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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

Survival of immunized grouper after a lethal V. harveyi challenge. Grouper were intraperitoneally immunized twice with PLG-PLE/rGAPDH microparticles (■), PLG-rGAPDH microparticles (●), rGAPDH alone (□) or PBS (○). Nine weeks after boosting, four groups of 60 groupers each were intraperitoneally infected with 6 × 106 CFU of V. harveyi (BCRC13812 strain). Fish were observed daily for an additional month (28 days), and the final survival rates were calculated. * p < 0.05 when comparing the PLG-PLE/rGAPDH group to the PLG-rGAPDH group.
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vaccines-02-00380-f007: Survival of immunized grouper after a lethal V. harveyi challenge. Grouper were intraperitoneally immunized twice with PLG-PLE/rGAPDH microparticles (■), PLG-rGAPDH microparticles (●), rGAPDH alone (□) or PBS (○). Nine weeks after boosting, four groups of 60 groupers each were intraperitoneally infected with 6 × 106 CFU of V. harveyi (BCRC13812 strain). Fish were observed daily for an additional month (28 days), and the final survival rates were calculated. * p < 0.05 when comparing the PLG-PLE/rGAPDH group to the PLG-rGAPDH group.

Mentions: We then determined whether PLG microparticle vaccines could confer effective protection in grouper. Nine weeks after boosting (the 12th week), all groups of 60 groupers each were intraperitoneally challenged with 6 × 106 CFU of V. harveyi (BCRC13812 strain). Fish were observed daily for an additional month (28 days) and the survival rates were recorded (Figure 7). All fish administrated with soluble rGAPDH alone or PBS died within 16 days after challenge and displayed no protection against the experimental challenge. Forty fish immunized with PLG-rGAPDH microparticles survived during the challenge study and showed a protection rate of 67%. However, in the group of fish immunized with PLG-PLE/rGAPDH microparticles, only nine fish died during the challenge study. Thus, intraperitoneal immunization with PLG-PLE/rGAPDH microparticles in grouper resulted in the highest survival rate (85%), which was significantly higher (p < 0.05, chi-square test) than that of the PLG-rGAPDH group (67%) (Figure 7). Therefore, the addition of the peptide adjuvant, PLE, into the vaccine formulation provided a substantial advance in protection against V. harveyi.


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)

Survival of immunized grouper after a lethal V. harveyi challenge. Grouper were intraperitoneally immunized twice with PLG-PLE/rGAPDH microparticles (■), PLG-rGAPDH microparticles (●), rGAPDH alone (□) or PBS (○). Nine weeks after boosting, four groups of 60 groupers each were intraperitoneally infected with 6 × 106 CFU of V. harveyi (BCRC13812 strain). Fish were observed daily for an additional month (28 days), and the final survival rates were calculated. * p < 0.05 when comparing the PLG-PLE/rGAPDH group to the PLG-rGAPDH group.
© Copyright Policy
Related In: Results  -  Collection

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

vaccines-02-00380-f007: Survival of immunized grouper after a lethal V. harveyi challenge. Grouper were intraperitoneally immunized twice with PLG-PLE/rGAPDH microparticles (■), PLG-rGAPDH microparticles (●), rGAPDH alone (□) or PBS (○). Nine weeks after boosting, four groups of 60 groupers each were intraperitoneally infected with 6 × 106 CFU of V. harveyi (BCRC13812 strain). Fish were observed daily for an additional month (28 days), and the final survival rates were calculated. * p < 0.05 when comparing the PLG-PLE/rGAPDH group to the PLG-rGAPDH group.
Mentions: We then determined whether PLG microparticle vaccines could confer effective protection in grouper. Nine weeks after boosting (the 12th week), all groups of 60 groupers each were intraperitoneally challenged with 6 × 106 CFU of V. harveyi (BCRC13812 strain). Fish were observed daily for an additional month (28 days) and the survival rates were recorded (Figure 7). All fish administrated with soluble rGAPDH alone or PBS died within 16 days after challenge and displayed no protection against the experimental challenge. Forty fish immunized with PLG-rGAPDH microparticles survived during the challenge study and showed a protection rate of 67%. However, in the group of fish immunized with PLG-PLE/rGAPDH microparticles, only nine fish died during the challenge study. Thus, intraperitoneal immunization with PLG-PLE/rGAPDH microparticles in grouper resulted in the highest survival rate (85%), which was significantly higher (p < 0.05, chi-square test) than that of the PLG-rGAPDH group (67%) (Figure 7). Therefore, the addition of the peptide adjuvant, PLE, into the vaccine formulation provided a substantial advance in protection against V. harveyi.

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