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A Toxoplasma gondii vaccine encoding multistage antigens in conjunction with ubiquitin confers protective immunity to BALB/c mice against parasite infection.

Yin H, Zhao L, Wang T, Zhou H, He S, Cong H - Parasit Vectors (2015)

Bottom Line: DNA vaccines have proved effective in the protection against parasites.Our results indicated that the DNA vaccine had the advantage of inducing a stronger humoral response, whereas the adenovirus-vectored vaccine effectively improved the cellular immune response.Priming vaccination with DNA vaccine and boosting with the recombinant adenovirus vaccine encoding ubiquitin conjugated multi-stage antigens of T. gondii was proved to be a potential strategy against the infection of type I and type II parasite.

View Article: PubMed Central - PubMed

Affiliation: Department of Human Parasitology, Shandong University, School of Medicine, No. 44 Wenhuaxi Road, Jinan, Shandong, 250012, PR China. yinhuiquan521@126.com.

ABSTRACT

Background: Toxoplasma gondii is a widely prevalent intracellular parasite which infects almost all warm-blooded animals including humans and causes serious zoonotic toxoplasmosis. DNA vaccines have proved effective in the protection against parasites. However, the problems of weak immunity and inefficient delivery of DNA vaccine remain major issues. Therefore, comprehensive antigens derived from all stages of the parasite, effective adjuvants and delivery systems should be considered in the vaccine construction.

Methods: SAG3101-144,ROP18347-396, MIC6288-347, GRA7182-224, MAG158-125, BAG1156-211 and SPA142-200, derived from antigens in tachyzoite, bradyzoite and sporozoite stages of T. gondii were screened based on CD8(+) T cell epitope binding affinity to HLA and H-2. We constructed a recombinant DNA vaccine and an adenovirus vaccine encoding multi-stage antigen of T. gondii linked to ubiquitin molecules and vaccinated BALB/c mice with different strategies. Antibodies, cytokines, splenocytes proliferation, as well as the percentage of CD4(+) and CD8(+) T cells in immunized mouse were analyzed by the Enzyme-Linked Immunosorbent Assays (ELISA), Flow Cytometry (FCM). Protective efficacy was evaluated by challenging immunized mice with type I and type II parasite.

Results: Our results indicated that the DNA vaccine had the advantage of inducing a stronger humoral response, whereas the adenovirus-vectored vaccine effectively improved the cellular immune response. Priming with DNA vaccine and boosting with adenovirus-vectored vaccine induced Th1-type immune responses with highest levels of IgG2a and secretion of cytokines IL-2 and IFN-γ. Effective protection against type I and type II parasite with an increase in survival rate and a decrease in brain cyst burden was achieved in immunized mice.

Conclusions: Priming vaccination with DNA vaccine and boosting with the recombinant adenovirus vaccine encoding ubiquitin conjugated multi-stage antigens of T. gondii was proved to be a potential strategy against the infection of type I and type II parasite.

No MeSH data available.


Related in: MedlinePlus

Construction of DNA and adenovirus vaccines encoding ubiquitin-conjugated multistage antigen segments derived from T. gondii. a Schematic representation of the compound proteins, which encode amino acid sequences of eight protein segments derived from tachyzoite, bradyzoite and sporozoite antigens of T. gondii. b Construction of recombinant DNA and adenovirus vaccines expressing ubiquitin-conjugated multistage antigen segments. The recombinant eukaryotic plasmids pVAX1-MAS (c), pVAX1-UMAS (d), and pHBAd-MCMV-GFP-UMSA (e) were identified by restriction enzyme digestion analysis (Lane 1). Lane M: DNA marker. f Recombinant adenoviral particles were obtained from 293 cells
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Fig1: Construction of DNA and adenovirus vaccines encoding ubiquitin-conjugated multistage antigen segments derived from T. gondii. a Schematic representation of the compound proteins, which encode amino acid sequences of eight protein segments derived from tachyzoite, bradyzoite and sporozoite antigens of T. gondii. b Construction of recombinant DNA and adenovirus vaccines expressing ubiquitin-conjugated multistage antigen segments. The recombinant eukaryotic plasmids pVAX1-MAS (c), pVAX1-UMAS (d), and pHBAd-MCMV-GFP-UMSA (e) were identified by restriction enzyme digestion analysis (Lane 1). Lane M: DNA marker. f Recombinant adenoviral particles were obtained from 293 cells

Mentions: Recombinant adenovirus expressing UMAS (Ad-UMAS) was generated using the AdMax^TM system (Hanbio, Shanghai, China) by homologous recombination of pHBAd-MCMV-GFP-UMAS with pHBAd-BHG in HEK-293 cells. The titre of 1011 plaque-forming units (PFU)/mL Ad-UMAS particles was purified by cesium chloride gradient centrifugation and then stored in storage buffer (10 mM Tris, 2 mM MgCl2, 5 % sucrose, pH 8.0) at–80°C. Figure 1 shows the construction of the DNA vaccines (p-MAS and p-UMAS) and adenovirus vaccine (Ad-UMAS).Fig. 1


A Toxoplasma gondii vaccine encoding multistage antigens in conjunction with ubiquitin confers protective immunity to BALB/c mice against parasite infection.

Yin H, Zhao L, Wang T, Zhou H, He S, Cong H - Parasit Vectors (2015)

Construction of DNA and adenovirus vaccines encoding ubiquitin-conjugated multistage antigen segments derived from T. gondii. a Schematic representation of the compound proteins, which encode amino acid sequences of eight protein segments derived from tachyzoite, bradyzoite and sporozoite antigens of T. gondii. b Construction of recombinant DNA and adenovirus vaccines expressing ubiquitin-conjugated multistage antigen segments. The recombinant eukaryotic plasmids pVAX1-MAS (c), pVAX1-UMAS (d), and pHBAd-MCMV-GFP-UMSA (e) were identified by restriction enzyme digestion analysis (Lane 1). Lane M: DNA marker. f Recombinant adenoviral particles were obtained from 293 cells
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4588682&req=5

Fig1: Construction of DNA and adenovirus vaccines encoding ubiquitin-conjugated multistage antigen segments derived from T. gondii. a Schematic representation of the compound proteins, which encode amino acid sequences of eight protein segments derived from tachyzoite, bradyzoite and sporozoite antigens of T. gondii. b Construction of recombinant DNA and adenovirus vaccines expressing ubiquitin-conjugated multistage antigen segments. The recombinant eukaryotic plasmids pVAX1-MAS (c), pVAX1-UMAS (d), and pHBAd-MCMV-GFP-UMSA (e) were identified by restriction enzyme digestion analysis (Lane 1). Lane M: DNA marker. f Recombinant adenoviral particles were obtained from 293 cells
Mentions: Recombinant adenovirus expressing UMAS (Ad-UMAS) was generated using the AdMax^TM system (Hanbio, Shanghai, China) by homologous recombination of pHBAd-MCMV-GFP-UMAS with pHBAd-BHG in HEK-293 cells. The titre of 1011 plaque-forming units (PFU)/mL Ad-UMAS particles was purified by cesium chloride gradient centrifugation and then stored in storage buffer (10 mM Tris, 2 mM MgCl2, 5 % sucrose, pH 8.0) at–80°C. Figure 1 shows the construction of the DNA vaccines (p-MAS and p-UMAS) and adenovirus vaccine (Ad-UMAS).Fig. 1

Bottom Line: DNA vaccines have proved effective in the protection against parasites.Our results indicated that the DNA vaccine had the advantage of inducing a stronger humoral response, whereas the adenovirus-vectored vaccine effectively improved the cellular immune response.Priming vaccination with DNA vaccine and boosting with the recombinant adenovirus vaccine encoding ubiquitin conjugated multi-stage antigens of T. gondii was proved to be a potential strategy against the infection of type I and type II parasite.

View Article: PubMed Central - PubMed

Affiliation: Department of Human Parasitology, Shandong University, School of Medicine, No. 44 Wenhuaxi Road, Jinan, Shandong, 250012, PR China. yinhuiquan521@126.com.

ABSTRACT

Background: Toxoplasma gondii is a widely prevalent intracellular parasite which infects almost all warm-blooded animals including humans and causes serious zoonotic toxoplasmosis. DNA vaccines have proved effective in the protection against parasites. However, the problems of weak immunity and inefficient delivery of DNA vaccine remain major issues. Therefore, comprehensive antigens derived from all stages of the parasite, effective adjuvants and delivery systems should be considered in the vaccine construction.

Methods: SAG3101-144,ROP18347-396, MIC6288-347, GRA7182-224, MAG158-125, BAG1156-211 and SPA142-200, derived from antigens in tachyzoite, bradyzoite and sporozoite stages of T. gondii were screened based on CD8(+) T cell epitope binding affinity to HLA and H-2. We constructed a recombinant DNA vaccine and an adenovirus vaccine encoding multi-stage antigen of T. gondii linked to ubiquitin molecules and vaccinated BALB/c mice with different strategies. Antibodies, cytokines, splenocytes proliferation, as well as the percentage of CD4(+) and CD8(+) T cells in immunized mouse were analyzed by the Enzyme-Linked Immunosorbent Assays (ELISA), Flow Cytometry (FCM). Protective efficacy was evaluated by challenging immunized mice with type I and type II parasite.

Results: Our results indicated that the DNA vaccine had the advantage of inducing a stronger humoral response, whereas the adenovirus-vectored vaccine effectively improved the cellular immune response. Priming with DNA vaccine and boosting with adenovirus-vectored vaccine induced Th1-type immune responses with highest levels of IgG2a and secretion of cytokines IL-2 and IFN-γ. Effective protection against type I and type II parasite with an increase in survival rate and a decrease in brain cyst burden was achieved in immunized mice.

Conclusions: Priming vaccination with DNA vaccine and boosting with the recombinant adenovirus vaccine encoding ubiquitin conjugated multi-stage antigens of T. gondii was proved to be a potential strategy against the infection of type I and type II parasite.

No MeSH data available.


Related in: MedlinePlus