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ABSTRACT
Repulsive guidance molecule a (RGMa) is an axonal guidance molecule that has recently found to exert function in immune system. This study evaluated the function of RGMa in modulation of dendritic cells (DCs) function stimulated with Achyranthes bidentata polysaccharide (ABP) and lipopolysaccharide (LPS) using a RGMa-neutralizing antibody. Compared with the Control-IgG/ABP and Control-IgG/LPS groups, DCs in the Anti-RGMa/ABP and Anti-RGMa/LPS groups 1) showed small, round cells with a few cell processes and organelles, and many pinocytotic vesicles; 2) had decreased MHC II, CD86, CD80, and CD40 expression; 3) displayed the decreased IL-12p70, IL-1β and TNF-α levels and increased IL-10 secretion; 4) had a high percentage of FITC-dextran uptake; and 5) displayed a reduced ability to drive T cell proliferation and reinforced T cell polarization toward a Th2 cytokine pattern. We conclude that DCs treated with RGMa-neutralizing antibodies present with tolerogenic and immunoregulatory characteristics, which provides new insights into further understanding of the function of RGMa.
Abbreviations
| BMDC | = | bone-marrow derived dendritic cell |
| CNS | = | central nervous system |
| DCs | = | dendritic cells |
| EAE | = | experimental autoimmune encephalomyelitis |
| ELISA | = | enzyme-linked immunosorbent assay |
| FACS | = | fluorescence activated cell sorting |
| GM-CSF | = | granulocyte macrophage-colony stimulating factor |
| HRP | = | horseradish peroxidase |
| IRAKs | = | interleukin receptor-associated kinases |
| LPS | = | lipopolysaccharide |
| MACS | = | magnetic activated cell sorting |
| MAPK | = | mitogen-activated protein kinase |
| MD2 | = | myeloid differentiation protein 2 |
| MHC | = | major histocompatibility complex |
| MLR | = | Mixed lymphocyte reaction |
| MOG | = | myelin oligodendrocyte glycoprotein |
| MS | = | multiple sclerosis |
| MyD88 | = | myeloid differentiation factor 88 |
| NF-κB | = | nuclear factor kappa-B |
| OD | = | optical density |
| PBS | = | phosphate balanced solution |
| PI | = | propidium iodine; MTS, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt |
| PVDF | = | polyvinylidene difluoride |
| RBCs | = | red blood cells |
| RGMa | = | repulsive guidance molecule a |
| RIPA | = | radio-immunoprecipitation assay |
| SDS-PAGE | = | sodium dodecyl sulfate-polyacrylamide gel electrophoresis |
| SEM | = | scanning electron microscopy |
| SOCS1 | = | suppressor of cytokine signaling 1 |
| TEM | = | transmission electron microscopy |
| TIR | = | interleukin 1 receptor |
| TIRAP | = | TIR domain-containing adaptor protein |
| TLR4 | = | Toll-like receptor 4 |
| TRAF6 | = | TNF receptor-associated factor 6 |
| TRAM | = | TRIF-related adaptor molecule |
| TRIF | = | TIR domain-containing adapter-inducing IFN β |
Disclosure of potential conflicts of interest
No potential conflicts of interest were disclosed.
Acknowledgments
This work was supported financially by Liaoning Province Science and Technology Project-Animal Scientific Research and Clinical Application for Major Disease of Liaoning Province (2012225021) and Technology Projects of Liaoning Province (2009225010-2) to Juan Feng.