Figure 6.

The Tfh-dependent TLS predicts tumor prognosis of patients with early-stage LUAD and inhibits tumor growth

(A) Kaplan-Meier curves showing the overall survival values of activated memory B (left) and GC B (right) cell scores in TCGA database (STAR Methods).

(B) Kaplan-Meier curves showing the overall survival values of Tfh-like cell signature scores in TCGA database.

(C) Quantification of CD20⁺ cells, density of TLSs, and ratio of tumor area occupied by TLSs in TLS^high and TLS^low patients. TLS^high, n = 27; TLS^low, n = 15. Bars and error bars represent mean +/- standard error.

(D) Kaplan–Meier estimates of overall survival shown by in TLS^high and TLS^low patients. TLS^high, n = 27; TLS^low, n = 15.

(E) Ratio of tumors to mm² of the lung tissues obtained from anti-CD20-Ab-treated KPC mice. Control (n = 6) and anti-CD20-treated KPC mice (n = 6).

(F) Multiplex immunostaining assay of TLSs for the following markers: CD3, CD19, and DAPI in anti-C20-treated KPC mice. Scale bars, 50 μm. Control (n = 6) and anti-CD20-treated KPC mice (n = 6).

(G) Schematic illustration showing IL-21R Ab neutralization of control and tumor-bearing CD4^CRE; Bcl6^fl/fl mice.

(H and I) LLC cells were intraperitoneally injected into 8-week-old controls, CD4^CRE; Bcl6^fl/fl mice, control+IL-21R Ab and CD4^CRE; Bcl6^fl/fl+IL-21R Ab mice. The image and weight of tumors were obtained at 16 days post-tumor inoculation. Control (n = 5), CD4^CRE; Bcl6^fl/fl (n = 5), control+IL-21R Ab (n = 5), and CD4^CRE; Bcl6^fl/fl+ IL-21R Ab mice (n = 5). ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001, two-sided Mann-Whitney U test. Error bars indicate mean ± SEM.

(J) Multiplex immunostaining assay of TLS for the following markers: CD3, CD19, and DAPI in controls, CD4^CRE; Bcl6^fl/fl mice, control+IL-21R Ab, and CD4^CRE; Bcl6^fl/fl+IL-21R Ab mice. Scale bars, 50 μm.

(K and L) Density of TLSs and ratio of tumor area occupied by controls, CD4^CRE; Bcl6^fl/fl mice, control+IL-21R Ab, and CD4^CRE; Bcl6^fl/fl+IL-21R Ab mice. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001, two-sided Mann-Whitney U test. Error bars indicate mean ± SEM.