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Expression analysis of tissue factor pathway inhibitors TFPI-1 and TFPI-2 in Paralichthys olivaceus and antibacterial
Guanghua Wang1, Bing Xie1, Yanli Su1
1Marine Science and Engineering College, Qingdao Agricultural University, Qingdao, 266109, China.
Abstract:
Tissue factor pathway inhibitors (TFPI), including TFPI-1 and TFPI-2, are Kunitz-type serine protease inhibitors that mainly inhibit the blood coagulation induced by tissue factors. Previous reports on teleost proved TFPI play important roles in innate immunity. In this study, two TFPI (PoTFPI-1 and PoTFPI-2) molecules from Japanese flounder (Paralichthys olivaceus) were analyzed and characterized for their expression patterns, antibacterial and anticancer activities of the C-terminal derived peptides. Quantitative real time RT-PCR analysis shows that constitutive PoTFPI-1 expression occurred, in increasing order, in the brain, muscle, spleen, gills, head kidney, blood, intestine, heart, and liver; PoTFPI-2 was expressed, in increasing order, in the brain, gills, head kidney, muscle, intestine, spleen, liver, heart, and blood. Under the stimulation of fish pathogens, both PoTFPI-1 and PoTFPI-2 expressions increased significantly in a manner that depended on the pathogens, tissue type, and infection stage. Furthermore, C-terminal peptides TP25 and TP26, derived from PoTFPI-1 and PoTFPI-2, respectively, were synthesized and proved to be active against Micrococcus luteus (for TP25 and TP26) and Staphylococcus aureus (for TP25) via retardation effects on bacterial nucleic acids. In addition, TP25 and TP26 also displayed significant inhibitory effects on human colon cancer cell line HT-29. These results reveal that both PoTFPI-1 and PoTFPI-2 play important roles in host innate immunity. The antibacterial activity and anticancer cells function of TP25 and TP26 will add new insights into the roles of teleost TFPI.
Insights
Japanese flounder tissue factor pathway inhibitors (TFPI-1 and TFPI-2) are crucial for innate immunity. Their derived peptides, TP25 and TP26, exhibit potent antibacterial and anticancer activities, offering new insights into teleost immunity.
Area of Science:
- Immunology
- Marine Biology
- Biochemistry
Background:
- Tissue factor pathway inhibitors (TFPIs) are Kunitz-type serine protease inhibitors involved in blood coagulation.
- Previous studies indicate TFPIs play significant roles in teleost innate immunity.
- Japanese flounder (Paralichthys olivaceus) TFPI-1 and TFPI-2 were investigated for their immune functions.
Purpose of the Study:
- To characterize the expression patterns of PoTFPI-1 and PoTFPI-2 in Japanese flounder.
- To evaluate the antibacterial and anticancer activities of C-terminal derived peptides from PoTFPI-1 and PoTFPI-2.
- To elucidate the role of these TFPIs in the host's innate immune response.
Main Methods:
- Quantitative real-time RT-PCR was used to analyze PoTFPI-1 and PoTFPI-2 expression levels in various tissues.
- Expression levels were monitored under stimulation with fish pathogens.
- C-terminal peptides (TP25 and TP26) were synthesized and their biological activities were assessed.
Main Results:
- Constitutive expression of PoTFPI-1 and PoTFPI-2 was observed in multiple Japanese flounder tissues, with varying levels.
- Pathogen stimulation significantly upregulated the expression of both PoTFPI-1 and PoTFPI-2.
- Synthesized peptides TP25 and TP26 demonstrated antibacterial activity against Micrococcus luteus and Staphylococcus aureus, and inhibited human colon cancer cells (HT-29).
Conclusions:
- PoTFPI-1 and PoTFPI-2 are integral components of the Japanese flounder's innate immune system.
- The C-terminal peptides TP25 and TP26 possess significant antibacterial and anticancer properties.
- These findings contribute novel insights into the multifaceted roles of teleost TFPIs in host defense.
