cd45ra macs  (Miltenyi Biotec)

Journal: Endocrine Connections

Article Title: Elevated thyroglobulin level is associated with dysfunction of regulatory T cells in patients with thyroid nodules

doi: 10.1530/EC-18-0545

Figure Lengend Snippet: The relationship between blood Tg levels and Treg cell function. (A) Blood Tg concentrations in 30 patients with benign thyroid nodules were measured. Tregs from these patients were co-cultured with CD4 + CD25 − T cells (Teffs) at a 1:10 ratio and their immunomodulatory function were determined. The inhibitory function of Tregs were analyzed using the inhibition rate (%) of Teffs proliferation and was calculated using the equation: ((proliferation of Teffs − proliferation of Teffs plus Treg)/proliferation rate of Teffs alone) × 100%. The correlation between Tg concentration and the inhibition rate (%) of Teffs proliferation (representing Treg cell function) was analyzed by linear regression. (B) Patients were divided into high Tg group (Tg >77 μg/L) and normal group (Tg ≤77 μg/L). Tregs function were compared between the two groups. * P < 0.05.

Article Snippet: Tregs were isolated from mononuclear cells using a human Treg cell magnetic-activated cell sorting (MACS) kit (Miltenyi Biotec).

Techniques: Cell Function Assay, Cell Culture, Inhibition, Concentration Assay

Journal: Endocrine Connections

Article Title: Elevated thyroglobulin level is associated with dysfunction of regulatory T cells in patients with thyroid nodules

doi: 10.1530/EC-18-0545

Figure Lengend Snippet: Suppressive functionality of thyroid Tregs. (A) Thyroid tissues were taken from six patients who had undergone surgery for benign thyroid nodules. Thyroid Tregs were co-cultured with peripheral blood effector T cells (Teffs, CD4 + CD25 − T cells) at different ratios (1:2, 1:4, and 1:8) for 3 days. Teffs were labeled with CFSE and analyzed by flow cytometry. (B) The inhibitory function of Tregs were analyzed using the inhibition rate (%) of Teffs proliferation and was calculated using the equation: ((proliferation of Teffs − proliferation of Teffs plus Treg)/proliferation rate of Teffs alone) × 100%. The experiment was performed independently six times.

Article Snippet: Tregs were isolated from mononuclear cells using a human Treg cell magnetic-activated cell sorting (MACS) kit (Miltenyi Biotec).

Techniques: Cell Culture, Labeling, Flow Cytometry, Inhibition

Journal: Endocrine Connections

Article Title: Elevated thyroglobulin level is associated with dysfunction of regulatory T cells in patients with thyroid nodules

doi: 10.1530/EC-18-0545

Figure Lengend Snippet: The bidirectional effects of Tg on Treg cell function. (A) Tregs were isolated from peripheral blood of healthy donors and co-cultured with CD4 + CD25 − T cells labeled with CFSE at different ratios (1:2, 1:4 and 1:8), and simultaneously treated with TG at various concentrations (0, 0.1, 1, 10 and 100 μg/mL) for 3 days. Anti-human CD3 (5 × 10 −8 mol/L) and anti-human CD28 (2.27 × 10 −8 mol/L) antibodies were also added to the cultures. The proliferation rates of Teffs were analyzed by flow cytometry. (B) Inhibitory function of Tregs, represented by the inhibition rate (%) on Teffs proliferation, was calculated using the equation: ((proliferation of Teffs − proliferation of Teffs plus Treg)/proliferation rate of Teffs alone) × 100%. The experiment was performed independently 11 times. * P < 0.05, ** P < 0.01.

Article Snippet: Tregs were isolated from mononuclear cells using a human Treg cell magnetic-activated cell sorting (MACS) kit (Miltenyi Biotec).

Techniques: Cell Function Assay, Isolation, Cell Culture, Labeling, Flow Cytometry, Inhibition