HLA (Class I) antigens are ubiquitous in their cellular distribution and, while their function in major histocompatibility complex (MHC)-restricted phenomena are clear, their function on other cells, such as platelets, is not so obvious. We now report that several anti-HLA monoclonal antibodies (including an anti-beta 2 microglobulin antibody) selectively affect platelet function in that three different anti-HLA monoclonal antibodies caused not only the aggregation of human platelets, but also caused the release of 14C-serotonin. In addition, the anti-HLA antibodies could selectively block the binding of several platelet agonists such as collagen, adrenalin, ADP, but not the binding of others such as thrombin and arachidonic acid. In blocking studies there also appeared to be an association between platelet glycoprotein IIb-IIIa and HLA Class I antigens. We propose that both heavy and light chains of Class I HLA antigens on platelets may be involved in platelet aggregation and release and suggest an additional role for HLA antigens on platelets.