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Two bicistronic DNA vaccines against Vibrio anguillarum and the immune eff ects on flounder Paralichthys olivaceus

Journal of Oceanology and Limnology 2022年第2期40卷 786-804页

作者： Hanlin LI Jing XING Xiaoqian TANG Xiuzhen SHENG Heng CHI Wenbin ZHAN Laboratory of Pathology and Immunology of Aquatic Animals KLMMEOcean University of ChinaQingdao 266003China Laboratory for Marine Fisheries Science and Food Production Processes Pilot National Laboratory for Marine Science and Technology(Qingdao)Qingdao 266071China

Chemokines are cytokines that can promote the activation and migration of immune cells,and increase the recognition of antigen by antigen-presenting cells(APC).Previous studies showed that a DNA vaccine can induce humoral and cellular immune responses of flounder after *** explore the improvement of chemokines on the efficiency of Ompk vaccine,two bicistronic DNA candidate vaccines were constructed and the immune responses they induced in the flounder were investigated by reverse transcription polymerase chain reaction(RT-PCR),indirect immunofl uorescent assay(IFA),H&E staining,fl ow cytometry(FCM),and quantifi cational real-time polymerase chain reaction(qRT-PCR).pBudCE4.1 plasmid as an expression vector,bicistronic DNA vaccines encoding Ompk gene and CC-motif ligand 4 gene(p-Ompk-CCL4),or Ompk gene and CC-motif ligand 19 gene(p-Ompk-CCL19)were successfully *** results showed that two bicistronic DNA vaccines expressed Ompk protein of Vibrio anguillarum and CCL4/CCL19 proteins of fl ounder both in vitro and in *** immunization,a large number of leucocytes in muscle were recruited at the injection site in treatment *** constructed vaccines induced signifi cant increases in CD4-1^(+) and CD4-2^(+) T lymphocytes,and sIgM^(+) B lymphocytes in peripheral blood,spleen,and head *** percentage of T lymphocytes peaked on the 14^(th) post-vaccination day whereas that of B lymphocytes peaked in the 6^(th) post-vaccination ***,the expression profi les of 10 immune-related genes increased in muscles around the injection site,spleen,and head *** the challenge,p-Ompk-CCL4 and p-Ompk-CCL19 conferred a relative percentage survival(RPS)of 74.1%and 63.3%,respectively,higher than p-Ompk alone(40.8%).In conclusion,both CCL4 and CCL19 can improve the protection of p-Ompk via evoking local immune response and then humoral and cellular ***4 and CCL19 will be potential molecular adjuvants for use in DNA vaccines.

关键词： Vibrio anguillarum outer membrane protein k bicistronic DNA vaccines CC-motif ligand 4 CC-motif ligand 19 immune response

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