement Therapy (ERT) might represent a potential therapeutic strategy for GSD III. Moreover, the single-gene nature of GSD III, the subcellular localization of GDE, and the type of affected tissues represent ideal conditions for exploring gene therapy approaches. Toward this direction, we designed a synthetic, codon-optimized cDNA encoding the human GDE. RESULTS: This gene yielded high amounts of soluble, enzymatically active protein in Escherichia coli. Moreover, when transfected in Human Embryonic Kidney cells (HEK-293), it successfully encoded a functional GDE. CONCLUSION: These results suggest that our gene or protein might complement the missing function in GSD III patients, opening the door to further exploration of therapeutic approaches for this disease.


Asunto(s)
Sistema de la Enzima Desramificadora del Glucógeno , Enfermedad del Almacenamiento de Glucógeno Tipo III , Humanos , Sistema de la Enzima Desramificadora del Glucógeno/genética , Sistema de la Enzima Desramificadora del Glucógeno/metabolismo , Enfermedad del Almacenamiento de Glucógeno Tipo III/genética , Enfermedad del Almacenamiento de Glucógeno Tipo III/terapia , Enfermedad del Almacenamiento de Glucógeno Tipo III/metabolismo , Enfermedad del Almacenamiento de Glucógeno Tipo III/enzimología , Células HEK293 , Terapia Genética/métodos , Terapia de Reemplazo Enzimático/métodos , Escherichia coli/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo
4.
J Transl Med ; 11: 95, 2013 Apr 11.
Artículo en Inglés | MEDLINE | ID: mdl-23578094

RESUMEN

BACKGROUND: The traditional smallpox vaccine, administered by scarification, was discontinued in the general population from 1980, because of the absence of new smallpox cases. However, the development of an effective prophylactic vaccine against smallpox is still necessary, to protect from the threat of deliberate release of the variola virus for bioterrorism and from new zoonotic infections, and to improve the safety of the traditional vaccine. Preventive vaccination still remains the most effective control and new vectors have been developed to generate recombinant vaccines against smallpox that induce the same immunogenicity as the traditional one. As protective antibodies are mainly directed against the surface proteins of the two infectious forms of vaccinia, the intracellular mature virions and the extracellular virions, combined proteins from these viral forms can be used to better elicit a complete and protective immunity. METHODS: Four novel viral recombinants were constructed based on the fowlpox genetic background, which independently express the vaccinia virus L1 and A27 proteins present on the mature virions, and the A33 and B5 proteins present on the extracellular virions. The correct expression of the transgenes was determined by RT-PCR, Western blotting, and immunofluorescence. RESULTS AND CONCLUSIONS: Using immunoprecipitation and Western blotting, the ability of the proteins expressed by the four novel FPL1R, FPA27L, FPA33R and FPB5R recombinants to be recognized by VV-specific hyperimmune mouse sera was demonstrated. By neutralisation assays, recombinant virus particles released by infected chick embryo fibroblasts were shown not be recognised by hyperimmune sera. This thus demonstrates that the L1R, A27L, A33R and B5R gene products are not inserted into the new viral progeny. Fowlpox virus replicates only in avian species, but it is permissive for entry and transgene expression in mammalian cells, while being immunologically non-cross-reactive with vaccinia virus. These recombinants might therefore represent safer and more promising immunogens that can circumvent neutralisation by vector-generated immunity in smallpox-vaccine-experienced humans.


Asunto(s)
Virus de la Viruela de las Aves de Corral/genética , Vacuna contra Viruela/genética , Vacunas Sintéticas/genética , Virus Vaccinia/genética , Vacunas Virales/genética , Animales , Embrión de Pollo , Chlorocebus aethiops , Fibroblastos/metabolismo , Genes Virales , Humanos , Ratones , Microscopía Fluorescente , Pruebas de Neutralización , Vacuna contra Viruela/inmunología , Transgenes , Vacunas Sintéticas/inmunología , Células Vero
5.
Protein Pept Lett ; 27(2): 145-157, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-31622193

RESUMEN

BACKGROUND: Glycogen storage disease type III (GSDIII, Cori/Forbes disease) is a metabolic disorder due to the deficiency of the Glycogen Debranching Enzyme (GDE), a large monomeric protein (about 176 kDa) with two distinct enzymatic activities: 4-α-glucantransferase and amylo-α-1,6-glucosidase. Several mutations along the amylo-alpha-1,6-glucosidase,4-alphaglucanotransferase (Agl) gene are associated with loss of enzymatic activity. The unique treatment for GSDIII, at the moment, is based on diet. The potential of plants to manufacture exogenous engineered compounds for pharmaceutical purposes, from small to complex protein molecules such as vaccines, antibodies and other therapeutic/prophylactic entities, was shown by modern biotechnology through "Plant Molecular Farming". OBJECTIVE AND METHODS: In an attempt to develop novel protein-based therapeutics for GSDIII, the Agl gene, encoding for the human GDE (hGDE) was engineered for expression as a histidinetagged GDE protein both in Nicotiana benthamiana plants by a transient expression approach, and in axenic hairy root in vitro cultures (HR) from Lycopersicum esculentum and Beta vulgaris. RESULTS: In both plant-based expression formats, the hGDE protein accumulated in the soluble fraction of extracts. The plant-derived protein was purified by affinity chromatography in native conditions showing glycogen debranching activity. CONCLUSION: These investigations will be useful for the design of a new generation of biopharmaceuticals based on recombinant GDE protein that might represent, in the future, a possible therapeutic option for GSDIII.


Asunto(s)
Sistema de la Enzima Desramificadora del Glucógeno/genética , Nicotiana/crecimiento & desarrollo , Raíces de Plantas/citología , Beta vulgaris/citología , Beta vulgaris/genética , Beta vulgaris/metabolismo , Técnicas de Cultivo de Célula , Cromatografía de Afinidad , Regulación de la Expresión Génica de las Plantas , Sistema de la Enzima Desramificadora del Glucógeno/aislamiento & purificación , Sistema de la Enzima Desramificadora del Glucógeno/metabolismo , Humanos , Solanum lycopersicum/citología , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Ingeniería de Proteínas , Proteínas Recombinantes/aislamiento & purificación , Nicotiana/genética , Nicotiana/metabolismo
6.
Hum Vaccin Immunother ; 13(2): 271-282, 2017 02.
Artículo en Inglés | MEDLINE | ID: mdl-28118086

RESUMEN

Signal sequences (ss) play a critical role in the sorting of nascent secretory and membrane proteins. This function has been conserved from bacteria through eukaryotes, although ss appear diverse in length and amino acid composition. Sorting of proteins is also critical to instruct antigens for a proper immunological response. Thus, a plant ss was used to drive Human Papillomavirus (HPV) model antigens into the human secretory pathway: the HPV16 E7 oncoprotein, its chimera with the coat protein (CP) of the Potato Virus X (PVX), the first 200 amino acids of the HPV16 minor capsid protein L2 (known to harbour cross-reacting epitopes) and its chimera with E7 gene. These genes were used to transfect HEK-293 cells and to immunize C57BL/6 mice. The ss-provided genes were expressed, and proteins detected by immunofluorescence and immunoblotting. Mouse immunization with DNA constructs carrying the ss elicited a strong humoral response against both E7 and L2 and a weak cell-mediated immunity. To our knowledge this is the first demonstration that a signal sequence derived from a plant can modulate the sorting of a heterologous protein in mammalian cells. This activity in mammalian cells may be responsible for the observed increased humoral response to DNA-based vaccines that are generally weak inducers of IgG response. This might open new perspectives in the design of DNA vaccines, especially to counteract infections where a strong humoral response is needed.


Asunto(s)
Inmunidad Humoral , Infecciones por Papillomavirus/prevención & control , Vacunas contra Papillomavirus/inmunología , Proteínas de Plantas/genética , Señales de Clasificación de Proteína , Vacunas de ADN/inmunología , Animales , Anticuerpos Antivirales/sangre , Proteínas de la Cápside/genética , Femenino , Inmunidad Celular , Ratones Endogámicos C57BL , Proteínas Oncogénicas Virales/genética , Proteínas E7 de Papillomavirus/genética , Vacunas contra Papillomavirus/genética , Potexvirus/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Resultado del Tratamiento , Vacunas de ADN/genética , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunología
7.
Antiviral Res ; 134: 182-191, 2016 10.
Artículo en Inglés | MEDLINE | ID: mdl-27637905

RESUMEN

The control of smallpox was achieved using live vaccinia virus (VV) vaccine, which successfully eradicated the disease worldwide. As the variola virus no longer exists as a natural infection agent, mass vaccination was discontinued after 1980. However, emergence of smallpox outbreaks caused by accidental or deliberate release of variola virus has stimulated new research for second-generation vaccine development based on attenuated VV strains. Considering the closely related animal poxviruses that also arise as zoonoses, and the increasing number of unvaccinated or immunocompromised people, a safer and more effective vaccine is still required. With this aim, new vectors based on avian poxviruses that cannot replicate in mammals should improve the safety of conventional vaccines, and protect from zoonotic orthopoxvirus diseases, such as cowpox and monkeypox. In this study, DNA and fowlpox (FP) recombinants that expressed the VV L1R, A27L, A33R, and B5R genes were generated (4DNAmix, 4FPmix, respectively) and tested in mice using novel administration routes. Mice were primed with 4DNAmix by electroporation, and boosted with 4FPmix applied intranasally. The lethal VVIHD-J strain was then administered by intranasal challenge. All of the mice receiving 4DNAmix followed by 4FPmix, and 20% of the mice immunized only with 4FPmix, were protected. The induction of specific humoral and cellular immune responses directly correlated with this protection. In particular, higher anti-A27 antibodies and IFNγ-producing T lymphocytes were measured in the blood and spleen of the protected mice, as compared to controls. VVIHD-J neutralizing antibodies in sera from the protected mice suggest that the prime/boost vaccination regimen with 4DNAmix plus 4FPmix may be an effective and safe mode to induce protection against smallpox and poxvirus zoonotic infections. The electroporation/intranasal administration routes contributed to effective immune responses and mouse survival.


Asunto(s)
Anticuerpos Neutralizantes/sangre , Electroporación , Viruela Aviar/genética , Vacuna contra Viruela/administración & dosificación , Vacunación/métodos , Virus Vaccinia/genética , Animales , Inmunidad Celular , Inmunidad Humoral , Interferón gamma/sangre , Interferón gamma/inmunología , Ratones , Mpox/prevención & control , Pruebas de Neutralización , Viruela/prevención & control , Vacuna contra Viruela/genética , Vacuna contra Viruela/inmunología , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunología , Virus Vaccinia/patogenicidad , Vacunas Virales/administración & dosificación , Vacunas Virales/genética , Vacunas Virales/inmunología
8.
Chemosphere ; 164: 627-633, 2016 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-27635645

RESUMEN

The emergence and diffusion of antibiotic-resistant bacteria has been a major public health problem for many years now. In this study, antibiotic-resistance of coliforms and Escherichia coli were investigated after their isolation from samples collected in a municipal wastewater treatment plant in the Milan area (Italy) along different points of the treatment sequence: inflow to biological treatment; outflow from biological treatment following rapid sand filtration; and outflow from peracetic acid disinfection. The presence of E. coli that showed resistance to ampicillin (AMP) and chloramphenicol (CAF), used as representative antibiotics for the efficacy against Gram-positive and Gram-negative bacteria, was evaluated. After determining E. coli survival using increasing AMP and CAF concentrations, specific single-resistant (AMPR or CAFR) and double-resistant (AMPR/CAFR) strains were identified among E. coli colonies, through amplification of the ß-lactamase Tem-1 (bla) and acetyl-transferase catA1 (cat) gene sequences. While a limited number of CAFR bacteria was observed, most AMPR colonies showed the specific resistance genes to both antibiotics, which was mainly due to the presence of the bla gene sequence. The peracetic acid, used as disinfection agent, showed to be very effective in reducing bacteria at the negligible levels of less than 10 CFU/100 mL, compatible with those admitted for the irrigation use of treated waters.


Asunto(s)
Antibacterianos/análisis , Farmacorresistencia Microbiana/efectos de los fármacos , Escherichia coli/aislamiento & purificación , Aguas Residuales , Contaminantes Químicos del Agua/análisis , Purificación del Agua/métodos , Antibacterianos/toxicidad , Farmacorresistencia Microbiana/genética , Escherichia coli/efectos de los fármacos , Escherichia coli/genética , Bacterias Gramnegativas/efectos de los fármacos , Italia , Pruebas de Sensibilidad Microbiana , Aguas Residuales/química , Aguas Residuales/microbiología , Contaminantes Químicos del Agua/toxicidad , beta-Lactamasas/genética
9.
Virus Res ; 225: 82-90, 2016 10 02.
Artículo en Inglés | MEDLINE | ID: mdl-27664839

RESUMEN

The therapeutic antitumor potency of a prime-boost vaccination strategy was explored, based on the mutated, nontransforming forms of the E6 (E6F47R) and E7 (E7GGG) oncogenes of Human Papilloma Virus type 16 (HPV16), fused to the Potato virus X (PVX) coat protein (CP) sequence. Previous data showed that CP fusion improves the immunogenicity of tumor-associated antigens and may thus increase their efficacy. After verifying the correct expression of E6F47RCP and E7GGGCP inserted into DNA and Fowlpox virus recombinants by Western blotting and immunofluorescence, their combined use was evaluated for therapy in a pre-clinical mouse model of HPV16-related tumorigenicity. Immunization protocols were applied using homologous (DNA/DNA) or heterologous (DNA/Fowlpox) prime-boost vaccine regimens. The humoral immune responses were determined by ELISA, and the therapeutic efficacy evaluated by the delay in tumor appearance and reduced tumor volume after inoculation of syngeneic TC-1* tumor cells. Homologous DNA/DNA genetic vaccines were able to better delay tumor appearance and inhibit tumor growth when DNAE6F47RCP and DNAE7GGGCP were administered in combination. However, the heterologous DNA/Fowlpox vaccination strategy was able to delay tumor appearance in a higher number of animals when E6F47RCP and in particular E7GGGCP were administered alone.


Asunto(s)
Proteínas de la Cápside/inmunología , Virus de la Viruela de las Aves de Corral/genética , Vectores Genéticos/genética , Proteínas Oncogénicas Virales/inmunología , Proteínas E7 de Papillomavirus/inmunología , Proteínas Recombinantes de Fusión/inmunología , Proteínas Represoras/inmunología , Vacunas de ADN/inmunología , Animales , Vacunas contra el Cáncer/inmunología , Proteínas de la Cápside/genética , Línea Celular , Modelos Animales de Enfermedad , Femenino , Expresión Génica , Humanos , Inmunización , Inmunización Secundaria , Ratones , Mutación , Proteínas Oncogénicas Virales/genética , Proteínas E7 de Papillomavirus/genética , Vacunas contra Papillomavirus/administración & dosificación , Vacunas contra Papillomavirus/genética , Vacunas contra Papillomavirus/inmunología , Transporte de Proteínas , Proteínas Recombinantes de Fusión/genética , Proteínas Represoras/genética , Vacunas de ADN/administración & dosificación , Vacunas de ADN/genética , Ensayos Antitumor por Modelo de Xenoinjerto
10.
Front Plant Sci ; 7: 54, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-26904039

RESUMEN

Severe acute respiratory syndrome (SARS) is a dangerous infection with pandemic potential. It emerged in 2002 and its aetiological agent, the SARS Coronavirus (SARS-CoV), crossed the species barrier to infect humans, showing high morbidity and mortality rates. No vaccines are currently licensed for SARS-CoV and important efforts have been performed during the first outbreak to develop diagnostic tools. Here we demonstrate the transient expression in Nicotiana benthamiana of two important antigenic determinants of the SARS-CoV, the nucleocapsid protein (N) and the membrane protein (M) using a virus-derived vector or agro-infiltration, respectively. For the M protein, this is the first description of production in plants, while for plant-derived N protein we demonstrate that it is recognized by sera of patients from the SARS outbreak in Hong Kong in 2003. The availability of recombinant N and M proteins from plants opens the way to further evaluation of their potential utility for the development of diagnostic and protection/therapy tools to be quickly manufactured, at low cost and with minimal risk, to face potential new highly infectious SARS-CoV outbreaks.

11.
Hum Vaccin Immunother ; 11(1): 45-52, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25483514

RESUMEN

Expression of HPV E5, E6 and E7 oncogenes are likely to overcome the regulation of cell proliferation and to escape immunological control, allowing uncontrolled growth and providing the potential for malignant transformation. Thus, their three oncogenic products may represent ideal target antigens for immunotherapeutic strategies. In previous attempts, we demonstrated that genetic vaccines against recombinant HPV16 E7 antigen were able to affect the tumor growth in a pre-clinical mouse model. To improve this anti-HPV strategy we developed a novel approach in which we explored the effects of E5-based genetic immunization. We designed novel HPV16 E5 genetic vaccines based on two different gene versions: whole E5 gene and E5Multi. The last one is a long multi epitope gene designed as a harmless E5 version. Both E5 genes were codon optimized for mammalian expression. In addition, we demonstrated that HPV 16 E5 oncogene is expressed in C3 mouse cell line making it an elective model for the study of E5 based vaccine. In this mouse model the immunological and biological activity of the E5 vaccines were assessed in parallel with the activity of anti-E7 and anti-E6 vaccines already reported to be effective in an immunotherapeutic setting. These E7 and E6 vaccines were made with mutated oncogenes, the E7GGG mutant that does not bind pRb and the E6F47R mutant that is less effective in inhibiting p53, respectively. Results confirmed the immunological activity of genetic formulations based on attenuated HPV16 oncogenes and showed that E5-based genetic immunization provided notable anti-tumor effects.


Asunto(s)
Papillomavirus Humano 16/inmunología , Proteínas Oncogénicas Virales/inmunología , Vacunas contra Papillomavirus/inmunología , Neoplasias del Cuello Uterino/prevención & control , Vacunas de ADN/inmunología , Animales , Línea Celular Tumoral , Femenino , Papillomavirus Humano 16/genética , Ratones Endogámicos C57BL , Proteínas Oncogénicas Virales/genética , Vacunas contra Papillomavirus/administración & dosificación , Vacunas contra Papillomavirus/genética , Vacunas de ADN/administración & dosificación , Vacunas de ADN/genética
12.
Virus Res ; 178(2): 374-82, 2013 Dec 26.
Artículo en Inglés | MEDLINE | ID: mdl-24050999

RESUMEN

The first-generation smallpox vaccine was based on live vaccinia virus (VV) and it successfully eradicated the disease worldwide. Therefore, it was not administered any more after 1980, as smallpox no longer existed as a natural infection. However, emerging threats by terrorist organisations has prompted new programmes for second-generation vaccine development based on attenuated VV strains, which have been shown to cause rare but serious adverse events in immunocompromised patients. Considering the closely related animal poxviruses that might also be used as bioweapons, and the increasing number of unvaccinated young people and AIDS-affected immunocompromised subjects, a safer and more effective smallpox vaccine is still required. New avipoxvirus-based vectors should improve the safety of conventional vaccines, and protect from newly emerging zoonotic orthopoxvirus diseases and from the threat of deliberate release of variola or monkeypox virus in a bioterrorist attack. In this study, DNA and fowlpox recombinants expressing the L1R, A27L, A33R and B5R genes were constructed and evaluated in a pre-clinical trial in mouse, following six prime/boost immunisation regimens, to compare their immunogenicity and protective efficacy against a challenge with the lethal VV IHD-J strain. Although higher numbers of VV-specific IFNγ-producing T lymphocytes were observed in the protected mice, the cytotoxic T-lymphocyte response and the presence of neutralising antibodies did not always correlate with protection. In spite of previous successful results in mice, rabbits and monkeys, where SIV/HIV transgenes were expressed by the fowlpox vector, the immune response elicited by these recombinants was low, and most of the mice were not protected.


Asunto(s)
Virus de la Viruela de las Aves de Corral/genética , Mpox/prevención & control , Vacuna contra Viruela/inmunología , Vacunas de ADN/inmunología , Virus Vaccinia/inmunología , Proteínas Virales/inmunología , Animales , Anticuerpos Neutralizantes/sangre , Anticuerpos Antivirales/sangre , Citotoxicidad Inmunológica , Femenino , Vectores Genéticos , Interferón gamma/metabolismo , Ratones , Ratones Endogámicos BALB C , Mpox/inmunología , Vacuna contra Viruela/administración & dosificación , Vacuna contra Viruela/genética , Linfocitos T/inmunología , Vacunas de ADN/administración & dosificación , Vacunas de ADN/genética , Virus Vaccinia/genética , Proteínas Virales/genética
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