Fig. 1

DN B cells are expanded in NSCLC tumors. Single cell suspensions of NSCLC tumors and matched normal lung tissue samples were stained with APC-CD79A, PE-IgD and FITC-CD27. a Schema for the enumeration of CD27⁻IgD⁻ DN B cells by flow cytometry (1) debris is excluded based on FCS-A, SSC-A, (2) single cells are selected based on FSC-W and FCS-A (3) then the total B cell population is gated based on APC-CD79A expression (4). Finally, the percentage of DN B cells were defined by the absence of PE-IgD and FITC-CD27 expression. b Shown is the percentage of CD79A⁺CD27⁻IgD⁻ DN B cells present out of total live cell singlet population analyzed by flow cytometry of tumor versus normal lung for each patient studied. The cases are arranged in ascending order according to the percentage of DN B cells within the tumor. c Ratio of DN B cells was calculated by dividing the percentage of DN B cells found within the tumor by that found within the matched normal tissue for each patient. Values are arranged in ascending order. Paired students t-test was used to evaluate the statistical differences between tumor and matched normal control tissues. *p < 0.001