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miR-141 and miR-200a, Revelation of New Possible Players in Modulation of Th17/Treg Differentiation and Pathogenesis of Multiple Sclerosis.

Naghavian R, Ghaedi K, Kiani-Esfahani A, Ganjalikhani-Hakemi M, Etemadifar M, Nasr-Esfahani MH - PLoS ONE (2015)

Bottom Line: Expression level of miR-141 and miR-200a were measured by RT-q PCR and compared to healthy control group (n=10).Furthermore, both miR-141 and miR-200a show up-regulation in relapsing phase of MS patients compared to remitting and control groups.Our data suggest that these miRNAs may probably inhibit negative regulators of Th17 cell differentiation, thus promoting its differentiation.

View Article: PubMed Central - PubMed

Affiliation: Division of Cellular and Molecular Biology, Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran; Department of Cellular Biotechnology at Cell Science research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.

ABSTRACT

Background: One of the main issues in pathogenesis of MS is Th17/Treg imbalance. There are growing interests in nominating miRNAs involved in Th17 cell differentiation, suggesting them as new therapeutic agents that may reduce progression of different autoimmune diseases specially MS.

Objectives: We assessed transcript levels of miR-141 and miR-200a in MS patients, during relapsing and remitting phases. We also investigated possible role of miR-141, miR-200a in inducing differentiation to Th17 cells.

Materials and methods: Forty RR-MS patient samples including relapsing (n=20) and remitting (n=20) phases were chosen. Expression level of miR-141 and miR-200a were measured by RT-q PCR and compared to healthy control group (n=10). In-silico analyses on miR-141 and miR-200a targetome showed involvement of both miRNAs in T helper cell differentiation pathways including TGF-β, mTOR and JAK/STAT.

Results: We observed that percentage of RORγt+ CD4+ T cells increase in relapsing phase while FOXP3+ CD4+ increase in remitting phase of MS patients. Furthermore, both miR-141 and miR-200a show up-regulation in relapsing phase of MS patients compared to remitting and control groups. Interestingly, expression level of target genes of miR-141 and miR-200a, which were assessed through in-silico methods, show down-regulation in relapsing phase of MS patients.

Conclusions: According to our results, miR-141 and miR-200a may be key miRNAs in progression of symptoms of MS through inducing differentiation of Th17 cells and inhibiting differentiation to Treg cells. Our data suggest that these miRNAs may probably inhibit negative regulators of Th17 cell differentiation, thus promoting its differentiation.

No MeSH data available.


Related in: MedlinePlus

Flow cytometry of FoxP3+ CD4+ T cells and RORɣt+ CD4+ T cells.CD4+ T cells were isolated by CD4+ Tcell isolation kit II human of Miltenyi Biotec and stained with respective antibodies and evaluated in relapsing phase (n = 20) and remitting phase (n = 20) of MS patients and healthy controls (n = 10). A forward and side scatter gate was used to select lymphocyte population and fluorescence compensation was set according to labeled lymphocytes with only green and only red fluorescent separately versus isotype control (A). Percentage of RORγt+ CD4+ T cells measured by Flow cytometry, shows meaningful increase in relapsing group (B) while percentage of FoxP3+ CD4+ T cells elevates in remitting group (C) (*p < 0.05, **p < 0.01 and ***p < 0.005, non-parametric Mann-Whitney t-test) (RP: Relapsing patient, MP: Remitting patient, HV: Healthy volunteer).
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pone.0124555.g001: Flow cytometry of FoxP3+ CD4+ T cells and RORɣt+ CD4+ T cells.CD4+ T cells were isolated by CD4+ Tcell isolation kit II human of Miltenyi Biotec and stained with respective antibodies and evaluated in relapsing phase (n = 20) and remitting phase (n = 20) of MS patients and healthy controls (n = 10). A forward and side scatter gate was used to select lymphocyte population and fluorescence compensation was set according to labeled lymphocytes with only green and only red fluorescent separately versus isotype control (A). Percentage of RORγt+ CD4+ T cells measured by Flow cytometry, shows meaningful increase in relapsing group (B) while percentage of FoxP3+ CD4+ T cells elevates in remitting group (C) (*p < 0.05, **p < 0.01 and ***p < 0.005, non-parametric Mann-Whitney t-test) (RP: Relapsing patient, MP: Remitting patient, HV: Healthy volunteer).

Mentions: First, to evaluate the number of Th17 and Treg cells in different phases of MS, part of CD4+ T cells yielded from MACS were stained with FoxP3, RORγt and CD4 antibodies as explained in materials and methods. Results showed that percentage of RORγt+ CD4+ T cells significantly increased in relapsing versus remitting and control group (p value = 0.0002, p value = 0.0003 respectively), while percentage of FoxP3+ CD4+ T cells significantly increased in remitting compared to relapsing and healthy controls (p value = 0.003, p value = 0.001 respectively) (Fig 1A–1C).


miR-141 and miR-200a, Revelation of New Possible Players in Modulation of Th17/Treg Differentiation and Pathogenesis of Multiple Sclerosis.

Naghavian R, Ghaedi K, Kiani-Esfahani A, Ganjalikhani-Hakemi M, Etemadifar M, Nasr-Esfahani MH - PLoS ONE (2015)

Flow cytometry of FoxP3+ CD4+ T cells and RORɣt+ CD4+ T cells.CD4+ T cells were isolated by CD4+ Tcell isolation kit II human of Miltenyi Biotec and stained with respective antibodies and evaluated in relapsing phase (n = 20) and remitting phase (n = 20) of MS patients and healthy controls (n = 10). A forward and side scatter gate was used to select lymphocyte population and fluorescence compensation was set according to labeled lymphocytes with only green and only red fluorescent separately versus isotype control (A). Percentage of RORγt+ CD4+ T cells measured by Flow cytometry, shows meaningful increase in relapsing group (B) while percentage of FoxP3+ CD4+ T cells elevates in remitting group (C) (*p < 0.05, **p < 0.01 and ***p < 0.005, non-parametric Mann-Whitney t-test) (RP: Relapsing patient, MP: Remitting patient, HV: Healthy volunteer).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4418573&req=5

pone.0124555.g001: Flow cytometry of FoxP3+ CD4+ T cells and RORɣt+ CD4+ T cells.CD4+ T cells were isolated by CD4+ Tcell isolation kit II human of Miltenyi Biotec and stained with respective antibodies and evaluated in relapsing phase (n = 20) and remitting phase (n = 20) of MS patients and healthy controls (n = 10). A forward and side scatter gate was used to select lymphocyte population and fluorescence compensation was set according to labeled lymphocytes with only green and only red fluorescent separately versus isotype control (A). Percentage of RORγt+ CD4+ T cells measured by Flow cytometry, shows meaningful increase in relapsing group (B) while percentage of FoxP3+ CD4+ T cells elevates in remitting group (C) (*p < 0.05, **p < 0.01 and ***p < 0.005, non-parametric Mann-Whitney t-test) (RP: Relapsing patient, MP: Remitting patient, HV: Healthy volunteer).
Mentions: First, to evaluate the number of Th17 and Treg cells in different phases of MS, part of CD4+ T cells yielded from MACS were stained with FoxP3, RORγt and CD4 antibodies as explained in materials and methods. Results showed that percentage of RORγt+ CD4+ T cells significantly increased in relapsing versus remitting and control group (p value = 0.0002, p value = 0.0003 respectively), while percentage of FoxP3+ CD4+ T cells significantly increased in remitting compared to relapsing and healthy controls (p value = 0.003, p value = 0.001 respectively) (Fig 1A–1C).

Bottom Line: Expression level of miR-141 and miR-200a were measured by RT-q PCR and compared to healthy control group (n=10).Furthermore, both miR-141 and miR-200a show up-regulation in relapsing phase of MS patients compared to remitting and control groups.Our data suggest that these miRNAs may probably inhibit negative regulators of Th17 cell differentiation, thus promoting its differentiation.

View Article: PubMed Central - PubMed

Affiliation: Division of Cellular and Molecular Biology, Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran; Department of Cellular Biotechnology at Cell Science research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.

ABSTRACT

Background: One of the main issues in pathogenesis of MS is Th17/Treg imbalance. There are growing interests in nominating miRNAs involved in Th17 cell differentiation, suggesting them as new therapeutic agents that may reduce progression of different autoimmune diseases specially MS.

Objectives: We assessed transcript levels of miR-141 and miR-200a in MS patients, during relapsing and remitting phases. We also investigated possible role of miR-141, miR-200a in inducing differentiation to Th17 cells.

Materials and methods: Forty RR-MS patient samples including relapsing (n=20) and remitting (n=20) phases were chosen. Expression level of miR-141 and miR-200a were measured by RT-q PCR and compared to healthy control group (n=10). In-silico analyses on miR-141 and miR-200a targetome showed involvement of both miRNAs in T helper cell differentiation pathways including TGF-β, mTOR and JAK/STAT.

Results: We observed that percentage of RORγt+ CD4+ T cells increase in relapsing phase while FOXP3+ CD4+ increase in remitting phase of MS patients. Furthermore, both miR-141 and miR-200a show up-regulation in relapsing phase of MS patients compared to remitting and control groups. Interestingly, expression level of target genes of miR-141 and miR-200a, which were assessed through in-silico methods, show down-regulation in relapsing phase of MS patients.

Conclusions: According to our results, miR-141 and miR-200a may be key miRNAs in progression of symptoms of MS through inducing differentiation of Th17 cells and inhibiting differentiation to Treg cells. Our data suggest that these miRNAs may probably inhibit negative regulators of Th17 cell differentiation, thus promoting its differentiation.

No MeSH data available.


Related in: MedlinePlus