The in vitro functional activity of bovine peripheral blood lymphocytes and the population dynamics of the major peripheral blood leukocyte subpopulations were investigated following the administration of recombinant bovine interferon-alpha I1 (rBoIFN-alpha I1). The intramuscular injection of rBoIFN-alpha I1 induced a 24-h lymphopenia characterized by a decrease in both circulating T and non-T/non-B lymphocytes. An increased CD4/CD8 ratio indicated that there was a relatively greater depletion of the CD8 equivalent subpopulation of T lymphocytes. These changes in lymphocyte trafficking were observed in the absence of a cortisolemia. Coincidental with the lymphopenia, there was a marked decrease in the amplitude of mitogen-induced lymphocyte proliferation responses that probably was caused by a numerical deficit rather than functional deficit in the responder T cells.