TAT-mediated oral subunit vaccine against white spot syndrome virus in crayfish.
White spot syndrome virus is a highly pathogenic virus that infects crayfish and other crustaceans. VP28 is one of its major envelope proteins, and plays a crucial role in viral infection. Cell-penetrating peptides are short peptides that facilitate cellular uptake of various molecular cargoes, and one well known example is TAT peptide from HIV-1 TAT protein. In this study, recombinant plasmids were constructed and transformed into Escherichia coli strain BL21 (DE3) to express TAT-VP28, VP28, TAT-VP28-EGFP and VP28-EGFP fusion proteins. Enzyme-linked immunosorbent assay (ELISA) and flow cytometry methods were used to confirm that TAT fusion proteins can translocate from the intestine to the hemolymph of the crayfish Cambarus clarkii. After immunization, activities of phenoloxidase and superoxide dismutase were analyzed, and it was found that rTAT-VP28 produced the most pronounced increase in both C. clarkii were vaccinated by oral administration of rTAT-VP28 and rVP28 for 7 and 14 days, and rTAT-VP28 resulted in the highest relative percent survival (RPS) (63.3% at 7 days, and 67.8% at 14 days), compared with rVP28 (44.4% at 7 days, and 53.6% at 14 days) following challenge with WSSV after the last day of feeding. This study reports the use of TAT-derived peptide as an oral delivery method of a subunit vaccine against WSSV in C. clarkii.
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, China. zhang000yi@126.com
Yi
Y
Zhang
Jian-Fang
JF
Ning
Xing-qin
XQ
Qu
Xiao-Lin
XL
Meng
Jin-Ping
JP
Xu
D000818:Animals; D003400:Astacoidea / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005434:Flow Cytometry; D015870:Gene Expression; D006458:Hemolymph / Q000737:chemistry; D007422:Intestines / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012758:Shellfish; D016019:Survival Analysis; D022223:Vaccines, Subunit / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology; D014765:Viral Vaccines / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology*; D046848:White spot syndrome virus 1 / Q000276:immunology*; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000276:immunology
D011993:Recombinant Fusion Proteins; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; D054322:tat Gene Products, Human Immunodeficiency Virus