Fig. 4.

Recombinant leucine-rich alpha 2 glycoprotein (LRG) enhances differentiation of T helper 17 (Th17) cells under the Th17 polarization condition. a Naïve CD4 T cells (n = 3) were treated for 3 days with anti-CD3/CD28 beads and transforming growth factor beta (TGF-β) (0, 0.125 or 0.5 ng/mL (Treg polarization condition)) in the presence or absence of LRG (LRG + (1 μg/mL) or LRG -). The percentage of CD4⁺Foxp3⁺ T regulatory (Treg) cells was determined by flow cytometry (right): *p < 0.05. Representative data are shown (left panel). b, c, d Naïve CD4 T cells were isolated from the spleen and lymph nodes of LRG knockout mice (LRG KO) (n = 3) and cultured for 3 days with anti-CD3/CD28 beads in the presence or absence of recombinant LRG (LRG + (1 μg/mL) or LRG -) under a non-polarization condition without cytokines or the Th17 polarization condition with TGF-β (2 ng/mL) and IL-6 (100 ng/mL). b The expression of IL-17 (left) and RORγt (right) mRNA was analyzed by quantitative PCR: *p < 0.05. c The percentage of IL-17+ cells in the CD4+ population under the Th17 polarization condition was determined by flow cytometry. Values are mean ± SD, n = 3 (left panel). Representative data are shown (right panel). d The induction of Treg cells under a non-polarization condition or under the Th17 polarization condition was examined by flow cytometry. All statistical analyses were performed using Student’s t test. NT; non-treated