thymosin and Fish-Diseases

Thymosin β4 is a multifunctional protein in vertebrates that participates in physiological processes, such as wound healing, immune response, cell proliferation and migration. We assessed the multifarious roles of this small peptide in Pinctada fucata, an oyster commonly used in pearl culture in China. Our results showed that when P. fucata was challenged by bacterial pathogens or LPS, the relative expression level of Pfthymosin β4 mRNA was significantly up-regulated, suggesting its involvement in immune response of the animal. Recombinant Pfthymosin β4 (rPfthymosin β4) was produced and showed in vitro different antibacterial activities against different pathogenic bacteria; the inhibitory effect of rPfthymosin β4 on bacterial growth was relatively stronger in the broth culture than agar culture. The overexpression of Pfthymosin β4 in Escherichia coli BL21(DE3) cells could improve their resistance to Cu

Topics: Animals; Aquaculture; Fish Diseases; Lipopolysaccharides; Pinctada; Streptococcal Infections; Streptococcus agalactiae; Thymosin; Vibrio; Vibrio alginolyticus; Vibrio Infections

Thymosin hormones, which were shown to be involved in immune system development and differentiation in previous studies, have antimicrobial functions in different animals. Zebrafish are a useful model for immunology research. Although thymosin has been reported to be involved in the embryonic development of zebrafish, it is necessary to uncover the antimicrobial function of thymosin in zebrafish. In this study, we expressed thymosin β (Tβ) in zebrafish in vitro and studied its antimicrobial function. The Tβ protein consists of 45 amino acids and is conserved among its family members, especially the actin-binding motif (LKKTET). Tβ was expressed in all tested tissues and was highly expressed in the brain, liver and hindgut. After Aeromonas hydrophila challenge, the Tβ transcript level increased in the skin, liver, kidney, spleen, thymus, foregut, gills and midgut. Purified recombinant thymosin β (rTβ) protein was used to study the antimicrobial mechanism. rTβ could inhibit the growth of Staphylococcus aureus, Aeromonas hydrophila, Vibrio anguillarum, Pseudomonas aeruginosa and Klebsiella pneumoniae. rTβ also binds to and agglutinates certain bacteria. Further study showed that rTβ could combine with the polysaccharides from gram-negative and gram-positive bacterial walls. All results suggested that the Tβ of zebrafish plays a significant role in innate antibacterial immune responses.

Topics: Aeromonas hydrophila; Animals; Fish Diseases; Fish Proteins; Gram-Negative Bacterial Infections; Immunity, Innate; Thymosin; Zebrafish

Thymosin beta belongs to the thymosin family, which consists of a series of highly conserved peptides involved in various biological processes. In teleosts, understanding of the immunological functions of thymosin beta is limited, particularly in vivo, which is essentially unknown. In the current study, we cloned and identified thymosin beta 4 from the teleost fish Golden pompano (Trachinotus ovatus), which we have named TroTβ4. We investigated the expression patterns and functions of TroTβ4 in both in vivo and in vitro assays. TroTβ4 is composed of 44 amino acids and shares high sequence identities with known thymosin β4 species in other teleosts, which contains a highly conserved actin-binding motif (LKKTET). The expression of TroTβ4 was most abundant in immune organs, and was significantly up-regulated in response to infection bacterial with one of a number of bacteria (including Edwardsiella tarda, Vibrio harveyi, and Streptococcus agalactiae). Purified recombinant TroTβ4 (rTroTβ4) inhibited the growth of bacteria, as measured using an automatic growth curve analyzer, indicating that TroTβ4 has antimicrobial functions. When administered in vivo, overexpression of TroTβ4 in T. ovatus, bacterial colonization of tissues was significantly reduced. In contrast, when a DNA vector-based siRNA technology was used to knock down TroTβ4 expression, bacterial dissemination and colonization of tissues increased significantly. Taken together, these results provide the first in vivo evidence to indicate that teleost thymosin beta 4 plays a significant role in innate antibacterial immune responses in addition to in vitro bacteriostatic activity. This provides valuable information regarding the biological functions of teleost thymosin beta.

Topics: Amino Acid Sequence; Animals; Base Sequence; Edwardsiella tarda; Enterobacteriaceae Infections; Fish Diseases; Fish Proteins; Immunity, Innate; Perciformes; Phylogeny; Sequence Alignment; Streptococcal Infections; Streptococcus agalactiae; Thymosin; Vibrio; Vibrio Infections

Prothymosin alpha (ProTα) is a small protein that in mammals is known to participate in diverse biological processes including immunomodulation. In teleost, the immunological function of ProTα is unknown. In the current study, we investigated the expression and function of the ProTα (named CsProTα) from the teleost fish tongue sole (Cynoglossus semilaevis). We found that CsProTα expression was abundant in immune relevant tissues and upregulated by megalocytivirus infection. Immunoblot detected secretion of CsProTα by peripheral blood leukocytes. Recombinant CsProTα (rCsProTα) as well as the C-terminal 11-residue (Ct11) were able to bind head kidney monocytes (HKM) and induce immune gene expression; however, the induction patterns caused by rCsProTα and Ct11 differed considerably. When introduced in vivo, rCsProTα and Ct11 significantly reduced megalocytivirus infection in fish tissues, whereas rCsProTα antibody significantly promoted viral replication. Blocking of Myd88 activity abolished the virus-inhibitory effect of rCsProTα but not Ct11. Taken together, these results demonstrate for the first time that both the intact protein and the C-terminal segment of a teleost ProTα can act like cytokines and induce antiviral immunity via, however, distinct signaling pathways that differ in the requirement of Myd88.

Topics: Amino Acid Sequence; Animals; Base Sequence; Cytokines; Fish Diseases; Flatfishes; Iridoviridae; Molecular Sequence Data; Myeloid Differentiation Factor 88; Protein Precursors; Recombinant Proteins; Sequence Alignment; Sequence Analysis, DNA; Signal Transduction; Thymosin