Interaction of bone marrow stromal cells with Kupffer cells induces immune tolerance

Abstract

Objective To investigate the phenomenon of bone marrow stromal cells (BMSCs) regulating the function of Kupffer cells (KCs) under direct contact co-culture in vitro.Methods In co-culture experiments,pure rat BMSCs were directly added to the KCs plates with the ratio of 1∶ 1.KCs were separately cultured in a well as control group.After stimulation with lipopolysaccharide (LPS,1 mg/L) for 24 h,the gene and protein expression of interleukin (IL)-10 was detected by using real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting,respectively.The abundance of IL-10,prostaglandin E2 (PGE2) and IL-6 was detected by using enzyme linked immunosorbent assay (ELISA) kits.Flow cytometry was used to detect the expression of major histocompatibility complex (MHC) Ⅱ,CD40,CD80 and CD86.Results As compared with control group,the gene and protein expression of IL-10 in co-culture system was significantly increased (P ＜ 0.05).After co-culture with BMSCs,the levels of IL-10 [KCs:(1000.12±49.06) ng/L; KCs + BMSCs:(2307.44 ±308.97) ng/L] and PGE2[KCs:(244.48 ±28.15) ng/L; KCs + BMSCs:(424.48 ± 47.86) ng/L] were significantly increased (P ＜ 0.01),while the abundance of IL-6 [KCs:(283.18 ± 20.50) ng/L; KCs + BMSCs:(78.14 ± 10.58) ng/L] was significantly down-regulated (P ＜ 0.01).In co-culture system,the expression of MHC Ⅱ,CD40,CD80 and CD86 was lower than in control group.Conclusion BMSCs may reprogram the expression patterns of KCs,regulate the function of KCs,and then induce immune tolerance. Key words: Bone marrow stromal cells; Kupffer cells; Co-culture; Reprogram