Fig. 3

CD20⁺ B cells are associated with good prognosis of NSCLC. a, b Validation of the presence of CD79A⁺CD20⁺ and CD79A⁺CD20⁻ B cells in NSCLC tumors. Antibodies against CD79A and CD20 were used for immunofluorescence and immunohistochemistry. a Co-staining of CD79A (red) and CD20 (green), as well as the staining of CD79A only, was illustrated by immunofluorescence. b Immunohistochemistry showing the distribution of CD79A and CD20 in NSCLC tissue. The CD20⁺ B cells were located at the tertiary lymphoid structures (TLS) only, but the CD79⁺ B cells were located at both TLS and the tumor regions. c, d Flow cytometry showing the CD79A⁺CD20⁺ cells were reduced in advanced stages of NSCLC. c Representative illustration of flow cytometry results was plotted. d The ratio of CD79A⁺CD20⁺ and CD79A⁺CD20⁻ in 30 tumor tissues from I–III stages of NSCLC were plotted. e, f Patient with high levels of infiltration of naïve-like B cells was associated with better clinical outcome of NSCLC. e The naïve-like B score determined by immunohistochemistry in I–III stages of NSCLC was plotted. f The enrichment of naïve-like B cells (Naïve-like B^high) was correlated with good prognosis of NSCLC in 164 patients. The difference in overall survival and relapse-free-survival between Naïve-like B^high patients and Naïve-like B^low patients was determined by log-rank test. The NSCLC tumors were divided into two subgroups, including Naïve-like B^high and Naïve-like B^low according to the immunostaining of CD79A and CD20 across 164 NSCLC tissues. g Validation using TCGA datasets. The mRNA level of MS4A1(CD20) in different stages of NSCLC as well as normal control was plotted. MS4A1(CD20) was highly expressed in lung cancer as compared to adjacent normal tissues but decreased in advanced stages of lung cancers in the TCGA cohort. h, i Correlation between MS4A1(CD20) expression and PDCD1 (PD-1)/CD274 (PD-L1) as well as tumor mutation burden of LUAD. The expression of MS4A1 (CD20) and PDCD1/CD274 was obtained from the TCGA-LUAD cohorts. The tumor mutation burden (TMB) was calculated and divided into the high TMB and low TMB groups