We have purified five types of lectins, named tachylectins, from circulating hemocytes and hemolymph plasma of the Japanese horseshoe crab, Tachypleus tridentatus. Tachylectin-1 interacts with Gram-negative bacteria probably through 2-keto-3-deoxyoctonate, one of the constituents of lipopolysaccharides (LPS). Tachylectin-1 also binds to polysaccharides such as agarose and dextran with broad specificity. Tachylectin-2 binds to D-GlcNAc or D-GalNAc and recognizes staphylococcal lipoteichoic acids and LPS from several Gram-negative bacteria. In contrast, tachylectins-3 and -4 specifically bind to S-type LPS from several Gram-negative bacteria through a certain sugar moiety on the O-specific polysaccharides (O-antigens). Tachylectin-5 identified in hemolymph plasma has the strongest bacterial agglutinating activity in the five types of tachylectins, and exhibits broad specificity against acetyl group-containing substances. Thus, the innate immune system of horseshoe crab may recognize invading pathogens through a combinatorial method using lectins with different specificities against carbohydrates exposed on pathogens. An encounter of these lectins derived from hemocytes and hemolymph plasma at injured sites, in response to the stimulation of LPS, suggests that they serve synergistically to accomplish an effective host defense against invading microbes and foreign substances. (C) 1999 Elsevier Science Ltd. All rights reserved.