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Feasibility of up-regulating CD4(+)CD25(+) Tregs by IFN-γ in myasthenia gravis patients.

Huang S, Wang W, Chi L - BMC Neurol (2015)

Bottom Line: It shows the percentages of CD4(+)CD25(+) T cells among CD4(+) T cells have no significant difference in MG patients compared with those in HCs.This subject will further reveal the role of IFN-γ in the pathogenesis of MG from a new perspective.It will also provide the scientific basis for the clinical targeted therapy of MG.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, 150080, P.R. China. cecile_huang@163.com.

ABSTRACT

Background: In myasthenia gravis (MG) patients, the dysfunction of CD4(+)CD25(+) regulatory T cells (CD4(+)CD25(+) Tregs) may be one of the important pathogenesis of MG. Currently, the role of IFN-γ in autoimmune diseases is still controversial and needs further exploration. In this study, whether IFN-γ can induce CD4(+)CD25(-) T cells into CD4(+)CD25(+) Tregs in MG in vitro was investigated systematically.

Methods: Flow cytometry was used to analyze the number of CD4(+)CD25(+) Tregs in MG patients and healthy controls (HCs). CD4(+)CD25(-) T cells were separated from the peripheral blood mononuclear cells of MG patients and HCs, and the CD4(+)CD25(+) Tregs were separated from HCs by Magnetic cell sorting (MACS). IFN-γ with different concentrations was used to stimulate CD4(+)CD25(-) T cells. The percentages of the induced CD4(+)CD25(+) T cells were detected by flow cytometry. The FoxP3 expression of the induced CD4(+)CD25(+) T cells in MG patients was detected by real-time PCR at mRNA level. The induced CD4(+)CD25(+) T cells were co-cultured with autologous CD4(+)CD25(-) T cells to estimate the suppressive ability of the induced CD4(+)CD25(+) T cells to CD4(+)CD25(-) T cells.

Results: It shows the percentages of CD4(+)CD25(+) T cells among CD4(+) T cells have no significant difference in MG patients compared with those in HCs. There is also merely no difference in the percentages of CD4(+)CD25(+) T cells between thymectomized and non-thymectomized MG patients. CD4(+)CD25(-) T cells can be induced to CD4(+)CD25(+) T cells after applying IFN-γ in MG patients and HCs. The proportion and FoxP3 expression of the induced CD4(+)CD25(+) T cells are the highest at the level of 40 ng/ml IFN-γ, and the suppressive function of the CD4(+)CD25(+) T cells induced by 40 ng/ml IFN-γ is the strongest in MG patients.

Conclusions: This subject will further reveal the role of IFN-γ in the pathogenesis of MG from a new perspective. It will also provide the scientific basis for the clinical targeted therapy of MG.

No MeSH data available.


Related in: MedlinePlus

a Comparation of the percentages of CD4+CD25+ Tregs between MG patients and healthy controls (HCs). b Percentages of CD4+CD25+ T cells in MG patients and HCs. a MNCs are stained with PerCP-CD4 and PE-CD25. Gate was set on CD4+ T cell population. The data are from one experiment and are representative of 17 separate experiments. b The percentage of CD4+CD25+ T cells among CD4+ T cells in peripheral blood from MG patients is similar to that from HCs
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Fig1: a Comparation of the percentages of CD4+CD25+ Tregs between MG patients and healthy controls (HCs). b Percentages of CD4+CD25+ T cells in MG patients and HCs. a MNCs are stained with PerCP-CD4 and PE-CD25. Gate was set on CD4+ T cell population. The data are from one experiment and are representative of 17 separate experiments. b The percentage of CD4+CD25+ T cells among CD4+ T cells in peripheral blood from MG patients is similar to that from HCs

Mentions: In our experiment, Tregs were identified as CD4+CD25high T cells by selecting those CD4+ cells whose CD25 expression exceeded the level of CD25 positivity observed on the CD4 negative population [19]. The number of CD4+CD25+ Tregs reservoir in peripheral blood was measured by flow cytometry. As shown in Fig. 1a, the percentages of CD4+CD25+ Tregs in peripheral blood from MG patients (9.85 ± 2.04 %) and HCs (8.67 ± 3.38 %) are similar (P > 0.05). Figure 1b is displayed as an example for a typical case study of one experiment representing the total 17 similar separate experiments.Fig. 1


Feasibility of up-regulating CD4(+)CD25(+) Tregs by IFN-γ in myasthenia gravis patients.

Huang S, Wang W, Chi L - BMC Neurol (2015)

a Comparation of the percentages of CD4+CD25+ Tregs between MG patients and healthy controls (HCs). b Percentages of CD4+CD25+ T cells in MG patients and HCs. a MNCs are stained with PerCP-CD4 and PE-CD25. Gate was set on CD4+ T cell population. The data are from one experiment and are representative of 17 separate experiments. b The percentage of CD4+CD25+ T cells among CD4+ T cells in peripheral blood from MG patients is similar to that from HCs
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4562356&req=5

Fig1: a Comparation of the percentages of CD4+CD25+ Tregs between MG patients and healthy controls (HCs). b Percentages of CD4+CD25+ T cells in MG patients and HCs. a MNCs are stained with PerCP-CD4 and PE-CD25. Gate was set on CD4+ T cell population. The data are from one experiment and are representative of 17 separate experiments. b The percentage of CD4+CD25+ T cells among CD4+ T cells in peripheral blood from MG patients is similar to that from HCs
Mentions: In our experiment, Tregs were identified as CD4+CD25high T cells by selecting those CD4+ cells whose CD25 expression exceeded the level of CD25 positivity observed on the CD4 negative population [19]. The number of CD4+CD25+ Tregs reservoir in peripheral blood was measured by flow cytometry. As shown in Fig. 1a, the percentages of CD4+CD25+ Tregs in peripheral blood from MG patients (9.85 ± 2.04 %) and HCs (8.67 ± 3.38 %) are similar (P > 0.05). Figure 1b is displayed as an example for a typical case study of one experiment representing the total 17 similar separate experiments.Fig. 1

Bottom Line: It shows the percentages of CD4(+)CD25(+) T cells among CD4(+) T cells have no significant difference in MG patients compared with those in HCs.This subject will further reveal the role of IFN-γ in the pathogenesis of MG from a new perspective.It will also provide the scientific basis for the clinical targeted therapy of MG.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, 150080, P.R. China. cecile_huang@163.com.

ABSTRACT

Background: In myasthenia gravis (MG) patients, the dysfunction of CD4(+)CD25(+) regulatory T cells (CD4(+)CD25(+) Tregs) may be one of the important pathogenesis of MG. Currently, the role of IFN-γ in autoimmune diseases is still controversial and needs further exploration. In this study, whether IFN-γ can induce CD4(+)CD25(-) T cells into CD4(+)CD25(+) Tregs in MG in vitro was investigated systematically.

Methods: Flow cytometry was used to analyze the number of CD4(+)CD25(+) Tregs in MG patients and healthy controls (HCs). CD4(+)CD25(-) T cells were separated from the peripheral blood mononuclear cells of MG patients and HCs, and the CD4(+)CD25(+) Tregs were separated from HCs by Magnetic cell sorting (MACS). IFN-γ with different concentrations was used to stimulate CD4(+)CD25(-) T cells. The percentages of the induced CD4(+)CD25(+) T cells were detected by flow cytometry. The FoxP3 expression of the induced CD4(+)CD25(+) T cells in MG patients was detected by real-time PCR at mRNA level. The induced CD4(+)CD25(+) T cells were co-cultured with autologous CD4(+)CD25(-) T cells to estimate the suppressive ability of the induced CD4(+)CD25(+) T cells to CD4(+)CD25(-) T cells.

Results: It shows the percentages of CD4(+)CD25(+) T cells among CD4(+) T cells have no significant difference in MG patients compared with those in HCs. There is also merely no difference in the percentages of CD4(+)CD25(+) T cells between thymectomized and non-thymectomized MG patients. CD4(+)CD25(-) T cells can be induced to CD4(+)CD25(+) T cells after applying IFN-γ in MG patients and HCs. The proportion and FoxP3 expression of the induced CD4(+)CD25(+) T cells are the highest at the level of 40 ng/ml IFN-γ, and the suppressive function of the CD4(+)CD25(+) T cells induced by 40 ng/ml IFN-γ is the strongest in MG patients.

Conclusions: This subject will further reveal the role of IFN-γ in the pathogenesis of MG from a new perspective. It will also provide the scientific basis for the clinical targeted therapy of MG.

No MeSH data available.


Related in: MedlinePlus