Abstract
Tumor necrosis factor (TNF) plays an important role in an inflammatory cytokine storm. Over-secretion of TNF by the host in response to infection aggravates the disease. TNF expression level is positively correlated with the mortality caused by some bacterial infections. Therefore, using TNF antibody may alleviate the inflammation to resist bacterial infections. The function of fish TNF-b antibody in bacterial infection is still unclear. In this study, infection models of Vibrio vulnificus FJ03-X2 strain with high pathogenicity and strong virulence were established in zebrafish (Danio rerio) fibroblast cell line (ZF4 cells) and zebrafish. Zebrafish tnfb (Zetnf-b) gene was cloned and expressed by Escherichia coli BL21 (DE3), and Zetnf-b polyclonal antibody was prepared. Pre-injection of Zetnf-b polyclonal antibody and AG-126 before infecting with V. vulnificus could increase the survival rate of zebrafish by 36.6 and 46.7%, respectively. Pre-injection of Zetnf-b polyclonal antibody could effectively decrease the mortality of zebrafish infected by V. vulnificus. Thus, TNF polyclonal antibody therapy could be considered as an effective strategy to control V. vulnificus in fish.
Highlights
Fish tumor necrosis factor (TNF) family members are cytokines with multiple effects, which play vital roles in cell growth, differentiation, proliferation and immunity, apoptosis, macrophage activity and autoimmune diseases [1]
This study aimed to investigate the function of zebrafish TNF-b polyclonal antibody in fighting against V. vulnificus infection
The cytotoxic effect of V. vulnificus determined by evaluation lactate dehydrogenase (LDH) activity released from ZF4 cells is shown in Table 2 and Figure 1B
Summary
Fish tumor necrosis factor (TNF) family members are cytokines with multiple effects, which play vital roles in cell growth, differentiation, proliferation and immunity, apoptosis, macrophage activity and autoimmune diseases [1]. Most of the TNF family members in fish are similar to mammals, but some homologs are unique in fish. Multiple isoforms of TNF subtypes can be found in several fish species due to a genome duplication event in bony fish [2, 3]. TNF-alpha (TNF-α) is one of the important members of the TNF family, which has been identified in a variety of fish species, such as zebrafish (Danio rerio) [4], large yellow croaker (Larimichthys crocea) [5], Japanese flounder (Paralichthys olivaceus) [6] and grouper (Epinephelus coioides) [7]. Two copies of TNF-α have been cloned in orange-spotted grouper (E. coioides)
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