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. 2023 Feb 2;108(6):1579–1589. doi: 10.3324/haematol.2022.281762

Figure 3.

Figure 3.

Recombinant FVIII co-stimulation with lipopolysaccharide plus plasma mainly induces proliferation of central memory T cells. Dendritic cells (DC) from HLA-DRB1*11-positive donors were treated with recombinant (r)FVIII (1 IU/mL), lipopolysaccharide (LPS) (0.1 mg/mL), and plasma (2.5 mL/mL). After 24 hours, carboxyfluorescein succinimidyl ester (CFSE)-labeled autologous CD4+ T cells were added. On day 9 of co-culture, T cells were harvested and stained with HLA-matched FVIII-specific tetramer and an antibody mix containing anti-CD3, anti-CD4, anti-CD45RA, anti-CD45RO, and anti-CCR7 antibodies. The CD4+ T cells were defined as 4 subsets: central memory (CM) T cells (CD45RO+CD45RA-CCR7+), effector memory (EM) T cells (CD45RO+CD45RA-CCR7-), naïve T cells (CD45RO-CD45RA+CCR7+), and effector T cells (TE) (CD45RO-CD45RA+CCR7-). (A) Data from 1 representative donor are shown. (B) Percentage distribution of CD4+ T-cell subtypes from proliferated CD4+ T cells (n=17, 5 donors from 7 independent experiments) and (C) from proliferated CD4+ FVIII-specific T cells (n=7, 6 donors from 2 independent experiments) are summarized. Error bars indicate the mean ± standard error of the mean. SSC: side scatter; FSC: forward scatter.