Extended Data Fig. 6: THX mice huB cells undergo CSR and differentiation as efficiently as huB cells from adult humans.

a–c, Naïve huIgM⁺IgD⁺ B cells isolated from blood of healthy humans (n = 3, HS 14, 15, 16) and spleens of non-intentionally immunized THX mice (n = 3, THX 442, 443, 444) were cultured for 120 h upon stimulation with: (a) CD154 (3 U/ml), huIL-2 (100 ng/ml), huIL-4 (20 ng/ml) and huIL-21 (50 ng/ml), (b) CpG ODN2395 (2.5 mg/ml), huIL-2, huIL-21, TGF-β (4 ng/ml) and retinoic acid (4 ng/ml), or (c) CpG ODN2395, huIL-2, huIL-4 and huIL-21. Identification of huIgM⁺, huIgD⁺, huIgG⁺, huIgA⁺ or huIgE⁺ B cells, huCD27⁺IgD^– class-switched memory-like B cells (huMB) and huCD27⁺CD38⁺ PBs by flow cytometry. huCD45⁺CD19⁺ cells were pre-gated in all FACS analyses. d, AICDA, PRDM1, V_HDJ_H-Cμ, V_HDJ_H-Cγ1, V_HDJ_H-Cα1 and V_HDJ_H-Cε transcript expression in HS and THX mice huB cell microcultures (n = 3 biological replicates for each different microculture), as measured by qPCR and normalized to HPRT1 expression (2^−ΔCt method). In histograms, each dot represents transcript expression from one human or one THX mouse huB cell microculture and the bar depicts the mean with s.e.m. Statistical significance (d) was assessed by two-sided Student’s unpaired t-test (NS, not significant).

Source data