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Pleurocidin Peptide Enhances Grouper Anti-Vibrio harveyi Immunity Elicited by Poly(lactide-co-glycolide)-Encapsulated Recombinant Glyceraldehyde-3-phosphate Dehydrogenase.

Chuang SC, Huang WL, Kau SW, Yang YP, Yang CD - Vaccines (Basel) (2014)

Bottom Line: The resulting PLG-encapsulated PLE plus rGAPDH (PLG-PLE/rGAPDH) microparticles, 3.21-6.27 μm in diameter, showed 72%-83% entrapment efficiency and durably released both PLE and rGAPDH for a long 30-day period.After an experimental challenge of V. harveyi, PLG-PLE/rGAPDH microparticles conferred a high survival rate (85%), which was significantly higher (p < 0.05, chi-square test) than that induced by PLG-rGAPDH microparticles (67%).In conclusion, PLE peptide exhibits an efficacious adjuvant effect to elicit not only improved immunity, but also enhanced protection against V. harveyi in grouper induced by rGAPDH protein encapsulated in PLG microparticles.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, College of Medicine, Kaohsiung Medical University, No. 100, Shih-Chuan 1st Road, Kaohsiung 807, Taiwan. f86225016@ntu.edu.tw.

ABSTRACT
Outer membrane proteins, such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH), are considered immunodominant antigens for eliciting protective immunity against Vibrio harveyi, the main etiological agent of vibriosis in fish. Cationic antimicrobial peptides (AMPs), such as pleurocidin (PLE), play important roles in activating and recruiting immune cells, thereby contributing to subsequent innate and adaptive immune responses. In the present study, we aimed to use PLE peptide as a potent adjuvant to improve the immunogenicity of V. harveyi recombinant GAPDH (rGAPDH). In order to prepare a controlled-release vaccine, PLE peptide and rGAPDH protein were simultaneously encapsulated into polymeric microparticles made from the biodegradable poly(lactide-co-glycolide) (PLG) polymer. The resulting PLG-encapsulated PLE plus rGAPDH (PLG-PLE/rGAPDH) microparticles, 3.21-6.27 μm in diameter, showed 72%-83% entrapment efficiency and durably released both PLE and rGAPDH for a long 30-day period. Following peritoneal immunization in grouper (Epinephelus coioides), PLG-PLE/rGAPDH microparticles resulted in significantly higher (p < 0.05, nested design) long-lasting GAPDH-specific immunity (serum titers and lymphocyte proliferation) than PLG-encapsulated rGAPDH (PLG-rGAPDH) microparticles. After an experimental challenge of V. harveyi, PLG-PLE/rGAPDH microparticles conferred a high survival rate (85%), which was significantly higher (p < 0.05, chi-square test) than that induced by PLG-rGAPDH microparticles (67%). In conclusion, PLE peptide exhibits an efficacious adjuvant effect to elicit not only improved immunity, but also enhanced protection against V. harveyi in grouper induced by rGAPDH protein encapsulated in PLG microparticles.

No MeSH data available.


Related in: MedlinePlus

Scanning electron micrographs of PLG-encapsulated microparticles. poly(Lactide-co-glycolide) (PLG)-pleurocidin (PLE)/rGAPDH microparticles (A) and PLG-rGAPDH microparticles (B) can be seen as spherical particles with a smooth surface (the bar represents 20 μm).
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vaccines-02-00380-f002: Scanning electron micrographs of PLG-encapsulated microparticles. poly(Lactide-co-glycolide) (PLG)-pleurocidin (PLE)/rGAPDH microparticles (A) and PLG-rGAPDH microparticles (B) can be seen as spherical particles with a smooth surface (the bar represents 20 μm).

Mentions: rGAPDH alone and rGAPDH mixed with synthetic PLE peptide were respectively encapsulated into PLG microparticles. The resulting PLG-rGAPDH and PLG-PLE/rGAPDH microparticles were then analyzed. The morphology of microparticles was first inspected by scanning electron microscopy. Both PLG-PLE/rGAPDH and PLG-rGAPDH microparticles showed a uniform population of spherical particles with a smooth surface (Figure 2). A particle size analyzer was further used to determine the particle size. Different PLG-PLE/rGAPDH preparations showed a mean diameter ranging from 3.21 to 6.27 µm, and their entrapment efficiency for rGAPDH plus PLE ranged from 72% to 83%, with a mean encapsulation efficiency of 77% (Table 1). In each preparation of PLG-PLE/rGAPDH microparticles (10 mg of rGAPDH and 1 mg of PLE were used), therefore, the total amount of 7.7 mg rGAPDH and 0.77 mg PLE was actually encapsulated into PLG microparticles. The cumulative release of rGAPDH and PLE during the course of a 30-day period was 82% of the total protein/peptide load (data not shown). Therefore, 6.31 mg of rGAPDH and 0.63 mg of PLE were released from the original microparticle. In addition, another PLG microparticle vaccine, PLG-rGAPDH microparticles, was also prepared in the present study. As shown in Table 1, the mean diameter of different PLG-rGAPDH batches ranged from 2.84 to 6.02 µm, and the entrapment efficiency for rGAPDH was from 69% to 78%, with a mean encapsulation efficiency of 74%. In each preparation of PLG-rGAPDH microparticles (10 mg of rGAPDH were used), the total amount of 7.4 mg rGAPDH was actually encapsulated into PLG microparticles. The cumulative release of rGAPDH during the course of a 30-day period was 80% of the total protein load (data not shown). Therefore, 5.92 mg of rGAPDH was released from the original microparticle. In order to determine if the prepared microparticles released protein and/or peptide, 18% SDS-PAGE was undertaken to examine released samples collected on different days (Figure 3). Results showed that the in vitro release of rGAPDH plus PLE peptide (Figure 3A) or rGAPDH alone (Figure 3B) from PLG microparticles could be maintained during the course of a long 30-day period.


Pleurocidin Peptide Enhances Grouper Anti-Vibrio harveyi Immunity Elicited by Poly(lactide-co-glycolide)-Encapsulated Recombinant Glyceraldehyde-3-phosphate Dehydrogenase.

Chuang SC, Huang WL, Kau SW, Yang YP, Yang CD - Vaccines (Basel) (2014)

Scanning electron micrographs of PLG-encapsulated microparticles. poly(Lactide-co-glycolide) (PLG)-pleurocidin (PLE)/rGAPDH microparticles (A) and PLG-rGAPDH microparticles (B) can be seen as spherical particles with a smooth surface (the bar represents 20 μm).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494259&req=5

vaccines-02-00380-f002: Scanning electron micrographs of PLG-encapsulated microparticles. poly(Lactide-co-glycolide) (PLG)-pleurocidin (PLE)/rGAPDH microparticles (A) and PLG-rGAPDH microparticles (B) can be seen as spherical particles with a smooth surface (the bar represents 20 μm).
Mentions: rGAPDH alone and rGAPDH mixed with synthetic PLE peptide were respectively encapsulated into PLG microparticles. The resulting PLG-rGAPDH and PLG-PLE/rGAPDH microparticles were then analyzed. The morphology of microparticles was first inspected by scanning electron microscopy. Both PLG-PLE/rGAPDH and PLG-rGAPDH microparticles showed a uniform population of spherical particles with a smooth surface (Figure 2). A particle size analyzer was further used to determine the particle size. Different PLG-PLE/rGAPDH preparations showed a mean diameter ranging from 3.21 to 6.27 µm, and their entrapment efficiency for rGAPDH plus PLE ranged from 72% to 83%, with a mean encapsulation efficiency of 77% (Table 1). In each preparation of PLG-PLE/rGAPDH microparticles (10 mg of rGAPDH and 1 mg of PLE were used), therefore, the total amount of 7.7 mg rGAPDH and 0.77 mg PLE was actually encapsulated into PLG microparticles. The cumulative release of rGAPDH and PLE during the course of a 30-day period was 82% of the total protein/peptide load (data not shown). Therefore, 6.31 mg of rGAPDH and 0.63 mg of PLE were released from the original microparticle. In addition, another PLG microparticle vaccine, PLG-rGAPDH microparticles, was also prepared in the present study. As shown in Table 1, the mean diameter of different PLG-rGAPDH batches ranged from 2.84 to 6.02 µm, and the entrapment efficiency for rGAPDH was from 69% to 78%, with a mean encapsulation efficiency of 74%. In each preparation of PLG-rGAPDH microparticles (10 mg of rGAPDH were used), the total amount of 7.4 mg rGAPDH was actually encapsulated into PLG microparticles. The cumulative release of rGAPDH during the course of a 30-day period was 80% of the total protein load (data not shown). Therefore, 5.92 mg of rGAPDH was released from the original microparticle. In order to determine if the prepared microparticles released protein and/or peptide, 18% SDS-PAGE was undertaken to examine released samples collected on different days (Figure 3). Results showed that the in vitro release of rGAPDH plus PLE peptide (Figure 3A) or rGAPDH alone (Figure 3B) from PLG microparticles could be maintained during the course of a long 30-day period.

Bottom Line: The resulting PLG-encapsulated PLE plus rGAPDH (PLG-PLE/rGAPDH) microparticles, 3.21-6.27 μm in diameter, showed 72%-83% entrapment efficiency and durably released both PLE and rGAPDH for a long 30-day period.After an experimental challenge of V. harveyi, PLG-PLE/rGAPDH microparticles conferred a high survival rate (85%), which was significantly higher (p < 0.05, chi-square test) than that induced by PLG-rGAPDH microparticles (67%).In conclusion, PLE peptide exhibits an efficacious adjuvant effect to elicit not only improved immunity, but also enhanced protection against V. harveyi in grouper induced by rGAPDH protein encapsulated in PLG microparticles.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, College of Medicine, Kaohsiung Medical University, No. 100, Shih-Chuan 1st Road, Kaohsiung 807, Taiwan. f86225016@ntu.edu.tw.

ABSTRACT
Outer membrane proteins, such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH), are considered immunodominant antigens for eliciting protective immunity against Vibrio harveyi, the main etiological agent of vibriosis in fish. Cationic antimicrobial peptides (AMPs), such as pleurocidin (PLE), play important roles in activating and recruiting immune cells, thereby contributing to subsequent innate and adaptive immune responses. In the present study, we aimed to use PLE peptide as a potent adjuvant to improve the immunogenicity of V. harveyi recombinant GAPDH (rGAPDH). In order to prepare a controlled-release vaccine, PLE peptide and rGAPDH protein were simultaneously encapsulated into polymeric microparticles made from the biodegradable poly(lactide-co-glycolide) (PLG) polymer. The resulting PLG-encapsulated PLE plus rGAPDH (PLG-PLE/rGAPDH) microparticles, 3.21-6.27 μm in diameter, showed 72%-83% entrapment efficiency and durably released both PLE and rGAPDH for a long 30-day period. Following peritoneal immunization in grouper (Epinephelus coioides), PLG-PLE/rGAPDH microparticles resulted in significantly higher (p < 0.05, nested design) long-lasting GAPDH-specific immunity (serum titers and lymphocyte proliferation) than PLG-encapsulated rGAPDH (PLG-rGAPDH) microparticles. After an experimental challenge of V. harveyi, PLG-PLE/rGAPDH microparticles conferred a high survival rate (85%), which was significantly higher (p < 0.05, chi-square test) than that induced by PLG-rGAPDH microparticles (67%). In conclusion, PLE peptide exhibits an efficacious adjuvant effect to elicit not only improved immunity, but also enhanced protection against V. harveyi in grouper induced by rGAPDH protein encapsulated in PLG microparticles.

No MeSH data available.


Related in: MedlinePlus