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. 2016 Dec 12;213(13):3007–3024. doi: 10.1084/jem.20160712

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© 2016 Hayakawa et al.

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Figure 2. — CLL development from early generated B-1–derived B cells. (A) Comparison of CLL incidence in C.B17.scid recipients transferred with BM HSC, B1a, and non-B1a B cells purified from 2 mo TC⁺ C.B17 mice. pB1a and sB1a (each 5 × 10⁵/recipient), and non-B1a spleen B cells (sB; 3 × 10⁶/recipient) were cotransferred with TC^– BALB/c BM. 2 mo after transfer, PBL showed similar numbers of B and T cells (top, using TC⁺ HSC alone were all IgM^b, others were predominantly IgM^a). Increase in B220^loCD5⁺ B cells of IgM^b B1a and CLL development are marked. PerC, peritoneal cavity. Representative of six transfer cases using 2–3-mo-old mice. TC⁺ mice together with TC^– mice. (B) Cotransfer of purified B1 B cells from ≤2-mo-old TC⁺C.B17 mice with TC^– BALB/c BM into C.B17.scid mice, monitoring PBL after transfer. (C) Transfer of B1a cells from 10-d-old TC^+/− mice and development of CLL. 10⁶ purified sB1a B cells (5 × 10⁵ TC⁺) from 12 10-d-old (TC^+/− x CB17)F₁ mice were transferred together with 5 × 10⁶ TC^– BM cells into CB.17. mice. (left) Surface IgM expression of sB1a cells. (right) Frequency of B1a cells in PBL of recipient after adoptive transfer (middle). Characteristic mouse CLL CD5⁺B220⁺ phenotype and lymphomatous spleen B cells (thick black line) compared with original transferred B1a cells (thin black line and CD5⁻B220⁺ B cells (gray). Expression of hTCL1 Tg in this CLL is visualized by cytoplasmic staining (gray, second step control). Total leukemia B cell number indicated as a sum of IgM^b+ B cells in PBL (2 × 10⁸), spleen (4.6 × 10⁸), and peritoneal cavity (1.8 × 10⁸), as compared with the size of the initial TC⁺ inoculum. Representative of 15 cases with CLL/lymphoma generation by ≤2-mo-old mice. TC⁺ B1 B cell transfer. (D) H&E staining of spleen section with CLL/lymphoma of day 10 B1a cell origin. (E) Splenomegaly 8 mo after TC⁺ pB1a transferred together with TC^– BM cells, compared with a recipient of TC^– BM alone. (F) Summary of CLL/lymphoma incidence by purified B1 B cell transfer from ≤2-mo-old TC⁺ mice.