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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

Percentages of the induced CD4+CD25+ T cells generated from CD4 + CD25ˉ T cells by IFN-γ with different concentrations in MG patients. CD4+CD25ˉT cells were stimulated by IFN-γ with different concentrations (0, 20, 40, 80 ng/ml) in the presence of anti-CD3 (1 μg/ml) and anti-CD28 (1 μg/ml) antibodies for 72 h and then sorted to measure the percentages of induced CD4+CD25+ T cells by flow cytometry. The data are from one experiment and are representative of 19 separate experiments
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Fig3: Percentages of the induced CD4+CD25+ T cells generated from CD4 + CD25ˉ T cells by IFN-γ with different concentrations in MG patients. CD4+CD25ˉT cells were stimulated by IFN-γ with different concentrations (0, 20, 40, 80 ng/ml) in the presence of anti-CD3 (1 μg/ml) and anti-CD28 (1 μg/ml) antibodies for 72 h and then sorted to measure the percentages of induced CD4+CD25+ T cells by flow cytometry. The data are from one experiment and are representative of 19 separate experiments

Mentions: The conversion rate of the induced CD4+CD25+ T cells generated from CD4+CD25− T cells changes with the concentration of IFN-γ in MG patients. The percentage of the induced CD4+CD25+ T cells firstly increases and later declines with increasing concentration of IFN-γ (P < 0.05). In the presence of anti-CD3 and anti-CD28 antibodies without IFN-γ, the induced CD4+CD25+ T cells can be generated from CD4+CD25− T cells and the percentage of the induced CD4+CD25+ T cells is 18.3 ± 4.1 %. When the concentration of IFN-γ reaches as high as 40 ng/ml, the percentage of the induced CD4+CD25+ T cells has the highest value (43.1 ± 3.7 %). However, when the concentration of IFN-γ becomes higher as 80 ng/ml, the percentage of the induced CD4+CD25+ T cells decreases (29.8 ± 2.9 %) (Table 2). Figure 3 shows the percentages of the induced CD4+CD25+ T cells generated from CD4+CD25− T cells by IFN-γ with different concentrations in MG patients. It is from one typical experiment result representing the total 19 separate experiments.Table 2


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

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

Percentages of the induced CD4+CD25+ T cells generated from CD4 + CD25ˉ T cells by IFN-γ with different concentrations in MG patients. CD4+CD25ˉT cells were stimulated by IFN-γ with different concentrations (0, 20, 40, 80 ng/ml) in the presence of anti-CD3 (1 μg/ml) and anti-CD28 (1 μg/ml) antibodies for 72 h and then sorted to measure the percentages of induced CD4+CD25+ T cells by flow cytometry. The data are from one experiment and are representative of 19 separate experiments
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig3: Percentages of the induced CD4+CD25+ T cells generated from CD4 + CD25ˉ T cells by IFN-γ with different concentrations in MG patients. CD4+CD25ˉT cells were stimulated by IFN-γ with different concentrations (0, 20, 40, 80 ng/ml) in the presence of anti-CD3 (1 μg/ml) and anti-CD28 (1 μg/ml) antibodies for 72 h and then sorted to measure the percentages of induced CD4+CD25+ T cells by flow cytometry. The data are from one experiment and are representative of 19 separate experiments
Mentions: The conversion rate of the induced CD4+CD25+ T cells generated from CD4+CD25− T cells changes with the concentration of IFN-γ in MG patients. The percentage of the induced CD4+CD25+ T cells firstly increases and later declines with increasing concentration of IFN-γ (P < 0.05). In the presence of anti-CD3 and anti-CD28 antibodies without IFN-γ, the induced CD4+CD25+ T cells can be generated from CD4+CD25− T cells and the percentage of the induced CD4+CD25+ T cells is 18.3 ± 4.1 %. When the concentration of IFN-γ reaches as high as 40 ng/ml, the percentage of the induced CD4+CD25+ T cells has the highest value (43.1 ± 3.7 %). However, when the concentration of IFN-γ becomes higher as 80 ng/ml, the percentage of the induced CD4+CD25+ T cells decreases (29.8 ± 2.9 %) (Table 2). Figure 3 shows the percentages of the induced CD4+CD25+ T cells generated from CD4+CD25− T cells by IFN-γ with different concentrations in MG patients. It is from one typical experiment result representing the total 19 separate experiments.Table 2

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