Protective Immunity against Vibrio harveyi in Grouper Induced by Single Vaccination with Poly (Lactide-co-glycolide) Microparticles Releasing Pleurocidin Peptide and Recombinant Glyceraldehyde-3-phosphate Dehydrogenase
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacterial Culture
2.2. Preparation of PLG-PLE/rGAPDH MPs
2.3. Inactivated V. harveyi Bacteria
2.4. Bacterial Lysate
2.5. Animals
2.6. Immunization
2.7. Serum Assay
2.8. Lymphocyte Proliferation Assay
2.9. Quantitative Real-Time Reverse Transcriptase-PCR (qRT-PCR)
2.10. Bacterial Challenge
2.11. Statistical Analysis
3. Results
3.1. Antigenic Specificity of Grouper Antisera Following Single Immunization
3.2. Sustained High Serum Titers Raised by Single Vaccination with PLG-PLE/rGAPDH MPs
3.3. Robust Lymphocyte Proliferation Produced by Single Vaccination with PLG-PLE/rGAPDH MPs
3.4. High TNF-α Production Following Single Vaccination with PLG-PLE/rGAPDH MPs
3.5. Strong Protection against V. harveyi in Grouper Induced by Single Vaccination with PLG-PLE/rGAPDH MPs
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Primer Name | Nucleotide Sequence (5′ → 3′) | Amplification Size (bp) | Accession Number/Reference |
---|---|---|---|
TNF-α (F) | GTGTCCTGCTGTTTGCTTGGTA | 207 | FJ009049/[29] |
TNF-α (R) | CAGTGTCCGACTTGATTAGTGCTT | ||
β actin (F) | TGCTGTCCCTGTATGCCTCT | 225 | AY510710/[30] |
β actin (R) | CCTTGATGTCACGCACGAT |
Group a | TNF-α (Fold Change) b | |
---|---|---|
Lysate | Con A | |
PLG-PLE/rGAPDH MPs | 12.4 ± 1.5 c | 2.7 ± 0.9 f |
Inactivated bacteria (ISA 763 AVG) | 3.2 ± 0.5 d | 2.8 ± 0.6 f |
rGAPDH | 1.4 ± 0.3 e | 2.7 ± 0.7 f |
PLE | 1.2 ± 0.4 e | 2.6 ± 0.5 f |
PLG empty MPs | 1.1 ± 0.2 e | 2.5 ± 0.7 f |
PBS | 0.9 ± 0.3 e | 2.4 ± 0.5 f |
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Liu, S.-P.; Chuang, S.-C.; Yang, C.-D. Protective Immunity against Vibrio harveyi in Grouper Induced by Single Vaccination with Poly (Lactide-co-glycolide) Microparticles Releasing Pleurocidin Peptide and Recombinant Glyceraldehyde-3-phosphate Dehydrogenase. Vaccines 2020, 8, 33. https://doi.org/10.3390/vaccines8010033
Liu S-P, Chuang S-C, Yang C-D. Protective Immunity against Vibrio harveyi in Grouper Induced by Single Vaccination with Poly (Lactide-co-glycolide) Microparticles Releasing Pleurocidin Peptide and Recombinant Glyceraldehyde-3-phosphate Dehydrogenase. Vaccines. 2020; 8(1):33. https://doi.org/10.3390/vaccines8010033
Chicago/Turabian StyleLiu, Shang-Pin, Shu-Chun Chuang, and Chung-Da Yang. 2020. "Protective Immunity against Vibrio harveyi in Grouper Induced by Single Vaccination with Poly (Lactide-co-glycolide) Microparticles Releasing Pleurocidin Peptide and Recombinant Glyceraldehyde-3-phosphate Dehydrogenase" Vaccines 8, no. 1: 33. https://doi.org/10.3390/vaccines8010033