Extended Data Fig. 3: Blimp-1 + 1DER effector T cells migrate from medLN to the lung.
a-c, Histograms of transcription factor RORγt and T-bet from Teff population (live TCRβ + CD4 + CD45.1 + CD45.2+FoxP3- cells) in 1DER T cell isolated from medLN after 3d (n = 8) and 5d (n = 9) HDM. d-e, naïve CD451 + CD45.2 + 1DERYFP T cells were adoptive transferred into CD45.2+ recipients followed by total 3 (doses) immunization. FTY720 was given by i.p. 6 h after the 2nd dose of HDM and was given 6 h before the 3rd dose of HDM (n = 7). DMSO injection was as control group (n = 6). Representative flow cytometry and absolute cell number on YFP+ cells showing Blimp-1 expression from Teff population in the medLN (d) and lung (e). f, 1DER TCR Tg mice were crossed to IL-10 GFP mice (1DERGFP) and naïve 1DERGFP CD4 T cells (CD45.1/2+) were transferred into host (CD45.2+), followed by 3d of HDM immunization (i.n). Representative flow plots of IL-10GFP gated on 1DERGFP T cells and percentage of IL-10-expressing (GFP+) Teff and TFH cells in the medLN. n = 5. g, Sensitization and challenge protocol using HDM. Mouse lung tissue was harvested 3 min after antibody anti-Thy1.2 was injected by i.v. Tissue resident CD4 + T cells were identified by CD44+Thy1.2- (h-i) and Blimp-1 YFP+ cells (j) were identified on d30 from the lung (n = 3 for unimmunized group, n = 2 for HDM group). k-l, Representative flow cytometry of Blimp-1 expression from lung analyzed on d23 after immunization. Gated from TCRβ + CD4+FoxP3- effector cells. l, Frequency of Blimp-1 expression among effector cells. n = 2 in Blimp-1CD4Cre, n = 12 in iBlimp-1Δ/Δ (day-4 ~ 0), n = 7 in iBlimp-1Δ/Δ (day-1 ~ 3), n = 12 in iBlimp-1Δ/Δ(day1 ~ 5), n = 7 in iBlimp-1Δ/Δ(day6 ~ 10), n = 5 in iBlimp-1Δ/Δ(day11 ~ 15), n = 20 in iBlimp-1+/+, n = 9 in Blimp-1+/++vehicle, n = 9 in Blimp-1Δ/Δ +vehicle. Each point represents one individual sample. Data are shown as means ± SD above and presented one independent experiment (j), two independent experiments (b-f) or three independent experiments (l). Two-tailed unpaired Mann-Whitney t test (d-f), Kruskal-Wallis one-way ANOVA test (l). d: **p = 0.0023, e: ***P = 0.006; f: **p = 0.0079; l: *p < 0.036, **p < 0.0079.
