thymosin and Enterobacteriaceae-Infections

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

Animal models for opportunistic infections were developed by using mice immunosuppressed by 5-FU. These mice were susceptible to various microorganisms, while normal mice had greater tolerance to such microbial infections. In these models, thymosin alpha 1 was found to protect mice against lethal infections with Candida albicans, Listeria monocytogenes, Pseudomonas aeruginosa, and Serratia marcescens when it was administered during 5-FU treatment prior to the infections. Thymosin alpha 1 was effective in some infections at 0.4-400 micrograms/kg/day IP, about 1/100 of the dose required for thymosin fraction 5. Activity was also demonstrated against L-monocytogenes and Ps. aeruginosa by counting the viable bacteria in the liver after infection. The protective activity against Candida, elimination of which macrophages were essential, was abrogated by anti-thymocyte serum and/or carrageenan, indicating that thymosin alpha 1 serves to maintain the functions of macrophages by reducing the damage to T cells by 5-FU. On the other hand, the activity against Pseudomonas infection was not affected by anti-thymocyte serum or carrageenan. It is probable that thymosin alpha 1 also exerts its effect on neutrophils without participation of T cells and macrophages.

Topics: Animals; Candida albicans; Candidiasis; Drug Administration Schedule; Enterobacteriaceae Infections; Fluorouracil; Listeria monocytogenes; Listeriosis; Mice; Mice, Inbred Strains; Pseudomonas Infections; Serratia marcescens; Thymosin; Thymus Hormones