The effect of kynurenic acid on the synthesis of selected cytokines by murine splenocytes - in vitro and ex vivo studies

Abstract

Kynurenic acid (KYNA), a secondary product of the kynurenine pathway of tryptophan degradation, known mainly as an endogenous neuroprotectant, shows also immunotropic properties. Some quantities of KYNA are present in food and are effectively absorbed in the gastrointestinal tract. Since the spleen is an important target of dietary immunomodulators, the aim of the study was to determine the effect of exogenous KYNA on murine splenocytes. Splenocytes isolated from adult BALB/c mice were used in the study. Firstly, the effect of increasing KYNA concentrations (0-5 mM) on the viability, and proliferative and cytokine response (interleukin 1β [IL-1β], IL-6, IL-10, tumor necrosis factor α [TNF-α]) of murine splenocytes under in vitro conditions was determined. Then, proliferative and cytokine responses were determined in cells derived from animals receiving kynurenic acid in drinking water at concentrations of 2.5, 25, or 250 mg/l for 7-14 days. Cytokine levels were measured using commercial immunoassay (ELISA) kits, and cell viability and proliferation was determined with MTT reduction assay. Exogenous KYNA was characterised by a low level of cytotoxicity towards murine splenocytes, and was well tolerated by the animals receiving it in drinking water. As expected, it exhibited anti-inflammatory action towards the activated splenocytes, under both in vitro and ex vivo conditions. Surprisingly, however, KYNA itself influenced the activity of resting, non-stimulated cells, exerting an immunostimulant effect in vitro, and an immunosuppressive effect under ex vivo conditions. The obtained results indicate not only anti-inflammatory, but also more complex, immunomodulating properties of KYNA, which require more detailed investigation.

Keywords: cytokine response; immunomodulating properties; kynurenic acid; murine splenocytes.

Fig. 1

The in vitro effect of KYNA on the viability (A) and proliferative response (B) of murine splenocytes (n = 8)

Fig. 2

The in vitro effect of KYNA on the cytokine production by unstimulated (A) and LPS-stimulated splenocytes (B) (n = 8)

Fig. 3

Relative changes in cytokine production by murine splenocytes after oral administration of kynurenic acid (n = 6). A, B) Unstimulated cells, administration time 7 and 14 days, respectively; C, D) cells stimulated with bacterial LPS, administration time 7 and 14 days, respectively