Figure 3.

Deficiency of Tnfr accelerates bone marrow failure (BMF) and enhances Th1 cell responses in Tak1^mut mice. (A) Survival of Tak1^mutTnfr^−/− and Tak1^mut and Tnfr^−/− mice were recorded and compared (*P<0.05, compared to Tnfr^−/−mice; #P<0.05 compared to Tak1^mut mice). Peripheral blood (PB) and bone marrow (BM) were collected from 4-month old Tak1^mutTnfr^−/− mice and age-matched Tak1^mut and Tnfr^−/− mice. (B) White blood cells, (WBC), red blood cells (RBC), hemoglobin (Hb) and platelets (plt) were analyzed using the Hemavet 950 Hematology System. (C) H&E stained bone marrow section (tibia) after decalcification. (D) Number of total nucleated cells (TNCs) in BM from two hind limbs were counted and compared. (E) Representative flow cytometric plots for analysis of BM hematopoietic stem cells (HSCs) and hematopoietic progenitor cells (HPCs). BM cells were first gated on the Lin⁻ population and then analyzed for LK, LSK and LS populations. (F) Percentages of LK, LSK and LS populations in total nucleated cells (TNCs) from BM. (G) Flow analysis of BM CD3⁺ T cells and B220⁺ B cells (gated on lymphocytes). (H) Flow cytometric analysis of naïve T cells (CD62L⁺ CD44⁻) and activated T cells (CD62L^lowCD44^hi) in BM (gated on CD3⁺ cells). (I) Percentages of Th1, Tnfα⁺, Th17, Th2, and Treg cells in BM (gated on CD4⁺ T cells). Ifnγ and Tnfα levels in BM CD4⁺ (J) and CD8⁺ (I) T cells were analyzed by mean fluorescence intensity (MFI) of intracellular antibody staining. (K) Percentages of Tc1, Tnfα⁺, Tc17, Tc2, and Treg cells in BM (gated on CD8⁺ T cells). Data are presented as means±SD. *P<0.05 compared to Tnfr^−/−; ^#P<0.05 compared to Tak1^mut.