Neospora caninum antigens displaying virus-like particles as a bivalent vaccine candidate against neosporosis

Xu, Jian; Hiramatsu, Rikito; Suhaimi, Hamizah; Kato, Tatsuya; Fujimoto, Akari; Tokiwa, Toshihiro; Ike, Kazunori; Park, Enoch Y.

doi:10.1016/j.vaccine.2019.09.002

Cited by 8 publications

(5 citation statements)

References 41 publications

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“…While a plethora of potential neosporosis vaccine candidates have been evaluated in recent years [35,36], NcSAG1 was chosen as target antigen since it represents a highly immunodominant, surface protein of N. caninum tachyzoites, which is implicated in the host cell invasion process [37,38]. NcSAG1 applied as DNA vaccine and as recombinant antigen [39][40][41][42] had been shown earlier to confer partial protection against cerebral infection in non-pregnant mice.…”

Section: Discussionmentioning

confidence: 99%

A Listeria monocytogenes-Based Vaccine Formulation Reduces Vertical Transmission and Leads to Enhanced Pup Survival in a Pregnant Neosporosis Mouse Model

et al. 2021

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The apicomplexan parasite Neospora caninum is the worldwide leading cause of abortion and stillbirth in cattle. An attenuated mutant Listeria monocytogenes strain (Lm3Dx) was engineered by deleting the virulence genes actA, inlA, and inlB in order to avoid systemic infection and to target the vector to antigen-presenting cells (APCs). Insertion of sag1, coding for the major surface protein NcSAG1 of N. caninum, yielded the vaccine strain Lm3Dx_NcSAG1. The efficacy of Lm3Dx_NcSAG1 was assessed by inoculating 1 × 105, 1 × 106, or 1 × 107 CFU of Lm3Dx_NcSAG1 into female BALB/c mice by intramuscular injection three times at two-week intervals, and subsequent challenge with 1 × 105N. caninum tachyzoites of the highly virulent NcSpain-7 strain on day 7 of pregnancy. Dose-dependent protective effects were seen, with a postnatal offspring survival rate of 67% in the group treated with 1 × 107 CFU of Lm3Dx_NcSAG1 compared to 5% survival in the non-vaccinated control group. At euthanasia (25 days post-partum), IgG antibody titers were significantly decreased in the groups receiving the two higher doses and cytokines recall responses in splenocyte culture supernatants (IFN-γ, IL-4, and IL-10) were increased in the vaccinated groups. Thus, Lm3Dx_NcSAG1 induces immune-protective effects associated with a balanced Th1/Th2 response in a pregnant neosporosis mouse model and should be further assessed in ruminant models.

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Section: Discussionmentioning

confidence: 99%

A Listeria monocytogenes-Based Vaccine Formulation Reduces Vertical Transmission and Leads to Enhanced Pup Survival in a Pregnant Neosporosis Mouse Model

et al. 2021

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“…This result indicates that the 3C proteinase was functionally expressed in silkworm-BEVS. Although we did not continue to purify the GST-3C, we can expect a satisfactory amount of 3C protein from silkworm larvae since scaleup expression can be simply performed by increasing the sample size [ 29 , 35 , 37 ].…”

Section: Resultsmentioning

confidence: 99%

In vivo enzymatic digestion of HRV 3C protease cleavage sites-containing proteins produced in a silkworm-baculovirus expression system

Nakanishi

Kato

et al. 2022

Bioscience Reports

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Baculovirus expression vector system (BEVS) has been recognized as a potent protein expression system in engineering valuable enzymes and vaccines. Various fusion tags facilitate protein purification, leaving the potential risk to influence the target protein’s biological activity negatively. It is of great interest to consider removing the additional tags using site-specific proteases, such as human rhinoviruses (HRV) 3C protease. The current study validated the cleavage activity of 3C protease in Escherichia coli and silkworm-BEVS systems by mixing the cell or fat body lysates of 3C protein and 3C site containing target protein in vitro. Further verification has been performed in the fat body lysate from co-expression of both constructs, showing remarkable cleavage efficiency in vivo silkworm larvae. We also achieved the glutathione-S-transferase (GST) tag-cleaved product of the VP15 protein from the White spot syndrome virus after purification, suggesting that we successfully established a coinfection-based recognition-and-reaction BEVS platform for the tag-free protein engineering.

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“…For instance, NcSAG1 is constitutively expressed on the tachyzoite surface and is likely to mediate the initial contact between the parasite and the host cell surface membrane. NcSAG1 has been previously assessed as a vaccine candidate by employing different expression systems such as DNA-vaccines and recombinant antigens [ 21 ], the integration of a recombinant antigen into immune-stimulating complexes (ISCOMs) [ 44 ] and expression in silkworm larvae ( Bombyx mori ) using nucleopolyhedrovirus (BmNPV) bacmid expression systems [ 45 ] or Rous sarcoma virus-like particles (RSV-LP) displaying both NcSAG1 and NcSRS2 [ 46 ]. NcROP2 has also been shown earlier to represent a promising vaccine candidate, either alone or in combination with other recombinant antigens [ 38 ].…”

Section: Discussionmentioning

confidence: 99%

Immunization with a Multivalent Listeria monocytogenes Vaccine Leads to a Strong Reduction in Vertical Transmission and Cerebral Parasite Burden in Pregnant and Non-Pregnant Mice Infected with Neospora caninum

et al. 2023

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Neospora caninum is an apicomplexan parasite that causes abortion and stillbirth in cattle. We employed the pregnant neosporosis mouse model to investigate the efficacy of a modified version of the attenuated Listeria monocytogenes vaccine vector Lm3Dx_NcSAG1, which expresses the major N. caninum surface antigen SAG1. Multivalent vaccines were generated by the insertion of gra7 and/or rop2 genes into Lm3Dx_NcSAG1, resulting in the double mutants, Lm3Dx_NcSAG1_NcGRA7 and Lm3Dx_NcSAG1_NcROP2, and the triple mutant, Lm3Dx_NcSAG1_NcGRA7_NcROP2. Six experimental groups of female BALB/c mice were inoculated intramuscularly three times at two-week intervals with 1 × 107 CFU of the respective vaccine strains. Seven days post-mating, mice were challenged by the subcutaneous injection of 1 × 105N. caninum NcSpain-7 tachyzoites. Non-pregnant mice, dams and their offspring were observed daily until day 25 post-partum. Immunization with Lm3Dx_NcSAG1 and Lm3Dx_NcSAG1_NcGRA7_NcROP2 resulted in 70% postnatal pup survival, whereas only 50% and 58% of pups survived in the double mutant-vaccinated groups. Almost all pups had died at the end of the experiment in the infection control. The triple mutant was the most promising vaccine candidate, providing the highest rate of protection against vertical transmission (65%) and CNS infection. Overall, integrating multiple antigens into Lm3Dx_SAG1 resulted in lower vertical transmission and enhanced protection against cerebral infection in dams and in non-pregnant mice.

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Neospora caninum antigens displaying virus-like particles as a bivalent vaccine candidate against neosporosis

Cited by 8 publications

References 41 publications

A Listeria monocytogenes-Based Vaccine Formulation Reduces Vertical Transmission and Leads to Enhanced Pup Survival in a Pregnant Neosporosis Mouse Model

A Listeria monocytogenes-Based Vaccine Formulation Reduces Vertical Transmission and Leads to Enhanced Pup Survival in a Pregnant Neosporosis Mouse Model

In vivo enzymatic digestion of HRV 3C protease cleavage sites-containing proteins produced in a silkworm-baculovirus expression system

Immunization with a Multivalent Listeria monocytogenes Vaccine Leads to a Strong Reduction in Vertical Transmission and Cerebral Parasite Burden in Pregnant and Non-Pregnant Mice Infected with Neospora caninum

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