Figure 6.

iv8-binding to human peripheral blood cells. Single naive human B cells were purified by cell sorting with fluorescently labeled iv8, and the IgL kappa transcripts were recovered by RT-PCR and sequenced or subjected to paired BCR sequencing using the chromium platform from 10X Genomics. As a control, naive human B cells were enriched using magnetic beads, and paired BCR sequencing was performed using the chromium platform. (A) Frequency of IgL kappa transcripts derived from VK3-11 in iv8⁺ or total B cells. Each dot represents one iv8⁺ sort (n = 13) or paired chromium sequencing run of magnetic bead–enriched (total naive) B cells (n = 7). (B) Frequency sequenced IgL kappa transcripts with a 5-aa CDLR3 in iv8⁺ or total B cells. Each dot represents one iv8⁺ sort (n = 13) or chromium sequencing run of magnetic bead–enriched (total naive) B cells (n = 7). Lines in A and B indicate arithmetic means, and error bars indicate SEM. (C) IgVK gene usage for of iv8⁺ B cells with 5-aa CDRL3s identified in B. Number in the middle of the pie chart indicates the total number of IgL kappa sequences analyzed. Statistics were calculated using two-tailed unpaired Student’s t tests in A and B. **, P ≤ 0.01; ***, P ≤ 0.001. LC, light chain.