Fig. 3.
Rig-I deficiency-associated granulopoiesis is ascribed to an autonomous defect of hematopoietic cells. (A) Total of 1 × 106 pooled BM cells from Rig-I+/+ or Rig-I−/− mice at 6 months of age (n = 3) were transplanted into the lethally irradiated syngeneic mice (n = 7). The regeneration of BM or PBL granulocytes was measured 7 weeks later. The color images on the right show May-Grünwald-Giemsa–stained cytospins of PBLs from primary recipients. The DNA-PCR analysis of PBLs showed that >90% of leukocytes were derived from donors (data not shown). (B) Bone marrow cells from the primary recipients were harvested and further transplanted into the secondary recipients. A representative sample of flow cytometric analysis of granulocytes in PBLs, BM, and spleen was performed 7 weeks later (n = 6). (C) Primary colony-forming potentials of Rig-I+/+ or Rig-I−/− BM myeloid progenitors responded to the stimulations by different cytokines as indicated (mean ± SD) (GM-CSF colony, P = 0.02). (D) Percentages of c-Kit+Gr-1lo granulocytic progenitors (GPs) within the retrieved colony cells. (E) May-Giemsa staining of macrophages (black arrowheads), granulocytes (white arrowheads) and immature myeloid cells (gray arrowheads) were generated from primary Rig-I+/+ and Rig-I−/− BM myeloid progenitors stimulated by GM-CSF.