Figure 4.

Long-term treatment with pyridoxamine impairs neutrophil adhesive function in venules of tumor necrosis factor (TNF)-α-challenged sickle cell disease (SCD) mice. Male (A-P and T-U) and female (Q-U) SCD mice were given acidic water or pyridoxamine (2 g/L) in the drinking water, starting at 2 weeks after bone marrow transplantation and continued for 5.5 months. Intravital microscopy with TNF-α-challenged SCD mice was performed as described in Figure 2. (A) Timeline for the treatment with acidic or pyridoxamine-containing water, TNF-α injection, imaging, and recording survival times. (B) Rolling influx of neutrophils. (C) Cumulative frequency of the rolling velocity of neutrophils. (D) Number of adherent neutrophils. (E) Integrated median fluorescence intensity values of anti-CD42c antibodies (F platelets) were plotted over time. (F) Survival curves of SCD mice during or after intravital microscopy. Data represent the mean±standard deviation (SD) (n=58-62 venules in 7-8 mice per group). (G-I) Kidney, spleen, and liver were obtained from unchallenged or TNF-α-challenged SCD mice treated with acidic water or pyridoxamine- containing water after recording the survival time. (G) Representative hematoxylin and eosin staining. Bar=50 µm. (H) Glomerular size. (I) Number of adherent leukocytes on the hepatic vessel wall. Data represent the mean±SD (n=7 mice per group). ^**P<0.01, ^***P<0.001, and ^****P<0.0001, Student’s t-test (B, D, H, and I), Mann-Whitney U test (C), and Mantel-Cox log-rank test (F).