Abstract
Human myelogeneous leukemia cells in liquid culture can be induced to mature along the monocyte/macrophage pathway by a maturation inducer derived from the conditioned medium of activated human T lymphocytes. Serum-free conditioned medium was used for the isolation of the T-cell lymphokine. The maturation inducer was purified approximately equal to 6000-fold by ammonium sulfate precipitation, low-salt elution from DEAE-Sepharose CL-6B, gel filtration on Bio-Gel A-0.5m, and NaDodSO4/PAGE under nonreducing conditions. The molecular weight of the maturation inducer was 36,000-58,000 on NaDodSO4/PAGE. Terminal differentiation associated with inhibition of leukemia cell proliferation and expression of mature cell properties was observed with the isolated maturation inducer, identical to the activity observed with the unfractionated conditioned medium. Cell-cycle analysis revealed that the proportion of replicating S-phase cells was reduced from 40% to 7% after initial interaction of the maturation inducer with cells. The differentiating cells simultaneously acquired monocyte antigen, membrane complement receptors, phagocytic function, and monocyte/macrophage morphology. The maturation-inducing activity is dose-dependent, with more inducer causing the development of more mature cells in a shorter time period. The maturation inducer was shown to be stable after pH 2 treatment, independent of interleukin 2 and colony-stimulating factor, devoid of alpha-, beta-, and gamma-interferon, and not affected by antibody to interferon. The maturation inducer may play a role as a physiological regulator of monocytic and leukemia cell development.