Delivery of cytokines by liposomes: Hematopoietic and immunomodulatory activity of interleukin-2 encapsulated in conventional liposomes and in long- circulating liposomes

Although liposomal delivery of interleukin-2 (IL-2) and other cytokines improves their pharmacokinetics and biologic activity in vivo, there are no comparative functional studies of various liposomal formulations as cytokine carriers. In the present investigation, recombinant human IL-2 was encapsulated in two formulations of large (mean diameter 0.75-1.5 μm) multilamellar vesicles (MLV, referred to as conventional liposomes) or in small (mean diameter, 60 nm), unilamellar, long-circulating liposomes (referred to as sterically stabilized liposomes, SSL). The biologic activity of the liposomal formulations and of free IL-2 was tested in parallel in vitro and in mice. The main observations were as follows: (a) All the liposomal IL-2 (Lip-IL-2) formulations were more efficient than soluble IL-2 in stimulating spleen cell proliferation and lymphokine-activated killer (LAK) cell activation in vitro, particularly at low cytokine doses (1-100 CU/mL). (b) After i.v. injection, the circulation time of MLV-IL-2 and SSL- IL-2 was 7 and 17 times greater, respectively, than that of soluble IL-2. (c) In comparison with IL-2, all Lip-IL-2 formulations caused a marked increase in the leukocyte levels in blood, spleen, and peritoneal exudate, especially in those of myeloid origin (neutrophils, eosinophils, immature granulocytes, and macrophages). (d) Although SSL-IL-2 exhibited the longest circulation time, MLV-IL-2 was more potent in elevating leukocyte levels and in triggering LAK cell activity in vivo. (e) The route of Lip-IL-2 administration greatly affected the immunomodulatory activity in the various compartments. (f) MLV-IL-2 proved to be a much more efficient immunoadjuvant than free IL-2 for influenza subunit vaccines as well as for tumor cell vaccines. These findings lend support to our previous studies in which we demonstrated the superior immunomodulatory activity of liposomal IL-2, and suggest that cytokine pharmacokinetics, biodistribution, and pharmacodynamics are markedly influenced both by liposomal formulation and route of administration.