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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

Antigenic specificity in immunized grouper sera. Three weeks after boosting, grouper sera were collected to analyze their antigenic specificity. V. harveyi lysate was probed with sera from grouper peritoneally immunized with PLG-PLE/rGAPDH microparticles (Lane 1), PLG-rGAPDH microparticles (Lane 2), rGAPDH alone (Lane 3) or PBS (Lane 4). The V. harveyi-infected grouper serum was also conducted (Lane 5). Standard protein markers (Lane M) are shown on the left.
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vaccines-02-00380-f004: Antigenic specificity in immunized grouper sera. Three weeks after boosting, grouper sera were collected to analyze their antigenic specificity. V. harveyi lysate was probed with sera from grouper peritoneally immunized with PLG-PLE/rGAPDH microparticles (Lane 1), PLG-rGAPDH microparticles (Lane 2), rGAPDH alone (Lane 3) or PBS (Lane 4). The V. harveyi-infected grouper serum was also conducted (Lane 5). Standard protein markers (Lane M) are shown on the left.

Mentions: The ability of PLG microparticles to trigger humoral immunity against V. harveyi in grouper was subsequently evaluated. Western blot studies of fish sera obtained three weeks after boosting showed that both PLG-rGAPDH and PLG-PLE/rGAPDH microparticles resulted in the production of grouper serum antibodies against the native GAPDH protein in V. harveyi lysate (Figure 4, Lanes 1 and 2). However, sera from grouper immunized with rGAPDH alone or PBS did not recognize any proteins in V. harveyi lysate (Figure 4, Lanes 3 and 4). Therefore, intraperitoneal immunization with rGAPDH in grouper could elicit a specific serum response to the native GAPDH protein in V. harveyi lysate only when rGAPDH protein was encapsulated with the PLG polymer, but not in its soluble form. In addition, every three weeks, the specific anti-V. harveyi serum titers in grouper were determined by ELISA (Figure 5). Intraperitoneal immunization with PLG-PLE/rGAPDH or PLG-rGAPDH microparticles was able to durably keep high serum titers, up till the 12th week. More importantly, three weeks after boosting (the sixth week), grouper serum titers induced by PLG-PLE/rGAPDH microparticles were significantly higher (p < 0.05, nested design) than those of PLG-rGAPDH microparticles (Figure 5). However, grouper immunized with soluble rGAPDH alone or PBS displayed little, if any, anti-V. harveyi serum titers (Figure 5). Therefore, the presence of PLE peptide improved the grouper anti-V. harveyi serum response induced by rGAPDH encapsulated in PLG microparticles.


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)

Antigenic specificity in immunized grouper sera. Three weeks after boosting, grouper sera were collected to analyze their antigenic specificity. V. harveyi lysate was probed with sera from grouper peritoneally immunized with PLG-PLE/rGAPDH microparticles (Lane 1), PLG-rGAPDH microparticles (Lane 2), rGAPDH alone (Lane 3) or PBS (Lane 4). The V. harveyi-infected grouper serum was also conducted (Lane 5). Standard protein markers (Lane M) are shown on the left.
© Copyright Policy
Related In: Results  -  Collection

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

vaccines-02-00380-f004: Antigenic specificity in immunized grouper sera. Three weeks after boosting, grouper sera were collected to analyze their antigenic specificity. V. harveyi lysate was probed with sera from grouper peritoneally immunized with PLG-PLE/rGAPDH microparticles (Lane 1), PLG-rGAPDH microparticles (Lane 2), rGAPDH alone (Lane 3) or PBS (Lane 4). The V. harveyi-infected grouper serum was also conducted (Lane 5). Standard protein markers (Lane M) are shown on the left.
Mentions: The ability of PLG microparticles to trigger humoral immunity against V. harveyi in grouper was subsequently evaluated. Western blot studies of fish sera obtained three weeks after boosting showed that both PLG-rGAPDH and PLG-PLE/rGAPDH microparticles resulted in the production of grouper serum antibodies against the native GAPDH protein in V. harveyi lysate (Figure 4, Lanes 1 and 2). However, sera from grouper immunized with rGAPDH alone or PBS did not recognize any proteins in V. harveyi lysate (Figure 4, Lanes 3 and 4). Therefore, intraperitoneal immunization with rGAPDH in grouper could elicit a specific serum response to the native GAPDH protein in V. harveyi lysate only when rGAPDH protein was encapsulated with the PLG polymer, but not in its soluble form. In addition, every three weeks, the specific anti-V. harveyi serum titers in grouper were determined by ELISA (Figure 5). Intraperitoneal immunization with PLG-PLE/rGAPDH or PLG-rGAPDH microparticles was able to durably keep high serum titers, up till the 12th week. More importantly, three weeks after boosting (the sixth week), grouper serum titers induced by PLG-PLE/rGAPDH microparticles were significantly higher (p < 0.05, nested design) than those of PLG-rGAPDH microparticles (Figure 5). However, grouper immunized with soluble rGAPDH alone or PBS displayed little, if any, anti-V. harveyi serum titers (Figure 5). Therefore, the presence of PLE peptide improved the grouper anti-V. harveyi serum response induced by rGAPDH encapsulated in PLG microparticles.

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