المؤلفون: LANDRIEL, SOLEDAD CAMINATA, CASTILLO, JULIETA D.L.M., TABOGA, OSCAR A., FERRAROTTI, SUSANA A., GOTTLIEB, ALEXANDRA M., COSTA, HERNÁN

المصدر: Anais da Academia Brasileira de Ciências. January 2019 91(3)

مصطلحات موضوعية: Cyclodextrin glycosyltransferases, housekeeping genes, Paenibacillus, 16S rRNA

الوصف: Cyclodextrin glycosyltransferases (CGTases) are important enzymes in the biotechnology field because they catalyze starch conversion into cyclodextrins and linear oligosaccharides, which are used in food, pharmaceutical and cosmetic industries. The CGTases are classified according to their product specificity in α-, β-, α/β- and γ-CGTases. As molecular markers are the preferred tool for bacterial identification, we employed six molecular markers (16S rRNA, dnaK, gyrB, recA, rpoB and tufA) to test the identification of a CGTase-producing bacterial strain (DF 9R) in a phylogenetic context. In addition, we assessed the phylogenetic relationship of CGTases along bacterial evolution. The results obtained here allowed us to identify the strain DF 9R as Paenibacillus barengoltzii, and to unveil a complex origin for CGTase types during archaeal and bacterial evolution. We postulate that the α-CGTase activity represents the ancestral type, and that the γ-activity may have derived from β-CGTases.

وصف الملف: text/html

الوصول الحر: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652019000500620Test

المؤلفون: López, María Gabriela, López, Cinthia Ayelén, Gravisaco, María José, Alfonso, Victoria, Taboga, Oscar

المصدر: Archives of Virology; May2024, Vol. 169 Issue 5, p1-11, 11p

مستخلص: The occlusion bodies of Autographa californica multiple nucleopolyhedrovirus are proteinaceous formations with significant biotechnological potential owing to their capacity to integrate foreign proteins through fusion with polyhedrin, their primary component. However, the strategy for successful heterologous protein inclusion still requires further refinement. In this study, we conducted a comparative assessment of various conditions to achieve the embedding of recombinant proteins within polyhedra. Two baculoviruses were constructed: AcPHGFP (polh+), with GFP as a fusion to wild type (wt) polyhedrin and AcΔPHGFP (polh+), with GFP fused to a fragment corresponding to amino acids 19 to 110 of polyhedrin. These baculoviruses were evaluated by infecting Sf9 cells and stably transformed Sf9, Sf9POLH, and Sf9POLHE44G cells. The stably transformed cells contributed another copy of wt or a mutant polyhedrin, respectively. Polyhedra of each type were isolated and characterized by classical methods. The fusion PHGFP showed more-efficient incorporation into polyhedra than ΔPHGFP in the three cell lines assayed. However, ΔPHGFP polyhedron yields were higher than those of PHGFP in Sf9 and Sf9POLH cells. Based on an integral analysis of the studied parameters, it can be concluded that, except for the AcΔPHGFP/Sf9POLHE44G combination, deficiencies in one factor can be offset by improved performance by another. The combinations AcPHGFP/Sf9POLHE44G and AcΔPHGFP/Sf9POLH stand out due to their high level of incorporation and the large number of recombinant polyhedra produced, respectively. Consequently, the choice between these approaches becomes dependent on the intended application. [ABSTRACT FROM AUTHOR]

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المؤلفون: Masip, Yamil E., Caeiro, Lucas D., Cosenza, Maximiliano, Postan, Miriam, Molina, Guido, Taboga, Oscar, Molinari, María Paula, Tekiel, Valeria

المصدر: Frontiers in Cellular & Infection Microbiology; 2024, p01-16, 16p

مصطلحات موضوعية: TRYPANOSOMA cruzi, RECOMBINANT proteins, VACCINE effectiveness, VACCINATION, CHAGAS' disease, IMMUNOLOGIC memory

مستخلص: Chagas' is a neglected disease caused by the eukaryotic kinetoplastid parasite, Trypanosoma cruzi. Currently, approximately 8 million people are infected worldwide, most of whom are in the chronic phase of the disease, which involves cardiac, digestive, or neurologic manifestations. There is an urgent need for a vaccine because treatments are only effective in the initial phase of infection, which is generally underdiagnosed. The selection and combination of antigens, adjuvants, and delivery platforms for vaccine formulations should be designed to trigger mixed humoral and cellular immune responses, considering that T. cruzi has a complex life cycle with both intracellular and bloodstream circulating parasite stages in vertebrate hosts. Here, we report the effectiveness of vaccination with a T. cruzi-specfic protein family (TcTASV), employing both recombinant proteins with aluminum hydroxide and a recombinant baculovirus displaying a TcTASV antigen at the capsid. Vaccination stimulated immunological responses by producing lytic antibodies and antigen-specific CD4+ and CD8+ IFNγ secreting lymphocytes. More than 90% of vaccinated animals survived after lethal challenges with T. cruzi, whereas all control mice died before 30 days post-infection. Vaccination also induced a strong decrease in chronic tissue parasitism and generated immunological memory that allowed vaccinated and infected animals to control both the reactivation of the infection after immunosuppression and a second challenge with T. cruzi. Interestingly, inoculation with wild-type baculovirus partially protected the mice against T. cruzi. In brief, we demonstrated for the first time that the combination of the baculovirus platform and the TcTASV family provides effective protection against Trypanosoma cruzi, which is a promising vaccine for Chagas disease. [ABSTRACT FROM AUTHOR]

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المؤلفون: Amalfi, Sabrina, Plastine, María del Pilar, López, María Gabriela, Gravisaco, María José, Taboga, Oscar, Alfonso, Victoria

المصدر: Applied Microbiology & Biotechnology; Oct2023, Vol. 107 Issue 20, p6277-6286, 10p

مصطلحات موضوعية: TRANSGENE expression, GENES, CHO cell, CELL lines, GENETIC transduction, GENETIC vectors, VIRION

مستخلص: Poxins are poxviral proteins that act by degrading 2´3´-cGAMP, a key molecule of cGAS-STING axis that drives and amplifies the antiviral response. Previous works have described some poxin homologous among lepidopteran and baculoviral genes. In particular, P26, a poxin homologous from AcMNPV retains the 2´3´-cGAMP degradation activity in vitro. In this work, we demonstrated that the antiviral activity triggered by baculovirus was disrupted by the transient expression of P26 in murine and human cell lines, and the effect of this action is not only on IFN-β production but also on the induction of IFN-λ. Besides, we proved P26 functionality in a stable-transformed cell line where the protein was constitutively expressed, preventing the production of IFN-β induced by baculovirus and resulting in an improvement in the transduction efficiency by the attenuation of the antiviral activity. Finally, we incorporated P26 into budded virions by capsid display or passive incorporation, and the results showed that both strategies resulted in an improvement of 3–17 times in the efficiency of transgene expression in murine fibroblasts. Our results suggest that the incorporation of P26 to budded baculoviral vectors is a very promising tool to modulate negatively the innate antiviral cellular response and to improve the efficiency of gene delivery in mammalian cells. Key points: • P26 affects baculovirus-induced IFN-β and IFN–λ production in mammalian cells. • Murine fibroblasts expressing P26 are more susceptible to transduction by baculovirus. • Incorporation of P26 into the virion improves gene delivery efficiency of baculovirus. [ABSTRACT FROM AUTHOR]

: Copyright of Applied Microbiology & Biotechnology is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

المؤلفون: Leskow, Carla Coluccio, Kamenetzky, Laura, Dominguez, Pia Guadalupe, Zirpolo, José Antonio Díaz, Obata, Toshihiro, Costa, Hernán, Martí, Marcelo, Taboga, Oscar, Keurentjes, Joost, Sulpice, Ronan, Ishihara, Hirofumi, Stitt, Mark, Fernie, Alisdair Robert, Carrari, Fernando

المصدر: Journal of Experimental Botany, 2016 Jan 01. 67(14), 4091-4103.

الوصول الحر: https://www.jstor.org/stable/26391246Test

المؤلفون: Molinari, Paula, Crespo, María Inés, Molina, Guido N., Dho, Nicolás D., Marinho, Fábio V., Maletto, Belkys, Leclerc, Claude, Oliveira, Sergio C., Taboga, Oscar, Cebrian, Ignacio, Morón, Gabriel

المصدر: Immunology; May2023, Vol. 169 Issue 1, p27-41, 15p

مصطلحات موضوعية: CYTOTOXIC T cells, NUCLEOPOLYHEDROVIRUSES, ALFALFA looper, VIRAL proteins, MAJOR histocompatibility complex

مستخلص: Although the baculovirus Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) infects lepidopteran invertebrates as natural hosts, represents an efficient vector for vaccine development. Baculovirus surface display induces strong humoral responses against viruses and parasites. A novel strategy based on capsid display carrying foreign antigens in the AcMNPV particle further improved the immune response by eliciting CD8+ T cell activation. In this study, we analyze the intracellular mechanisms and signalling pathways involved in CD8+ T cell activation by capsid display. Our results show that baculovirus can attach to the cell surface, enter dendritic cells (DCs), transit within endocytic vesicles and escape to the cytosol for further degradation by the proteasome. We found that the availability of viral proteins, endosomal acidification, and proteasome activity are needed for efficient Major Histocompatibility Complex class‐I presentation by baculovirus carrying Ovalbumin in the viral capsid. Importantly, we demonstrated with this strategy that the induction of cytotoxic T cells and IL‐12 production by DCs are TLR9‐dependent and STING‐independent. Finally, our study shows differential intracellular processing for capsid and surface baculovirus proteins in DCs and highlights the role of different danger receptors during cytotoxic T cell priming through the capsid display delivery system, which could lead to improved baculovirus‐based vaccines development. [ABSTRACT FROM AUTHOR]

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المؤلفون: Smith, Ignacio, Mc Callum, Gregorio Juan, Sabljic, Adriana Victoria, Marfia, Juan Ignacio, Bombicino, Silvina Sonia, Trabucchi, Aldana, Iacono, Ruben Francisco, Birenbaum, Joaquín Manuel, Vazquez, Susana Claudia, Minoia, Juan Mauricio, Cascone, Osvaldo, López, María Gabriela, Taboga, Oscar, Targovnik, Alexandra Marisa, Wolman, Federico Javier, Fingermann, Matías, Alonso, Leonardo Gabriel, Valdez, Silvina Noemí, Miranda, María Victoria

المصدر: Biotechnology & Bioengineering; Oct2021, Vol. 118 Issue 10, p4129-4137, 9p

مستخلص: Serology testing for COVID‐19 is important in evaluating active immune response against SARS‐CoV‐2, studying the antibody kinetics, and monitoring reinfections with genetic variants and new virus strains, in particular, the duration of antibodies in virus‐exposed individuals and vaccine‐mediated immunity. In this study, recombinant S protein of SARS‐CoV‐2 was expressed in Rachiplusia nu, an important agronomic plague. One gram of insect larvae produces an amount of S protein sufficient for 150 determinations in the ELISA method herein developed. We established a rapid production process for SARS‐CoV‐2 S protein that showed immunoreactivity for anti‐SARS‐CoV‐2 antibodies and was used as a single antigen for developing the ELISA method with high sensitivity (96.2%) and specificity (98.8%). Our findings provide an efficient and cost‐effective platform for large‐scale S protein production, and the scale‐up is linear, thus avoiding the use of complex equipment like bioreactors. [ABSTRACT FROM AUTHOR]

: Copyright of Biotechnology & Bioengineering is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

المؤلفون: Currá, Anabella, Cacciabue, Marco, José Gravisaco, María, Asurmendi, Sebastián, Taboga, Oscar, Gismondi, María I.

المصدر: PeerJ; Jun2021, p1-18, 18p

مصطلحات موضوعية: FOOT & mouth disease, RNA replicase, VIRUS diseases, NON-coding RNA, MICRORNA, RNA polymerases

مستخلص: RNA interference (RNAi) is a well-conserved mechanism in eukaryotic cells that directs post-transcriptional gene silencing through small RNA molecules. RNAi has been proposed as an alternative approach for rapid and specific control of viruses including foot-and-mouth disease virus (FMDV), the causative agent of a devastating animal disease with high economic impact. The aim of this work was to assess the antiviral activity of different small RNA shuttles targeting the FMDV RNA-dependent RNA polymerase coding sequence (3D). Three target sequences were predicted within 3D considering RNA accessibility as a major criterion. The silencing efficacy of shorthairpin RNAs (shRNAs) and artificial microRNAs (amiRNAs) targeting the selected sequences was confirmed in fluorescent reporter assays. Furthermore, BHK-21 cells transiently expressing shRNAs or amiRNAs proved 70 to >95% inhibition of FMDV growth. Interestingly, dual expression of amiRNAs did not improve FMDV silencing. Lastly, stable cell lines constitutively expressing amiRNAs were established and characterized in terms of antiviral activity against FMDV. As expected, viral replication in these cell lines was delayed. These results show that the target RNA-accessibilityguided approach for RNAi design rendered efficient amiRNAs that constrain FMDV replication. The application of amiRNAs to complement FMDV vaccination in specific epidemiological scenarios shall be explored further. [ABSTRACT FROM AUTHOR]

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المؤلفون: Taboga Oscar, Molinari Paula, Peralta Andrea

المصدر: Virology Journal, Vol 6, Iss 1, p 192 (2009)

مصطلحات موضوعية: Infectious and parasitic diseases, RC109-216

الوصف: Abstract In order to improve the presentation and immunogenicity of single epitopes, virus-like particles (VLPs) are being used as platforms for the display of foreing epitopes on their surface. The rotavirus major capsid protein VP6 has the ability to self-assemble into empty non-infectious VLPs. In the present study, we analyzed the use of double layered VLPs (made up of VP2 and VP6 rotavirus proteins) as carriers to display a 14 amino acid epitope fused to three different aminoacidic regions of VP6 exposed on the surface of VLPs. Although all chimeric protein were correctly expressed in insect cells, only one of them resulted in spontaneous assembly of VLPs displaying the heterologous epitope on their surface, confirmed by sandwich ELISA and electron microscopy. Furthermore, the injection of chimeric VLPs into mice elicited higher antibody titers than the monomeric chimeric protein. Our results identify an specific amino acid region of VP6 which allows the insertion of at least a 14 amino acid heterolgous epitope and demonstrate its potential as immunogenic carrier.

وصف الملف: electronic resource

العلاقة: http://www.virologyj.com/content/6/1/192Test; https://doaj.org/toc/1743-422XTest

الوصول الحر: https://doaj.org/article/8e5784544ce14d809b1d784f8ae986e2Test

المؤلفون: Molinari, Paula, Molina, Guido N., Tavarone, Eugenia, Del Médico Zajac, María Paula, Morón, Gabriel, Taboga, Oscar

المصدر: Applied Microbiology & Biotechnology; Dec2018, Vol. 102 Issue 23, p10139-10146, 8p

مصطلحات موضوعية: BACULOVIRUSES, BACULOVIRUS diseases, VACCINATION, IMMUNIZATION, ANTIGEN synthesis

مستخلص: The baculovirus Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) infects lepidopteran invertebrates as natural hosts, although it also has been used as display vector for vaccine development. In this work, we evaluated the effectiveness of repetitive doses of AcMNPV-based vectors on the cytotoxic immune response specific to the capsid-displayed heterologous antigen ovalbumin (OVA). Our results demonstrate that baculovirus vectors induce a boosting effect in the cytotoxic immune response to OVA, making possible to recover the levels obtained in the primary response. Moreover, mice preimmunized with wild-type baculovirus showed a complete lack of antigen-specific CD8 cytotoxic T lymphocytes (CTLs) that may be related to the presence of antibodies directed to baculoviral surface proteins, particularly to GP64. However, baculovirus was able to induce the innate immune response in spite of a previous response against this vector, although some quantitative differences reflect a distinct activation of the immune cells in prime and boost. This is the first report in which the novel capsid display strategy is evaluated in prime-boost schemes to improve efficient CTL responses. [ABSTRACT FROM AUTHOR]

: Copyright of Applied Microbiology & Biotechnology is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)