A novel method to quantify the turnover and release of monocytes from the bone marrow using the thymidine analog...

The present study was designed to develop methods to study the production and release of monocytes from the bone marrow using the thymidine analog 5'-bromo-2'-deoxyuridine (BrdU). Dividing monocytes in bone marrow were labeled with BrdU (MOBrdU), and their release into the blood and disappearance from the circulation were monitored using a double immunostaining method. The first MOBrdU appeared in the circulation 4 h after labeling with BrdU and peaked at 18 h when 34.3 +/- 5.8% of monocytes were labeled. The calculated transit time of monocytes through bone marrow was 38.1 +/- 3.1 h in control rabbits with a half-life (T1/2) of 12.7 h. Instillation of Streptococcus pneumoniae into the lung accelerated the release of monocytes from bone marrow (peak at 10 h) and shortened their bone marrow transit time (27.1 +/- 1.8 vs. 22.6 +/- 0.6, vehicle vs. pneumonia; P < 0.05). We conclude that this nonradioisotope method provides a novel way to monitor monocyte kinetics and confirmed previous reports that a focal pneumonia shortens monocyte marrow transit and increases their release into the circulation.