Tuftsin-enhanced thymidine incorporation by murine splenic monocytes

Tuftsin (Thr-Lys-Pro-Arg), a natural immunomodulating peptide originally found to stimulate phagocytosis by polymorphonuclear leukocytes (PMN), is now known to bind to both PMN and monocyte-macrophages, affecting many of their functions. Administration of tuftsin induces leukocytosis in vivo. We have recently observed that while tuftsin remains in the cytoplasm upon binding and internalization in human PMNs, it translocates into the human monocyte nucleus, suggesting that tuftsin may directly affect growth of monocytes. We have therefore examined the effect of tuftsin on [3H]thymidine incorporation in fractions of murine splenocytes to identify a cell population responding to tuftsin. Tuftsin showed the greatest effect in [3H]thymidine incorporation of splenocytes over controls at optimum conditions of 2% fetal bovine serum and 1 microgram/ml of tuftsin. Splenocyte fractionation by Lymphocyte Separation Medium indicated that tuftsin primarily affects the mononuclear cell fraction; further fractionation revealed that tuftsin affects mostly the monocytes that adhered to plastic. We subsequently further purified the splenic monocytes by repeated plastic adhesion and Percoll gradient separation, to show that tuftsin increases [3H]thymidine incorporation of these highly purified monocytes.