Oral Immunization With a Plant HSP90-SAG1 Fusion Protein Produced in Tobacco Elicits Strong Immune Responses and Reduces Cyst Number and Clinical Signs of Toxoplasmosis in Mice

Sánchez-López, Edwin; Corigliano, Mariana G.; Oliferuk, Sonia; Rivera, Maximiliano; Mendoza-Morales, Luisa F; Angel, Sérgio O.; Sander, Valeria Analía; Clemente, Marina

doi:10.3389/fpls.2021.726910

Cited by 12 publications

(9 citation statements)

References 125 publications

(192 reference statements)

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“…It is relevant that these tachyzoites were able to generate a complete immune response, which only happens when cells are invaded by tachyzoites and not with an infection with dead or lysed tachyzoites [85][86][87][88]. In this sense, Toxovac® is a live attenuated vaccine used to prevent sheep congenital toxoplasmosis [89].…”

Section: Discussionmentioning

confidence: 99%

“…Western blot was then performed as described [90]. The primary antibodies: αrH2B.Z [22], αrH2A.Z [23], αrROP5 (rabbit) and αrROP18 (rabbit) were used at 1/5000, whereas α-c-Myc (rabbit, abCam ab9106) was used at 1/2000 and αrSag1 [87] 1/200 for 1 h at room temperature. αH3ac (rabbit, Millipore, 06-599B) 1/200, was incubated overnight.…”

Section: Methodsmentioning

confidence: 99%

“…Samples were boiled for 5 min and 20 µl were loaded per well in a 15% SDS-PAGE gel for immunoblotting. The absence of contaminating proteins was corroborated by Western blot with murine anti-SAG1antibody [87]. Quantification of the bands was done with Image J 1.53q (Fiji) software.…”

Section: Methodsmentioning

confidence: 99%

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Histone variant H2B.Z acetylation is necessary for maintenance ofToxoplasma gondiibiological fitness

Vanagas

Muñoz

Cristaldi

et al. 2023

Preprint

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Through regulation of DNA packaging, histone proteins are fundamental to a wide array of biological processes. A variety of post-translational modifications (PTMs), including acetylation, constitute a proposed histone code that is interpreted by reader proteins to modulate chromatin structure. Canonical histones can be replaced with variant versions that add an additional layer of regulatory complexity. The protozoan parasite Toxoplasma gondii is unique among eukaryotes in possessing a novel variant of H2B designated H2B.Z. The combination of PTMs and the use of histone variants is important for gene regulation in T. gondii, offering new targets for drug development. In this work, T. gondii parasites were generated in which the 5 N-terminal acetylatable lysines in H2B.Z were mutated to either alanine (c-Myc-A) or arginine (c-Myc-R). c-Myc-A mutant only displayed a mild effect in its ability to kill mice. c-Myc-R mutant presented an impaired ability to grow and an increase in differentiation to latent bradyzoites. This mutant line was also more sensitive to DNA damage, displayed no virulence in mice, and provided protective immunity against future infection. While nucleosome composition was unaltered, key genes were abnormally expressed during in vitro bradyzoite differentiation. Our results show that the N-terminal positive charge patch of H2B.Z is important for these procceses. Pull down assays with acetylated N-terminal H2B.Z peptide and unacetylated one retrieved common and differential interactors. Acetylated peptide pulled down proteins associated with chromosome maintenance/segregation and cell cycle, opening the question of a possible link between H2B.Z acetylation status and mitosis.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Histone variant H2B.Z acetylation is necessary for maintenance ofToxoplasma gondiibiological fitness

Vanagas

Muñoz

Cristaldi

et al. 2023

Preprint

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“…However, it is more laborious, and the expression levels of the recombinant proteins vary from case to case. In this case, it was also shown to be highly efficient in generating an adequate immune response[7,13,17].…”

Section: Discussionmentioning

confidence: 99%

Safe plant Hsp90 adjuvants elicit an effective immune response against SARS-CoV2 derived RBD antigen

Ramos-Duarte,

Orlowski,

Jaquenod de Giusti

et al. 2023

Preprint

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To better understand the role of pHsp90 adjuvant in immune response modulation, we proposed the use of the Receptor Binding Domain (RBD) of the Spike protein of SARS-CoV2, the principal candidate in the design of subunit vaccines. We evaluated the humoral and cellular immune responses against RBD through the strategy “protein mixture” (Adjuvant + Antigen). The rRBD adjuvanted with rAtHsp81.2 group showed a higher increase of the anti-rRBD IgG1, while the rRBD adjuvanted with rNbHsp90.3 group showed a significant increase of anti-rRBD IgG2b/2a. These results were consistent with the cellular immune response analysis. Spleen cell cultures from rRBD+rNbHsp90.3-immunized mice showed significantly increased IFN-γ production. In contrast, spleen cell cultures from rRBD+rAtHsp81.2-immunized mice showed significant increased IL-4 levels. Finally, vaccines adjuvanted with rNbHsp90.3 induced higher neutralizing antibody responses compared to those adjuvanted with rAtHsp81.2. To know whether both chaperones must form complexes to generate an effective immune response, we performed co-immunoprecipitation (co-IP) assays. The results indicated that the greater neutralizing capacity observed in the rRBD adjuvanted with rNbHsp90.3 group would be given by the rRBD-rNbHsp90.3 interaction rather than by the quality of the immune response triggered by the adjuvants. These results, together with our previous results, provide a comparative benchmark of these two novel and safe vaccine adjuvants for their capacity to stimulate immunity to a subunit vaccine, demonstrating the capacity of adjuvanted SARS-CoV2 subunit vaccines. Furthermore, these results revealed differences in the ability to modulate the immune response between these two pHsp90s, highlighting the importance of adjuvant selection for future rational vaccine and adjuvant design.

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“…It has been used as a potential target for immune response and shown to provide protection against Eimeria infection as DNA or subunit vaccines (Pagheh et al, 2020). Enhancements to its immune protective capacity have been achieved through fusion or co-administration with certain proteins including IL-2, IL-18, HSP90, et al (Liu et al, 2010;Sanchez-Lopez et al, 2021;Song et al, 2009;Xu et al, 2008). In addition, multiple antigen delivery systems, including PLGA nanoparticle, Salmonella typhimurium, Lactobacillus plantarum, improve its protective effect (Allahyari et al, 2022;Liu et al, 2020;Pogonka et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Recombinant production of SAG1 fused with xylanase in Pichia pastoris induced higher protective immunity against Eimeria tenella infection in chicken

Liu,

Wei,

Liang

et al. 2024

Microbial Biotechnology

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Chicken coccidiosis is an intestinal disease caused by the parasite Eimeria, which severely damages the growth of chickens and causes significant economic losses in the poultry industry. Improvement of the immune protective effect of antigens to develop high efficiency subunit vaccines is one of the hotspots in coccidiosis research. Sporozoite‐specific surface antigen 1 (SAG1) of Eimeria tenella (E. tenella) is a well‐known protective antigen and is one of the main target antigens for the development of subunit, DNA and vector vaccines. However, the production and immunoprotective effects of SAG1 need to be further improved. Here, we report that both SAG1 from E. tenella and its fusion protein with the xylanase XynCDBFV‐SAG1 are recombinant expressed and produced in Pichia pastoris (P. pastoris). The substantial expression quantity of fusion protein XynCDBFV‐SAG1 is achieved through fermentation in a 15‐L bioreactor, reaching up to about 2 g/L. Moreover, chickens immunized with the fusion protein induced higher protective immunity as evidenced by a significant reduction in the shedding of oocysts after E. tenella challenge infection compared with immunized with recombinant SAG1. Our results indicate that the xylanase enhances the immunogenicity of subunit antigens and has the potential for developing novel molecular adjuvants. The high expression level of fusion protein XynCDBFV‐SAG1 in P. pastoris holds promise for the development of effective recombinant anti‐coccidial subunit vaccine.

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Oral Immunization With a Plant HSP90-SAG1 Fusion Protein Produced in Tobacco Elicits Strong Immune Responses and Reduces Cyst Number and Clinical Signs of Toxoplasmosis in Mice

Cited by 12 publications

References 125 publications

Histone variant H2B.Z acetylation is necessary for maintenance ofToxoplasma gondiibiological fitness

Histone variant H2B.Z acetylation is necessary for maintenance ofToxoplasma gondiibiological fitness

Safe plant Hsp90 adjuvants elicit an effective immune response against SARS-CoV2 derived RBD antigen

Recombinant production of SAG1 fused with xylanase in Pichia pastoris induced higher protective immunity against Eimeria tenella infection in chicken

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