Amelioration of rat collagen-induced arthritis through CD4+ T cells apoptosis and synovial interleukin-17 reduction by indoleamine 2,3-dioxygenase gene therapy.

Indoleamine 2,3-dioxygenase (IDO) has been known as an emerging therapeutic target in autoimmunity-related arthritis. The treatment responses of adenoviral vectors encoding IDO (AdIDO) gene therapy in rat collagen-induced arthritis (CIA) were examined in this study. The therapeutic effects on ankle circumference, articular index, and radiographic and histological scores were evaluated in AdIDO-injected ankle joints. We further determined CD4+ T-cell numbers and their apoptotic status, CD68(+) macrophage numbers, kynurenine (a downstream tryptophan metabolite) concentrations, interleukin-17 (IL-17) levels, and retinoic acid-related orphan receptor γt (RORγt) expression in synovial tissues of CIA rats receiving AdIDO treatment. Reduction of ankle circumference, articular index, and radiographic and histological scores were noted in AdIDO-treated ankles, as compared with those receiving injection of control vectors. Furthermore, IDO gene transfer led to decreased infiltrating CD4+ T cells with enhanced apoptosis, reduced CD68+ macrophage numbers, increased kynurenine levels, lower IL-17 concentrations, and decreased RORγt expression within the ankle joints. In addition, such a therapy diminished type II collagen-specific IL-17 production and RORγt expression in CD4+ T cells from draining lymph nodes of CIA rats. Our results demonstrate for the first time that intra-articular delivery of IDO gene ameliorated ankle arthritis of CIA rats by induction of CD4+ T-cell apoptosis and reduction of synovial IL-17 production through the supplement of kynurenine. Taken together, these findings implicate the novel strategy of using IDO gene as a therapeutic approach in treating patients with rheumatoid arthritis.

Effect of Kidney-replenishing herb on the indoleamine 2,3-dioxygenase of human syncytiotrophoblasts cultured in vitro and the balance of helper T-cell cytokines.

BACKGROUND: There is complicated pathogeny involved in spontaneous abortion. At present, the focus of study is on the interface between mother and fetus, the trophoblasts. Indoleamine 2,3-dioxygenase (IDO) is the first and regulatory enzyme in the major route of l-tryptophan catabolism, which induces immunosuppression of T lymphocytes. In the present study we investigated the effect of Kidney-replenishing herb on the expression and activity of IDO in human syncytiotrophoblasts cultured in vitro and the balance of helper T cell (Th) cytokines. METHODS: Syncytiotrophoblasts were cultured in vitro for 24, 48 or 72 h, with either control serum or serum made from Kidney-replenishing herb, without or with different concentrations of the IDO inhibitor 1-methyltryptophan (1-MT). Reverse transcription-polymerase chain reaction was applied to analyze the IDO mRNA transcription of syncytiotrophoblasts and Western blotting was applied to determine the expression of IDO protein in syncytiotrophoblasts. The concentration of interleukin-10 and interferon-gamma in co-culture medium of syncytiotrophoblasts and decidual T lymphocytes was determined by enzyme-linked immunosorbent assay. High-performance liquid chromatography was used to determine the concentration of kynurenine (Kyn) and tryptophan (Tyr) in the co-culture medium, and the ratio of Kyn/Try was used to assess IDO activity. RESULTS: IDO mRNA and protein were detected in human syncytiotrophoblasts cultured in vitro. The IDO inhibitor 1-MT caused the balance of Th cytokines to depart from type 2; when IDO activity was inhibited, Kidney-replenishing herb improved the expression of IDO mRNA and protein, promoted IDO activity and caused the balance of Th cytokines depart from type 1. CONCLUSION: Kidney-replenishing herb improves the expression of IDO mRNA and protein, promotes IDO activity to an appropriate value, resumes the balance of Th cytokines and regulates maternofetal tolerance.

Simultaneous measurements of tryptophan and its metabolites, kynurenine and serotonin, in the superficial layers of the spinal dorsal horn. A study in normal and arthritic rats.

It has been demonstrated that 5-hydroxytryptamine (5-HT) is not the only neuroactive metabolite of tryptophan (TRP) in the CNS. The presence of kynurenine (KYN) and its metabolites has been reported in the brain of several mammalian species and the neuroactive properties of these compounds are now well established. In the present study, we report the identification of KYN in the superficial layers of the rat spinal dorsal horn. KYN was measured simultaneously with TRP. 5-hydroxytryptophan, 5-HT, 5-hydroxyindoleacetic acid and 5-HT-O-sulfate by means of liquid chromatography with coulometric electrode array detection. The results observed in the normal rat and in an animal model of persistent pain, the arthritic rat, are discussed in view of the hypothesis relating to the involvement of the bulbospinal serotonergic system in pain mechanisms and of the possible participation of KYN and its metabolites in these mechanisms.