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. 2017 May 19;198(12):4629–4638. doi: 10.4049/jimmunol.1600773

FIGURE 3.

FIGURE 3.

Identification and phenotyping of AMA1-specific B cells with an AMA1 tetramer in human samples. Representative human flow plots. B cells were identified by flow cytometry in PBMCs. (A) Single cells are selected using side scatter area and width. (B) Lymphocytes were selected based on side and forward scatter. (C) B cells were selected by choosing cells that expressed CD19 and did not express CD14, CD16, CD4, or CD8 (Dump). (D) Mature B cells were selected based on expression of CD10. (E1 and E2) Among mature B cells, naive (CD21hi,CD27−), resting memory (CD21hi,CD27+), activated and plasma cells/blasts (CD21low, CD27+), and atypical (CD21low,CD27−) memory B cells were identified based on CD27 and CD21 expression. (F1 and F2) Among mature B cells, AMA1 and decoy-specific cells are identified using the tetramer and AF680-PE labeled streptavidin. (G1 and G2). Among AMA1-specific B cells, naive (CD21hi,CD27−), resting memory (CD21hi,CD27+), activated and plasma cells/blasts (CD21low, CD27+) and atypical (CD21low,CD27−) memory B cells were identified based on CD27 and CD21 expression, using the same gates as applied to the whole mature B cell population. Numbers indicate percentage of total number of events on that graph that fall within gate.