The immunomodulatory effects of human mesenchymal stem cells on peripheral blood mononuclear cells in ALS patients

In a previous study, we reported that intrathecal injection of mesenchymal stem cells (MSCs) slowed disease progression in G93A mutant superoxide dismutase1 transgenic mice. In this study, we found that intrathecal MSC administration vastly increased the infiltration of peripheral immune cells into the spinal cord of Amyotrophic lateral sclerosis (ALS) mice (G93A mutant superoxide dismutase1 transgenic). Thus, we investigated the immunomodulatory effect of MSCs on peripheral blood mononuclear cells (PBMCs) in ALS patients, focusing on regulatory T lymphocytes (T-reg; CD4(+)/CD25(high)/FoxP3(+)) and the mRNA expression of several cytokines (IFN-, TNF-, IL-17, IL-4, IL-10, IL-13, and TGF-). Peripheral blood samples were obtained from nine healthy controls (HC) and sixteen patients who were diagnosed with definite or probable ALS. Isolated PBMCs from the blood samples of all subjects were co-cultured with MSCs for 24 or 72h. Based on a fluorescence-activated cell sorting analysis, we found that co-culture with MSCs increased the T-reg/total T-lymphocyte ratio in the PBMCs from both groups according to the co-culture duration. Co-culture of PBMCs with MSCs for 24h led to elevated mRNA levels of IFN- and IL-10 in the PBMCs from both groups. However, after co-culturing for 72h, although the IFN- mRNA level had returned to the basal level in co-cultured HC PBMCs, the IFN- mRNA level in co-cultured ALS PBMCs remained elevated. Additionally, the levels of IL-4 and TGF- were markedly elevated, along with Gata3 mRNA, a Th2 transcription factor mRNA, in both HC and ALS PBMCs co-cultured for 72h. The elevated expression of these cytokines in the co-culture supernatant was confirmed via ELISA. Furthermore, we found that the increased mRNA level of indoleamine 2,3-dioxygenase (IDO) in the co-cultured MSCs was correlated with the increase in T-reg induction. These findings of T-reg induction and increased anti-inflammatory cytokine expression in co-cultured ALS PBMCs provide indirect evidence that MSCs may play a role in the immunomodulation of inflammatory responses when MSC therapy is targeted to ALS patients. We propose the following mechanism for the effect of mesenchymal stem cells (MSCs) administered intrathecally in amyotrophic lateral sclerosis (ALS): MSCs increase infiltration of peripheral immune cells into CNS and skew the infiltrated immune cells toward regulatory T lymphocytes (T-reg) and Th2 lymphocytes. T-reg and Th2 secret anti-inflammatory cytokines such as IL-4, IL-10, and TGF-. A series of immunomodulatory mechanism provides a new strategy for ALS treatment.This study was supported by grants from the Korea Healthcare Technology R&D Project of the Ministry for Health & Welfare Affairs of the Republic of Korea (A101712 and A120182). The authors have no conflicts of interest to disclose