CD71+VISTA+ erythroid cells promote the development and function of regulatory T cells through TGF-β
Fig 6
Newly generated CD71+ erythroid cells compensate for the loss of their counterparts.
(A) Representative flow dot plots illustrating the frequency of CD71+ erythroid cells in the spleen of isotype (Rat-IgG) versus anti-CD71–treated newborn mice. (B) The percentage of CD71+ erythroid cells in the spleen of anti-CD71–treated compared to control groups in 9-day-old BALB/c mice. (C) Representative dot plots illustrating the percentages of CD4+FOXP3+ in the spleen of isotype versus anti-CD71–treated mouse. (D) Percentage of CD4+FOXP3+ in the spleen of anti-CD71–treated compared to control groups in 9-day-old BALB/c mice. (E) Gene expression of TGF-β–associated genes, galectins, and PDL-1 (CD274) in enriched CD71+ erythroid cells from anti-CD71–treated versus controls. (F) Representative histogram of VISTA expression on CD71+ erythroid cells of control versus anti-CD71–treated mouse. (G) Percentages of CD71+VISTA+ erythroid cells among splenocytes of control and anti-CD71–treated BALB/c mice. (H) The MFI of VISTA on CD71+ erythroid cells from the spleen of control versus anti-CD71–treated mice. (I) RNAseq data showing expression of TGF-β1 and (J) TGF-β2 mRNA in CD71+ erythroid cells obtained from control or anti-CD71–treated mice. (K) PCR data showing expression of TLR2, and (L) TLR4 genes in CD71+ erythroid cells obtained from control or treated with anti-CD71 antibody. (M) The representative zebra plots showing surface expression of TLR2 on CD71+ erythroid cells. (N) Cumulative data showing percentages of TLR+CD71+ erythroid cells in control versus anti-CD71–treated group. (O) Representative zebra plots showing percentage of TGF-β+ cells among CD71+ erythroid cells from controls or treated mice with anti-CD71 antibody. (P) Cumulative data showing percentages of TGF-β expressing CD71+ erythroid cells in rat-IgG–versus anti-CD71–treated mice. (Q) The expression of Id1, (R) Id2, (S) PDL-1, (T) Lgals3, and (U) Lgals1 mRNA levels are shown in CD71+ erythroid cells from either control or anti-CD71–treated mice using RNAseq. Each point represents data from an individual mouse, representative of at least three independent experiments. Bar, mean ± one standard error. The underlying data can be found in S1 Data. BALB/c, mouse strain; CD71, cell-surface transferrin receptor; FOXP3, forkhead box P3; Gal, Galectin; Gata1, erythroid transcription factor; ID, inhibitor of DNA binding; Ig, immunoglobulin; IgG, immunoglobulin G; Iso., isotype; Lgals, Galectin genes; MFI, mean fluorescence intensity; mRNA, messenger RNA; PDL-1, program death ligand-1; RNAseq, RNA sequencing; TGF-β, transforming growth factor beta; TLR, Toll-like receptor; VISTA, V-domain Ig Suppressor of T Cell Activation.