title: The effect of 1, 25(OH)2 D3 (calcitriol) alone and in combination with all-trans retinoic acid on ROR-γt, IL-17, TGF-β, and FOXP3 gene expression in experimental autoimmune encephalomyelitis.
abstract: OBJECTIVE
  It has been shown that calcitriol and all-trans retinoic acid (ATRA) have modulatory effects on the immune system. The present study investigates the synergistic effects of combination treatment of calcitriol and ATRA in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS).
  
  
  METHODS
  The mice were allocated to four preventive groups, each consisting of eight animals, ATRA (250 μg/mouse), calcitriol (100 ng/mouse), combination of ATRA and calcitriol (125  μg/mouse and 50 ng/mouse) and vehicle groups. EAE was induced by MOG35-55 peptide in female C57BL/6 mice. Treatments were initiated at day 1 before immunization and continued every other day throughout the study until the day 21 post-immunization. Splenocytes were isolated from EAE-induced mice and the expression of retinoic acid receptor-related orphan receptor gamma t (ROR-γt), Interleukin-17 (IL-17), transforming growth factor beta (TGF-β), and forkhead box P3 (FOXP3) genes was measured using real-time polymerase chain reaction.
  
  
  RESULTS
  The expression of FOXP3 and TGF-β genes in the splenocytes of combination-treated and calcitriol alone-treated mice was significantly increased compared to vehicle group (P < 0.05). The expression of ROR-γt and IL-17 genes in the splenocytes of ATRA, calcitriol and combination- treated mice was significantly reduced compared to those of vehicle- treated mice (P < 0.05). The relative expression level of ROR-γt was significantly (P < 0.05) lower in the combination group than in the mice treated by ATRA or calcitriol alone.
  
  
  CONCLUSIONS
  This study demonstrated that treatment with combination of calcitriol and ATRA can be considered as a new strategy for MS prevention and treatment.
authors.affiliation: a Department of Cellular and Molecular Nutrition , School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences , Tehran , Iran.
authors.forename: Karim
authors.initials: K
authors.lastname: Parastouei
authors.affiliation: b Department of Immunology, School of Public Health , Tehran University of Medical Sciences , Tehran , Iran.
authors.forename: Abbas
authors.initials: A
authors.lastname: Mirshafiey
authors.affiliation: c Department of Biostatistics , School of Public Health, Tehran University of Medical Sciences , Tehran , Iran.
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Eshraghian
authors.affiliation: d Department of Cellular and Molecular Nutrition , School of Nutritional Sciences and Dietetics, International Campus, Tehran University of Medical Sciences , Tehran , Iran.
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Shiri-Shahsavar
authors.affiliation: e Department of Virology , School of Public Health, Tehran University of Medical Sciences , Tehran , Iran.
authors.forename: Farid
authors.initials: F
authors.lastname: Solaymani-Mohammadi
authors.affiliation: a Department of Cellular and Molecular Nutrition , School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences , Tehran , Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Chahardoli
authors.affiliation: a Department of Cellular and Molecular Nutrition , School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences , Tehran , Iran.
authors.forename: Ehsan
authors.identifier: http://orcid.org/0000-0001-9258-1006
authors.initials: E
authors.lastname: Alvandi
authors.affiliation: a Department of Cellular and Molecular Nutrition , School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences , Tehran , Iran.|b Department of Immunology, School of Public Health , Tehran University of Medical Sciences , Tehran , Iran.
authors.forename: Ali Akbar
authors.initials: AA
authors.lastname: Saboor-Yaraghi
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000008:administration & dosage / Q000627:therapeutic use*; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000008:administration & dosage / Q000627:therapeutic use*; D002117:Calcitriol / Q000008:administration & dosage / Q000627:therapeutic use*; D004305:Dose-Response Relationship, Drug; D004359:Drug Therapy, Combination; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000188:drug therapy / Q000276:immunology / Q000378:metabolism / Q000517:prevention & control*; D005260:Female; D051858:Forkhead Transcription Factors / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D007274:Injections, Intraperitoneal; D020381:Interleukin-17 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D008810:Mice, Inbred C57BL; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011897:Random Allocation; D015203:Reproducibility of Results; D013154:Spleen / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D014212:Tretinoin / Q000008:administration & dosage / Q000627:therapeutic use*
chemical_list: D000893:Anti-Inflammatory Agents; D000894:Anti-Inflammatory Agents, Non-Steroidal; D051858:Forkhead Transcription Factors; C498833:Foxp3 protein, mouse; C098680:Il17a protein, mouse; D020381:Interleukin-17; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3; D012333:RNA, Messenger; C539901:Rorc protein, mouse; C508961:Tgfb1 protein, mouse; D053773:Transforming Growth Factor beta1; D014212:Tretinoin; D002117:Calcitriol
keywords: ATRA; Calcitriol; Experimental autoimmune encephalomyelitis; FOXP3; ROR-γt