Fig. 5.
BM-derived DCs expanded from huAIRE-Tg in vitro reflect the developmental signature of DCs in vivo and diabetogenic pathology. (A) Three populations including pDCs (B220+CD11blow), CD11b+ DCs (B220−CD11b+CD24−) and Xcr1+ DCs (B220−CD11b+CD24+) were induced by treatment with BM cells from non-Tg, heterozygous 2m9L-Tg and heterozygous 1m4L-Tg with Flt3L. Expression of Xcr1 from each population (pDC, green line; CD11b+ DC, blue line; Xcr1+ DC, red line) is shown on the right. (B) A summary of the results in (A) is shown. The percentages (upper) and absolute numbers of Xcr1+ DCs (lower) were reduced in heterozygous 2m9L-Tg and, to a lesser extent, in heterozygous 1m4L-Tg. **P < 0.01; *P < 0.05. (C) Expression of huAIRE from in vitro-expanded DC populations was evaluated by flow cytometric analysis. One representative experiment from a total of two repeats is shown. (D) All the heterozygous 1m4L-Tg (n = 10) but one showed pathological changes in β-islets, whereas homozygous 1m4L-Tg showed significantly milder pathological changes (n = 8) compared with those from heterozygous 1m4L-Tg. Pathological changes in the pancreas were evaluated with H&E staining without being informed of the detailed condition of the individual mice. White column, no pathological change; half-filled column, peri-insulitis and/or peri-vasculitis; full-filled column, massive insulitis. M, male: F, female. All the mice were 7.5–8 months old. Mice shown in (E) are marked with asterisks. (E) Two representative examples (one female and one male) of histology from heterozygous and homozygous 1m4L-Tg summarized in (D) are shown. Scale bar: 100 µm.