A vaccine formulation combining rhoptry proteins NcROP40 and NcROP2 improves pup survival in a pregnant mouse model of neosporosis

Pastor-Fernández, Iván; Arranz-Solís, David; Regidor‐Cerrillo, Javier; Álvarez‐García, Gema; Hemphill, Andrew; García‐Culebras, Alicia; Cuevas-Martín, Carmen; Ortega‐Mora, Luis Miguel

doi:10.1016/j.vetpar.2014.12.009

Cited by 25 publications

(20 citation statements)

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“…However, worries about safety, resumed virulence, and increased numbers of carrier animals restricted their use in eld applications. In recent years, with the identi cation of new antigens, several dense granule (GRA) and rhoptry (ROP) proteins in N. caninum have been identi ed that could be used in diagnostics or as vaccine candidates [15,16,17]. Considering their safety and ease of preparation, vaccine development based on recombinant antigens offers many advantages over live-attenuated and inactivated vaccines.…”

Section: Discussionmentioning

confidence: 99%

Protective Immunity Against Neospora Caninum Infection Induced by 14-3-3 Protein in Mice

Zhang

Liu

et al. 2020

Preprint

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BackgroundNeospora caninum causes infections in a wide range of intermediate hosts and remains a threatening disease worldwide because of the lack of effective drugs and vaccines. Our previous studies demonstrated that N. caninum 14-3-3 protein (Nc14-3-3), which is included in N. caninum extracellular vesicles (NEVs), can induce effective immune responses and stimulate cytokine expression in mouse peritoneal macrophages. However, whether Nc14-3-3 has a protective effect and its mechanisms are poorly understood.MethodsHere, we evaluated immune responses and protective effects of Nc14-3-3 against 2×107 Nc-1 tachyzoites. Antibody (IgG, IgGl and IgG2a) levels and Th1-type (IFN-γ and IL-12) and Th2-type (IL-4 and IL-10) cytokines in mouse serum; survival rates; survival time; and parasite burdens were detected.ResultsIn the present study, the immunostimulatory effect of Nc14-3-3 was confirmed, as it triggered Th1-type cytokine (IFN-γ and IL-12) production in mouse serum two weeks after the final immunization. Moreover, the immunization of C57BL/6 mice with Nc14-3-3 induced high IgG antibody levels and significant increases in CD8+ T lymphocytes in the spleens of mice, indicating that a significant cellular immune response was induced. Mouse survival rates and survival times were significantly prolonged after immunization survival rates were 40% for Nc14-3-3 immunization and 60% for NEV immunization, while mice that received GST, PBS, or blank control all died at 13, 9, and 8 days after intraperitoneal N. caninum challenge. In addition, qPCR analysis indicated that there was a lower parasite burden and milder pathological changes in the mice immunized with Nc14-3-3.ConclusionsOur data demonstrate the vaccination of mice with Nc14-3-3 elicits both cellular and humoural immune responses and provides partial protection against acute neosporosis. Thus, Nc14-3-3 could be an effective antigen candidate for vaccine development for neosporosis.

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Section: Discussionmentioning

confidence: 99%

Protective Immunity Against Neospora Caninum Infection Induced by 14-3-3 Protein in Mice

Zhang

Liu

et al. 2020

Preprint

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“…rNcNTPase and rNcGRA7 proteins were obtained as previously described [ 30 ]. Briefly, proteins were cloned in the pET45b(+) expression system (Novagen), expressed in E. coli BL21(DE3) pLysS competent cells (Agilent Technologies) as a (His)6-tagged fusion proteins, and purified using HisTrapHP columns coupled to the ÄKTAprime Plus system (GE Healthcare).…”

Section: Methodsmentioning

confidence: 99%

The tandemly repeated NTPase (NTPDase) from Neospora caninum is a canonical dense granule protein whose RNA expression, protein secretion and phosphorylation coincides with the tachyzoite egress

et al. 2016

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BackgroundNTPases (also NTPDases) are enzymes with apyrase activity. They are widely distributed among eukaryotes, and also among members of the family Sarcocystidae. In Toxoplasma gondii, the TgNTPase accumulates in the dense granules, and has been commonly associated with the strain virulence. In the closely related Neospora caninum, the NcNTPase lacks nucleoside diphosphate hydrolase activity and appears to be more abundant in virulent isolates, indicating that it may contribute to the pathogenicity of neosporosis. However, so far no additional information on NcNTPase has been provided.MethodsHerein, the NcNTPase coding sequences were analysed by different in silico and de novo sequencing approaches. A comparative analysis of NcNTPase and NcGRA7 in terms of protein dynamics, secretion, phosphorylation, and mRNA expression profiles during the tachyzoite lytic cycle was also carried out. Moreover, NcNTPase immunolocalization was analysed by confocal microscopy techniques over a set number of time-points.ResultsWe describe the presence of three different loci containing three copies of the NcNTPase within the Nc-Liv genome, and report the existence of up to four different NcNTPase alleles in Nc-Liv. We also provide evidence for the occurrence of diverse protein species of the NcNTPase by two-dimensional gel electrophoresis. Both NcNTPase and NcGRA7 were similarly up-regulated and secreted during the egress and/or early invasion phases, and were phosphorylated. However, its secretion was not affected by the addition of calcium modulators such as A23187 and ethanol. NcNTPase and NcGRA7 localized in dense granules and parasitophorous vacuole membrane throughout the lytic cycle, although differed in their inmunolocalization during early invasion and egress.ConclusionsThe present study reveals the complexity of the NcNTPase loci in N. caninum. We hypothesize that the expression of different isoforms of the NcNTPase protein could contribute to parasite virulence. Our findings showed regulation of expression, secretion and phosphorylation of NcNTPase suggesting a potential role for progression through the tachyzoites lytic cycle.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1620-4) contains supplementary material, which is available to authorized users.

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“…Quil A was also used in an experimental vaccine against N. caninum , partially protecting mice against cerebral infection when s.c. co-administered with a N. caninum lysate (65). In a more recent research, Pastor-Fernández et al (55) showed that i.p. immunization with different recombinant antigens from N. caninum (rNcROP4, rNcROP2, rNcGRA7, and rNcNTPasa) as monovalent or pair-wise combinations (rNcROP40 + rNcROP2 and rNcGRA7 + rNcNTPase) + Quil A, increased specific immune responses, decreased parasite burden in brain and partially protected against vertical transmission in a pregnant mouse model of congenital neosporosis.…”

Section: Adjuvants: the Black Box Of Immunology Being Openedmentioning

confidence: 97%

Promising Plant-Derived Adjuvants in the Development of Coccidial Vaccines

2019

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Coccidial parasites cause medical and veterinary diseases worldwide, frequently leading to severe illness and important economic losses. At present, drugs, chemotherapeutics and prophylactic vaccines are still missing for most of the coccidial infections. Moreover, the development and administration of drugs and chemotherapeutics against these diseases would not be adequate in livestock, since they may generate unacceptable residues in milk and meat that would avoid their commercialization. In this scenario, prophylactic vaccines emerge as the most suitable approach. Subunit vaccines have proven to be biologically safe and economically viable, allowing researchers to choose among the best antigens against each pathogen. However, they are generally poorly immunogenic and require the addition of adjuvant compounds to the vaccine formulation. During the last decades, research involving plant immunomodulatory compounds has become an important field of study based on their potential pharmaceutical applications. Some plant molecules such as saponins, polysaccharides, lectins and heat shock proteins are being explored as candidates for adjuvant/carriers formulations. Moreover, plant-derived immune stimulatory compounds open the possibility to attain the main goal in adjuvant research: a safe and non-toxic adjuvant capable of strongly boosting and directing immune responses that could be incorporated into different vaccine formulations, including mucosal vaccines. Here, we review the immunomodulatory properties of several plant molecules and discuss their application and future perspective as adjuvants in the development of vaccines against coccidial infections.

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A vaccine formulation combining rhoptry proteins NcROP40 and NcROP2 improves pup survival in a pregnant mouse model of neosporosis

Cited by 25 publications

References 64 publications

Protective Immunity Against Neospora Caninum Infection Induced by 14-3-3 Protein in Mice

Protective Immunity Against Neospora Caninum Infection Induced by 14-3-3 Protein in Mice

The tandemly repeated NTPase (NTPDase) from Neospora caninum is a canonical dense granule protein whose RNA expression, protein secretion and phosphorylation coincides with the tachyzoite egress

Promising Plant-Derived Adjuvants in the Development of Coccidial Vaccines

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