Cd19 clone 1d3 cd19

Mice were s.c. immunized with alum-adsorbed NP–OVA (50 µg) and TLR7–alum or TLR7-NP (equivalent gardiquimod dose, 20 µg) in 100 µl PBS at tail base. Inguinal dLNs were excised at day 4, day 7, day 14 and day 22 to prepare single-cell suspensions. For flow cytometry analysis of GC B cells, follicular T cells (TFH) and plasmablasts, cells from the dLNs were stained with Ghost Dye Violet 510 (Tonbo Biosciences). Cells were then washed and blocked with Fc-blocker (clone 2.4G2, BD Bioscience) prior to staining with markers, including CD19 (clone 1D3/CD19, Biolegend), CD38 (clone 90, BD Biosciences), CD95 (clone Jo2, BD Biosciences), CD138 (clone 281-2, BD Biosciences), CD44 (clone IM7, BioLegend), CD3 (clone 17A2, BioLegend), CD4 (clone GK1.5, BioLegend), CXCR5 (clone L138D7, BioLegend) and PD1 (clone 29F.1A12, BioLegend). After staining, cells were washed and fixed with 1.5% PFA. Stained cells were collected using BD FACS Diva v8.01 software associated with a BD LSRII flow cytometer. Data were analysed with FlowJo 10 software. The gating strategy consisted of gating GCBC on live single CD3⁻CD19⁺CD95⁺CD38⁻ cells, TFH on live single CD19⁻CD3⁺CD4⁺CXCR5⁺PD1^hi, and plasmablasts on live single CD138⁺CD44⁺ cells.

The cell suspension from cardiac tissue was prepared as described above. Cells were incubated with Fixable Viability Dye (eBioscience, San Diego, CA, USA, cat. no 65-0865-14, Thermo Fisher Scientific, Waltham, MA, USA). The cardiac inflammatory cell profile was evaluated based on the following markers: CD45 (clone 30-F11, cat. no 563891, BD Biosciences), CD19 (clone 1D3/CD19, no cat. 152404, BioLegend, San Diego, CA, USA), CD3 (clone 17A2, cat. no 100217, BioLegend), CD8a (clone 53-6.7, cat. no 100722, Biolegend), CD4 (clone RM4-5, cat. no 100530, BioLegend), CD25 (clone 3C7, cat. no 101904, BioLegend), and Ly6G (clone 1A8, cat. no 127628, BioLegend). The stained cells were washed, suspended in PBS, and analyzed using flow cytometry (FACSCanto II, Becton-Dickinson).