Polymorphonuclear leukocytes (PMN) contribute to post-ischemic injury in many organs and in a variety of clinical situations. PMN accumulate in both lungs during unilateral lung ischemia in sheep, but the mechanism has not been defined. In this study, we tested the hypothesis that PMN accumulation is a response to chemotactic signals generated during lung ischemia. Chemotactic activity was measured in a modified Boyden chamber using normal sheep PMN as the responding cells. Increased chemotactic activity was observed in both plasma and lung lymph in a time-dependent manner after ischemia. These data indicate that a chemotactic substance immunoreactive to interleukin-8 antibody is formed as a result of unilateral lung ischemia in sheep in vivo and is a possible mediator of PMN inflammation in this model.