We found that short-term culture medium and homogenate of casein-induced rat peritoneal polymorphonuclear leukocytes (PMN) markedly induced collagenase and prostaglandin E, (PGE,) production by normal rat synovial cells and these effects were abrogated by anti-(rat interleukin-la) (IL-la) polyclonal antibodies. However, collagenase activity and PGE, induced by recombinant rat IL-la were less than those induced by rat PMN culture medium. It was also proved by radioimmunoassay that rat PMN culture medium contains a relatively small amount of IL-la. The introduction of IL-la-deleted PMN culture medium and recombinant rat IL-la together into the synovial cell culture system revealed that IL-ladeleted PMN culture medium has a significant enhancing activity on IL-la-induced synovial cell collagenase and PGE, production. This new factor, which was shown to be a negatively charged protein of about 80 kDa, may have important roles in connective tissue destruction and chronic inflammation in diseases such as rheumatoid arthritis.