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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

Proliferation responses against V. harveyi in immunized grouper. Four groups of grouper were intraperitoneally immunized twice (↑) with PLG-PLE/rGAPDH microparticles (■), PLG-rGAPDH microparticles (●), rGAPDH alone (□) or PBS (○). After immunization, V. harveyi lysate-stimulated head kidney lymphocytes were prepared from three fish per group every three weeks, and their subsequent proliferation responses were analyzed and expressed as stimulation index (SI) values. Results were presented as the mean of SI values ± SD. * p < 0.05 when comparing the PLG-PLE/rGAPDH group to the PLG-rGAPDH group.
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vaccines-02-00380-f006: Proliferation responses against V. harveyi in immunized grouper. Four groups of grouper were intraperitoneally immunized twice (↑) with PLG-PLE/rGAPDH microparticles (■), PLG-rGAPDH microparticles (●), rGAPDH alone (□) or PBS (○). After immunization, V. harveyi lysate-stimulated head kidney lymphocytes were prepared from three fish per group every three weeks, and their subsequent proliferation responses were analyzed and expressed as stimulation index (SI) values. Results were presented as the mean of SI values ± SD. * p < 0.05 when comparing the PLG-PLE/rGAPDH group to the PLG-rGAPDH group.

Mentions: Every three weeks, head kidney lymphocytes stimulated with V. harveyi lysate were prepared from different groups of fish, and their subsequent proliferation responses were analyzed and expressed as SI values (Figure 6). Both PLG-PLE/rGAPDH and PLG-rGAPDH microparticles, respectively, gave rise to sustained lymphocyte proliferation for twelve weeks. Three weeks after boosting (the sixth week), PLG-PLE/rGAPDH microparticles elicited significantly higher SI values (p < 0.05, nested design) than PLG-rGAPDH microparticles did (Figure 6). However, administration with soluble rGAPDH alone or PBS induced little lymphocyte proliferation in grouper (Figure 6). The synthetic PLE peptide, therefore, enhanced grouper anti-V. harveyi lymphocyte proliferation elicited by immunization with PLG-encapsulated rGAPDH protein.


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)

Proliferation responses against V. harveyi in immunized grouper. Four groups of grouper were intraperitoneally immunized twice (↑) with PLG-PLE/rGAPDH microparticles (■), PLG-rGAPDH microparticles (●), rGAPDH alone (□) or PBS (○). After immunization, V. harveyi lysate-stimulated head kidney lymphocytes were prepared from three fish per group every three weeks, and their subsequent proliferation responses were analyzed and expressed as stimulation index (SI) values. Results were presented as the mean of SI values ± SD. * 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-f006: Proliferation responses against V. harveyi in immunized grouper. Four groups of grouper were intraperitoneally immunized twice (↑) with PLG-PLE/rGAPDH microparticles (■), PLG-rGAPDH microparticles (●), rGAPDH alone (□) or PBS (○). After immunization, V. harveyi lysate-stimulated head kidney lymphocytes were prepared from three fish per group every three weeks, and their subsequent proliferation responses were analyzed and expressed as stimulation index (SI) values. Results were presented as the mean of SI values ± SD. * p < 0.05 when comparing the PLG-PLE/rGAPDH group to the PLG-rGAPDH group.
Mentions: Every three weeks, head kidney lymphocytes stimulated with V. harveyi lysate were prepared from different groups of fish, and their subsequent proliferation responses were analyzed and expressed as SI values (Figure 6). Both PLG-PLE/rGAPDH and PLG-rGAPDH microparticles, respectively, gave rise to sustained lymphocyte proliferation for twelve weeks. Three weeks after boosting (the sixth week), PLG-PLE/rGAPDH microparticles elicited significantly higher SI values (p < 0.05, nested design) than PLG-rGAPDH microparticles did (Figure 6). However, administration with soluble rGAPDH alone or PBS induced little lymphocyte proliferation in grouper (Figure 6). The synthetic PLE peptide, therefore, enhanced grouper anti-V. harveyi lymphocyte proliferation elicited by immunization with PLG-encapsulated rGAPDH protein.

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