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VARIATIONS IN EXPRESSION OF ELEVEN IMMUNE RELATED GENES IN FLOUNDER(PARALICHTHYS OLIVACEUS) INJECTED WITH INACTIVATED VIBRIO ANGUILLARUM
Author(s): 
Pages: 1024-1033
Year: Issue:  5
Journal: Oceanologia et Limnologia Sinica

Keyword:  Paralichthys olivaceusinactivated Vibrio anguillarumimmersion immunizationimmune related genes;
Abstract: We prepared inactivated whole-cell vaccine of Vibrio anguillarum, and then vaccinated Paralichthys olivaceus in whole-cell vaccine immunization. Samples of spleen, head kidney and gill of P. olivaceus were collected at 0h, 4h, 8h, 12 h, 24 h, 48 h, 72 h, 96 h, 7d, and 14 d after immunization. Total RNA was extracted from spleen, head kidney and gill, and then the gene expression levels of TLR2(Toll-like receptor 2), TLR5 M, Myeloid differentiation factor 88(My D88), NF-κB(nuclear transcription factors), IL-6(interleukin-6), IFNγ(interferon γ), CXC(chemotactic factor), C3(complement factor), Hsp70(heat shock protein 70), cluster determinant 4(CD4) and nature killer enhancing factor(NKEF) were tested in q RT-PCR(quantitative reverse transcription-polymerase chain reaction). The results show that the expression quantities of immuno-related genes increased significantly after immunization except for TLR5 M and NKEF. The maximum expression of immune-related genes occurred in 24~72h after immunization, and was 2~12 times higher than those of control group. However, TLR5 M showed no significant difference; and the expression of NKEF showed down-regulation and reached peak at 24 h after immunization. The minimum expression was 0.49 times of control group. In spleen and head kidney, the maximum expression of NF-κB and CD4 was higher than that in gill. In spleen and gill, the maximum expression of IL-6, HSP70, and NKEF was higher than that in head kidney. The maximum expression of IFNγ, CXC, C3, and My D88 occurred in similar time and showed no significant difference in the three tissues. The fast change and high expression of IL-6 and HSP70 in all the three tissues may be applied for evaluation on fish soaking vaccine immunity. In addition to the main immune organs including spleen and head kidney, gill can be an important tissue for the detection. This method may be applied for future evaluation on the efficacy of vaccination and its molecular mechanism.
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