Directed locomotion of mouse peritoneal neutrophils under agarose was studied, and activity of hydrogen peroxide (H2O2) as a chemoattractant was tested in its concentration range of 10(-6) to 10(-3) M. It has been found that H2O2 at low concentrations (about 10 microM) induces chemotactic activity. This activity was not affected by the presence of serum in the agarose medium. Use of bovine serum albumin instead of the heat-inactivated bovine serum in the medium had no effect on cell locomotion. The H2O2-induced chemotaxis was significantly reduced by catalase. Involvement of [Ca2+]i transients in the H2O2-induced chemotactic response was shown. These data indicate that H2O2 itself in small quantities can act as a chemoattractant without interacting with a plasma precursor to form a chemotactic factor. It has been suggested that H2O2 may form an important link similar to the second messenger in communication between the cells.