FIG 6.

Identification of Tpp17-specific rabbit IgG⁺ B cells. (A) Gating strategy: After gating lymphocytes, doublets were excluded in FSC-H and -W, and SSC-H and -W plots. IgG⁺ cells were identified from the IgM and IgA double-negative cells. (B) Tpp17-specific B cells were gated from IgG⁺ cells as double-positive cells for Tpp17-SP-AF405 and Tpp17-SP-AF647. B. burgdorferi OspC conjugated to SP-AF405 and SP-AF647 was used as a negative control. The mean frequencies of antigen-specific IgG⁺ B cells are shown in the accompanying bar graphs. (C) Identification of IgG⁺ B cells specific for PfTrx^BamA/ECL4 using the gating strategy shown in panel A. IgG⁺ PfTrx⁺ cells were excluded using PfTrx-SP-APC-Cy7. From each immune rabbit, B cells specific for PfTrx^BamA/ECL4 were identified within the IgG⁺ PfTrx^Neg gate as cells double-positive for SP-AF405 and SP-AF647; PBMCs from a normal rabbit were used as a control. The mean frequencies of IgG⁺ B cells specific BamA ECL4 are shown in the accompanying bar graphs.