J Korean Med Sci.  2017 Sep;32(9):1502-1507. 10.3346/jkms.2017.32.9.1502.

The Anti-Inflammatory Effects of Oral-Formulated Tacrolimus in Mice with Experimental Autoimmune Encephalomyelitis

• Kim MJ
• Sung JJ
• Kim SH
• Kim JM
• Jeon GS
• Mun SK
• Ahn SW

• KMID: 2385958
• DOI: http://doi.org/10.3346/jkms.2017.32.9.1502

Abstract

Multiple sclerosis (MS) is a T-lymphocyte-mediated autoimmune disease that is characterized by inflammation in the central nervous system (CNS). Although many disease-modifying therapies (DMTs) are presumed effective in patients with MS, studies on the efficacy and safety of DMTs for preventing MS relapse are limited. Therefore, we tested the immunosuppressive anti-inflammatory effects of oral-formulated tacrolimus (FK506) on MS in a mouse model of experimental autoimmune encephalomyelitis (EAE). The mice were randomly divided into 3 experimental groups: an untreated EAE group, a low-dose tacrolimus-treated EAE group, and a high-dose tacrolimus-treated EAE group. After autoimmunization of the EAE mice with myelin oligodendrocyte glycoprotein, symptom severity scores, immunohistochemistry of the myelination of the spinal cord, and western blotting were used to evaluate the EAE mice. After the autoimmunization, the symptom scores of each EAE group significantly differed at times. The group treated with the larger tacrolimus dose had the lowest symptom scores. The tacrolimus-treated EAE groups exhibited less demyelination and inflammation and weak immunoreactivity for all of the immunization biomarkers. Our results revealed that oral-formulated tacrolimus inhibited the autoimmunization in MS pathogenesis by inactivating inflammatory cells.

Keyword

Multiple Sclerosis; Neuromyelitis Optica; Experimental Autoimmune Encephalomyelitis; EAE; Tacrolimus; FK506

MeSH Terms

Animals
Autoimmune Diseases
Biomarkers
Blotting, Western
Central Nervous System
Demyelinating Diseases
Encephalomyelitis, Autoimmune, Experimental*
Humans
Immunization
Immunohistochemistry
Inflammation
Mice*
Multiple Sclerosis
Myelin Sheath
Myelin-Oligodendrocyte Glycoprotein
Neuromyelitis Optica
Recurrence
Spinal Cord
Tacrolimus*
Biomarkers
Myelin-Oligodendrocyte Glycoprotein
Tacrolimus

Figure

• 

  Fig. 1 Behavioral tests of EAE mice. After autoimmunization with MOG35–55, the symptom scores of each EAE group gradually increased over time, and significant differences were found among the 3 groups (P < 0.05) beginning on the 10th day. The 10-mg/kg tacrolimus-treated EAE mice exhibited lower EAE scores, which suggested that they had less clinical symptoms, compared with the untreated EAE mice and 5-mg/kg tacrolimus-treated EAE mice.EAE = experimental autoimmune encephalomyelitis, MOG = myelin oligodendrocyte glycoprotein.

• 

  Fig. 2 Histopathology of EAE mice. The spinal cord specimens of the untreated EAE mice exhibited marked demyelination (A) and inflammatory cell infiltration in the perivascular area (D). However, the tacrolimus-treated EAE mice exhibited preserved myelination and decreased inflammation compared with the untreated group. Additionally, the 10-mg/kg tacrolimus-treated EAE mice exhibited more myelination (C) and decreased inflammation (F) compared with the 5-mg/kg tacrolimus-treated EAE mice (B, E).EAE = experimental autoimmune encephalomyelitis.

• 

  Fig. 3 Western blots of EAE mice. Fourteen days after the immunization, specimens from the untreated EAE group and tacrolimus-treated groups were investigated with western blots. The blots of the samples from the tacrolimus-treated EAE mice contained bands with strong immunoreactivity to MOG and weak immunoreactivity to CD4 compared with those in the untreated mice (A). In addition, the blots of the samples from the tacrolimus-treated EAE mice had weak immunoreactivity to Iba1 and GFAP compared with those in the untreated EAE mice (B).EAE = experimental autoimmune encephalomyelitis, MOG = myelin oligodendrocyte glycoprotein, Iba1 = ionized calcium binding adaptor 1, GFAP = glial fibrillary acidic protein.

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