Fig. 5: THX mice develop thymic huTECs and huB cells, differentiate huT_FH cells, form GCs and generate class-switched huMBCs.

a–k, THX (n = 5), huNBSGW (n = 7) and JAX NSG huCD34 (n = 3) mice of the n = 7, 7 and 4 mice, respectively, of Fig. 4, were injected i.p. with NP₁₆-CGG on day 0, boosted on day 14 and euthanized on day 28, unless otherwise specified. a, Spleen and LNs (cervical, mesenteric, mediastinal and axillary) of NP₁₆-CGG-immunized THX, huNBSGW and JAX NSG huCD34 mice and non-immunized NSG and C57BL/6 mice (scale bar, 1 cm). b, Spleen huB cells, huT cells, huNK cells, huDCs and human monocytes in THX versus huNBSGW and JAX NSG huCD34 mice (39.5 ± 6.6 × 10⁶ versus 26.0 ± 4.4 × 10⁶ and 11.9 ± 1.5 × 10⁶ mononuclear cells). c, Blood and spleen huCD19⁺CD5⁺ (B1) cells in THX, huNBSGW and JAX NSG huCD34 mice on days 0 and 28. d,e, Mesenteric LN GC huCD27⁻CD20⁺CD38⁺ B cells (d, day 14) and blood class-switched memory huCD19⁺CD27⁺IgD⁻ B cells (e, day 90) in THX and huNBSGW mice. f, Spleen sections from THX, huNBSGW and JAX NSG huCD34 mice (day 14, hematoxylin and eosin (H&E) and immunohistochemical huCD20, huCD3, huKi67, huBCL6, huAID and huBLIMP-1; scale bars, 1.0 mm and 100 μm). g,h, huCD4⁺, huCD8⁺, huCD4⁺CD8⁺ T cells (g) and huCD4⁺CXCR5⁺PD-1⁺ T_FH cells (h) in spleens and mesenteric LNs of THX, huNBSGW and JAX NSG huCD34 mice. i, Thymus sections of THX and huNBSGW mice (whole organ, H&E, human and mouse EpCAM⁺ TECs immunofluorescence; scale bars, 100 μm, 400 µm and 5 mm). j,k, Human and mouse EpCAM⁺ TECs, huT cells, huB cells, huDCs, human monocytes (j) and cells expressing huMHC class I and huMHC class II (k) in THX mouse thymus. Images (a), micrographs (f and i) and FACS plots (c–e, g, h, j and k) are from one mouse per group, each representative of three mice. huCD45⁺ cells were pre-gated in all FACS analyses. In the histograms (b, g and h), each dot represents an individual mouse and bars depict the mean with s.e.m. Statistical significance (b, g and h) was assessed by two-sided Student’s unpaired t-test (NS, not significant; *P < 0.05, **P < 0.01, ***P < 0.001).

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