Dataset Information

Oral Vaccination With Recombinant Pichia pastoris Expressing Iridovirus Major Capsid Protein Elicits Protective Immunity in Largemouth Bass (Micropterus salmoides).

ABSTRACT: Largemouth bass iridovirus (LMBV) can cause high mortality and lead to heavy economic loss in the cultivation of largemouth bass, but there was no effective treatment. Here, the present study constructed a recombinant Pichia pastoris expressing LMBV major capsid protein (MCPD). The recombinant GS115-pW317-MCPD was then used to immunize largemouth bass via oral administration, and mucosal immune response mediated by immunoglobulins (Igs) was measured after oral immunization. Serum antibody levels were measured by ELISA, neutralizing antibody titers were determined by serum neutralization test (SNT), antigen presentation-related gene expressions were detected by RT-PCR, and the histopathological characteristics of immunized fish were assessed after challenging with 0.1 ml 10^7.19 TCID₅₀/ml LMBV. The relative percentage survival (RPS) was also determined. Our results showed that the serum antibody titers of immunized fish were significantly higher than that of control groups (P < 0.05). IgT and IgM expressions in gut were increased significantly after vaccination with GS115-pW317-MCPD; however, much stronger response in gut was observed as compared with gill. The expression levels of major histocompatibility complex (MHC) II, CD8, and T-cell receptor (TCR) were significantly elevated in GS115-pW317-MCPD group (P < 0.05), while CD4 and MHC I transcription levels remained unchanged after oral immunization (P > 0.05). The RPS of fish orally immunized with 1.0 × 10⁸ CFU/g GS115-pW317-MCPD was reached up to 41.6% after challenge with 0.1 ml 10^9.46 TCID₅₀/ml LMBV. Moreover, orally immunizing with GS115-pW317-MCPD can relieve the pathological damage caused by LMBV. Therefore, GS115-pW317-MCPD showed a promising potential against LMBV.

SUBMITTER: Yao JY

PROVIDER: S-EPMC8931665 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature

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Oral Vaccination With Recombinant Pichia pastoris Expressing Iridovirus Major Capsid Protein Elicits Protective Immunity in Largemouth Bass (Micropterus salmoides).

Yao Jia-Yun JY Zhang Cheng-Sai CS Yuan Xue-Mei XM Huang Lei L Hu Da-Yan DY Yu Zhe Z Yin Wen-Lin WL Lin Ling-Yun LY Pan Xiao-Yi XY Yang Gui-Lian GL Wang Chun-Feng CF Shen Jin-Yu JY Zhang Hai-Qi HQ

Frontiers in immunology 20220304

Largemouth bass iridovirus (LMBV) can cause high mortality and lead to heavy economic loss in the cultivation of largemouth bass, but there was no effective treatment. Here, the present study constructed a recombinant Pichia pastoris expressing LMBV major capsid protein (MCPD). The recombinant GS115-pW317-MCPD was then used to immunize largemouth bass via oral administration, and mucosal immune response mediated by immunoglobulins (Igs) was measured after oral immuniz ...[more]

PMID: 35309312

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Project description:BackgroundOocyte maturation in fish involves numerous cell signaling cascades that are activated or inhibited during specific stages of oocyte development. The objectives of this study were to characterize molecular pathways and temporal gene expression patterns throughout a complete breeding cycle in wild female largemouth bass to improve understanding of the molecular sequence of events underlying oocyte maturation.MethodsTranscriptomic analysis was performed on eight morphologically diverse stages of the ovary, including primary and secondary stages of oocyte growth, ovulation, and atresia. Ovary histology, plasma vitellogenin, 17β-estradiol, and testosterone were also measured to correlate with gene networks.ResultsGlobal expression patterns revealed dramatic differences across ovarian development, with 552 and 2070 genes being differentially expressed during both ovulation and atresia respectively. Gene set enrichment analysis (GSEA) revealed that early primary stages of oocyte growth involved increases in expression of genes involved in pathways of B-cell and T-cell receptor-mediated signaling cascades and fibronectin regulation. These pathways as well as pathways that included adrenergic receptor signaling, sphingolipid metabolism and natural killer cell activation were down-regulated at ovulation. At atresia, down-regulated pathways included gap junction and actin cytoskeleton regulation, gonadotrope and mast cell activation, and vasopressin receptor signaling and up-regulated pathways included oxidative phosphorylation and reactive oxygen species metabolism. Expression targets for luteinizing hormone signaling were low during vitellogenesis but increased 150% at ovulation. Other networks found to play a significant role in oocyte maturation included those with genes regulated by members of the TGF-beta superfamily (activins, inhibins, bone morphogenic protein 7 and growth differentiation factor 9), neuregulin 1, retinoid X receptor, and nerve growth factor family.ConclusionsThis study offers novel insight into the gene networks underlying vitellogenesis, ovulation and atresia and generates new hypotheses about the cellular pathways regulating oocyte maturation.

Project description:In order to improve the glucose utilization capacity of largemouth bass (Micropterus salmoides), responses to glucose overload between two strains (Y: breeding strain; W: wild strain) were compared at 0, 6, 12, and 24 h after glucose injection (1.67 g/kg). The data revealed that plasma glucose in the Y strain (<12 h) recovered faster than in the W strain (12 h), with the Y strain secreted more insulin within 6 h post-injection. Triglyceride (TG) and low-density lipoprotein-cholesterol (VLDL-CH) content in the Y strain increased, peaking at 12 h, then decreased, whereas the W strain's TG content was not affected and VLDL-CH content decreased. The hepatic and muscular fatty acid synthetase, liver x receptor-1, and sterol regulatory element-binding protein expressions were consistent with the TG content change. Both strains' liver and muscle glycogen contents exhibited similar trends to that of the glycogen synthase gene-increasing, then declining, and peaking at 6 and 12 h. The expression levels of hepatic and muscular phosphofructokinase and pyruvate kinase in the Y strain increased, peaking at 12 h. In the W strain, they were suppressed and reached the minimum at 24 h. The mRNA levels of hepatic and muscular phosphoenolpyruvate carboxykinase and glucose-6-phosphatase were enhanced and peaked at 24 h in both strains, hepatic isocitrate dehydrogenase-1, and α-ketoglutarate dehydrogenase complex expression increased after declining, peaking at 12 and 24 h. Two genes in the W strain's muscles showed a similar trend. Both strains' transcriptome results identified seven common functional genes for resistance to hyperglycemia that were involved in the circadian rhythm pathway, which is a suggested key pathway for coping with hyperglycemia. Furthermore, 48 differential genes were identified between the two strains, and these genes were enriched in the TGF-beta and cell cycle signaling pathways, indicating that these pathways may be key factors affecting the differential responses to glucose overload. We conducted a comprehensive comparison of glucose overload molecular responses between two strains of M. salmoides, and the results can provide a promising strategy to improve the glucose utilization capacity of M. salmoides based on advantageous pre-existing traits.

Dataset Information

Oral Vaccination With Recombinant Pichia pastoris Expressing Iridovirus Major Capsid Protein Elicits Protective Immunity in Largemouth Bass (Micropterus salmoides).

Publications

Oral Vaccination With Recombinant <i>Pichia pastoris</i> Expressing Iridovirus Major Capsid Protein Elicits Protective Immunity in Largemouth Bass (<i>Micropterus salmoides</i>).

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