Double labeling and in vitro versus in vivo incorporation of bromodeoxyuridine in patients with acute nonlymphocytic leukemia

Abstract

A monoclonal antibody against bromodeoxyuridine (BrdUrd) was produced, and a rapid slide technique (RPMB technique) was developed for the estimation of S‐phase cells in a population using this antibody. Bone marrow cells from patients with acute nonlymphocytic leukemia (ANIA) were studied by both the RPMB technique and tritiated thymidine (³HdThd) labeling index studies. The percentage of S‐phase cells obtained by each method was compared in 50 samples, and the correlation coefficient was r = 0.89. A “double label” method is also described in which cells were simultaneously incubated with either BrdUrd and ³HdThd or BrdUrd and tritiated cytosine arabinoside (³HAra‐C). The samples were first processed by the RPMB technique and then by autoradiography. Results showed only black grains overlying the nuclei of fluorescent cells in each group. An automated microphotometer was used to quantitate grains and fluorescence from each cell. This demonstrated an almost direct relationship between grains and fluorescence from BrdUrd + ³HdThd slides, whereas different patterns of relationship were noted from BrdUrd + ³HAra‐C slides of leukemic patients. Their implications are discussed in the text. Finally, intravenous infusions of BrdUrd was given to five leukemic patients. S‐phase cells were recognized distinctly within 5 min of starting the infusion. The percentage of Sphase cells was almost identical from in vivo and in vitro samples. Various possibilities of studying the biological behavior of acute leukemias and analyzing cell cycle characteristics are discussed.