The C-band heterochromatin polymorphisms of chromosomes 1, 9, and 16 were studied on peripheral lymphocytes of 67 children with acute lymphoblastic leukemia (ALL) and 50 control individuals. A statistically significant difference between patients and controls was found for large heterochromatin regions (level 3) of chromosomes 1 and 9 (P < 0.001) and for small heterochromatin regions (level 1) of chromosome 16 (P < 0.001). The patients also showed a significant increase in chromosomes 1 and 9 heteromorphism with respect to controls (P < 0.001).