Towards Innovative Design and Application of Recombinant
            <i>Eimeria</i>
            as a Vaccine Vector

Tang, Xinming; Liu, Xianyong; Suo, Xun

doi:10.1128/iai.00861-19

Cited by 12 publications

(6 citation statements)

References 61 publications

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“…Over the past two decades, despite the successful development of a transient and stable transfection platform in Eimeria parasites, their incapacity to complete the entire life cycle and continuously culture in vitro poses obstacles for their genetic manipulation ( 37 ). Presently, the selection system used to enhance the fluorescent rate of transgenic parasites primarily involves a combination of FACS and drug selection in vivo , owing to the low transfection efficiency ( 9 ).…”

Section: Discussionmentioning

confidence: 99%

Oral delivery of Eimeria acervulina transfected sequentially with two copies of the VP2 gene induces immunity against infectious bursal disease virus in chickens

Guo,

Yu,

Suo

et al. 2024

Front. Vet. Sci.

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Chicken coccidiosis caused by Eimeria spp. can occur on almost all poultry farms, causing huge economic losses to the industry. Genetically manipulated Eimeria parasites as a vaccine vector to deliver viral antigens have been reported. In our preliminary study, transgenic E. acervulina expressing a VP2 gene (Ea-VP2) of the infectious bursal disease virus (IBDV) demonstrated partial protection against IBDV infection. To enhance immune responses, we aimed to increase the VP2 gene copy number in transgenic E. acervulina. In this study, we used a novel plasmid vector carrying a VP2 gene fused with three flag tags and a red fluorescent reporter gene (mCherry). The vector was introduced into Ea-VP2 sporozoites through nucleofection, leading to the generation of Ea-2VP2. Subsequent analysis revealed a notable escalation in the fluorescent rate, increasing from 0.11 to 95.1% following four consecutive passages facilitated by fluorescent-activated cell sorting. Verification via PCR, Western blot, and immunofluorescence confirmed the successful construction of the Ea-2VP2 population. Despite lower fecundity compared to wild-type E. acervulina, Ea-2VP2 maintained immunogenicity. Our research effectively created a transgenic E. acervulina strain transfected sequentially with two copies of the VP2 gene from IBDV. This modification resulted in an increased humoral immune response after primary immunization in chickens. Additionally, it demonstrated a degree of protection within the bursa against IBDV infection. Future studies will focus on further enhancing immune response levels.

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Section: Discussionmentioning

confidence: 99%

Oral delivery of Eimeria acervulina transfected sequentially with two copies of the VP2 gene induces immunity against infectious bursal disease virus in chickens

Guo,

Yu,

Suo

et al. 2024

Front. Vet. Sci.

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“…This side effect of reducing average daily gain (g) was important for growing broiler chickens and limits its widespread use in the poultry industry [32], causing great economic losses in the poultry industry. Novel Eimeria formulated with subunits or nucleic acids vaccines have many advantages; some issues limit the recombinant Eimeria, target antigen selection, heterologous antigen expression level, the strength and type of immune responses, and protection against heterologous pathogens [33,34]. As much work still remains to done before the development of recombinant vaccines becomes a reality, it is likely that reliance upon live, attenuated vaccines will increase in years to come [11].…”

Section: Discussionmentioning

confidence: 99%

Lactococcus lactis NZ3900/pNZ8149‐IL‐4‐IL‐2 as an adjuvant to reduce vaccine dose in chicken coccidia live mixed vaccine

Zhang

Fan

et al. 2023

Animal Research and One Health

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Although live mixed coccidiosis vaccines are widely used for Eimeria control, the side effects (e.g., damaging the intestinal mucosa and decreasing their weight gain) are not negligible. This study aimed to filter out the lowest dose of coccidiosis mixed vaccine with Lactococcus lactis (L. lactis) NZ3900/pNZ8149‐IL‐4‐IL‐2 (RLIL4/2) coimmunization against Eimeria reduces the side effects for Eimeria vaccination. Chickens oral 1.0 ×, 0.9 ×, 0.8 ×, 0.6 ×, 0.4 × dose of live vaccine, and RLIL4/2 with 1.0 × 109 CFU/chicken, respectively. The blank control group, challenge‐only group and 1.0 × dose vaccine‐only group are used as references. The survival rate, weight gain, bloody stool, intestinal lesions scores, oocyst output oocysts per gramme (OPG), and anticoccidial index (ACI) were detected by challenging with virulent coccidiosis. All the results indicated that the weight gain during the immune period or bloody stool/intestinal lesions scores/OPG during the challenge period were oppositely correlated with vaccine doses. The survival rate, weight gain, and ACI were positively correlated with vaccine doses during the challenge period. The ACI of 0.6 × the dose group was 167.69, which was considered a moderate efficiency. The weight gain, ACI of 1.0 ×, 0.9 × and 0.8 × dose groups were higher or intestinal lesions were lower than 1.0 × dose vaccine‐only group (p < 0.05), respectively. In conclusion, the coimmunization of RLIL4/2 with Eimeria mixed vaccine could reduce 40% of the dose and the side effects (e.g., weight gain and intestinal lesions) from live mixed coccidiosis vaccine.

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“…Because Eimeria spp. cannot complete their lifecycle or continuously culture in vitro [ 27 ], the genetic manipulation must be performed in vivo and the inoculation route is an essential step to obtain transgenic parasites. Depending on the parasitic site, different inoculation strategies are adopted.…”

Section: Discussionmentioning

confidence: 99%

Establishment of Recombinant Eimeria acervulina Expressing Multi-Copies M2e Derived from Avian Influenza Virus H9N2

et al. 2021

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The potential of Eimeria parasites as live vaccine vectors has been reported with successful genetic manipulation on several species like E. tenella, E. mitis and E. necatrix. Among seven Eimeria species infecting chickens, E. acervulina is a highly prevalent, moderately pathogenic species. Thus, it is valuable for the study of transfection and for use as a potential as vaccine vector. In this study, a plasmid containing expression cassette with enhanced yellow fluorescent protein (EYFP), red fluorescent protein (RFP) and 12 copies of extracellular domain of H9N2 avian influenza virus M2 (M2e) protein was used for the transfection. Nucleofected sporozoites were inoculated into birds through wing vein. Recombinant E. acervulina oocysts with 0.1% EYFP+ and RFP+ populations were collected from the feces of the inoculated birds. The fluorescent rate of transgenic parasites reached over 95% after nine successive propagations with a pyrimethamine selection in vivo and fluorescent-activated cell sorting (FACS) of progeny oocysts. The expression of M2e in the transgenic parasites (EaM2e) was confirmed by Western blot and its cytoplasm localization in sporozoites was displayed by an indirect immunofluorescent assay (IFA). Meanwhile, we found that the fecundity of EaM2e was equivalent to that of wild type E. acervulina (EaWT). Taken together, the stable transfection of E. acervulina was successfully established. Future studies will focus on whether transgenic E. acervulina can serve as a live vaccine vector.

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Towards Innovative Design and Application of Recombinant Eimeria as a Vaccine Vector

Cited by 12 publications

References 61 publications

Oral delivery of Eimeria acervulina transfected sequentially with two copies of the VP2 gene induces immunity against infectious bursal disease virus in chickens

Oral delivery of Eimeria acervulina transfected sequentially with two copies of the VP2 gene induces immunity against infectious bursal disease virus in chickens

Lactococcus lactis NZ3900/pNZ8149‐IL‐4‐IL‐2 as an adjuvant to reduce vaccine dose in chicken coccidia live mixed vaccine

Establishment of Recombinant Eimeria acervulina Expressing Multi-Copies M2e Derived from Avian Influenza Virus H9N2

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