Zhang, R., P. T. Sage, K. Finn, A. Huynh, B. R. Blazar, F. Marangoni, T. R. Mempel, A. H. Sharpe, and L. A. Turka. 2017. B cells drive autoimmunity in mice with CD28-deficient regulatory T cells. J. Immunol. 199: 3972–3980.

Panels (E)–(H) were inadvertently omitted from Fig. 3 in the published article. The figure legend was correct as published and is shown below for reference. Fig. 3 has been corrected in the online version of the article, which now differs from the print version.

FIGURE 3.

CD28 is required for TFR cell differentiation in LNs. (A) Percentage and number of TFH in total CD4+YFP cells and percentage and number of TFR in CD4+YFP+ Tregs in LNs of WT and CD28-ΔTreg mice (YFP is a Foxp3 reporter). (B) WT and CD28-ΔTreg mice were s.c. immunized with NP-OVA/CFA. After 7 d, TFH and TFR percentage were analyzed as in (A). Representative flow cytometry plots (left) and quantification (right). (C and D) In healthy CD28fl/fl × Foxp3YFP-CRE/+ female mice (unimmunized or NP-OVA immunized), CD28-deficient TFR (CD4+ Foxp3+YFP+CXCR5+ICOS+), and CD28-sufficient TFR (CD4+Foxp3+YFPCXCR5+ICOS+) were gated as (C) (representative of immunized dLN), and the percentage of TFR were shown in (D). Two to three animals were analyzed. (EH) Splenocytes of WT and CD28-ΔTreg mice were stimulated in vitro with PMA and ionomycin for 5 h in the presence of Golgi block. Representative flow cytometry of analyses of IL-4, IL-17, IL-21, and IFN-γ in CD4+ cells (E) and IFN-γ in CD8+ cells (G) and their quantification (F and H) are shown. Data are representative of over four WT and four CD28-ΔTreg mice analyzed. *p < 0.05, **p < 0.01, ***p < 0.001.

FIGURE 3.

CD28 is required for TFR cell differentiation in LNs. (A) Percentage and number of TFH in total CD4+YFP cells and percentage and number of TFR in CD4+YFP+ Tregs in LNs of WT and CD28-ΔTreg mice (YFP is a Foxp3 reporter). (B) WT and CD28-ΔTreg mice were s.c. immunized with NP-OVA/CFA. After 7 d, TFH and TFR percentage were analyzed as in (A). Representative flow cytometry plots (left) and quantification (right). (C and D) In healthy CD28fl/fl × Foxp3YFP-CRE/+ female mice (unimmunized or NP-OVA immunized), CD28-deficient TFR (CD4+ Foxp3+YFP+CXCR5+ICOS+), and CD28-sufficient TFR (CD4+Foxp3+YFPCXCR5+ICOS+) were gated as (C) (representative of immunized dLN), and the percentage of TFR were shown in (D). Two to three animals were analyzed. (EH) Splenocytes of WT and CD28-ΔTreg mice were stimulated in vitro with PMA and ionomycin for 5 h in the presence of Golgi block. Representative flow cytometry of analyses of IL-4, IL-17, IL-21, and IFN-γ in CD4+ cells (E) and IFN-γ in CD8+ cells (G) and their quantification (F and H) are shown. Data are representative of over four WT and four CD28-ΔTreg mice analyzed. *p < 0.05, **p < 0.01, ***p < 0.001.

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