Differential effects of liposome-incorporation on liver macrophage activating potencies of rough lipopolysaccharide, lipid A, and muramyl dipeptide. Differences in susceptibility to lysosomal enzymes.
We investigated the in vitro activation of rat liver macrophages to a tumor-cytotoxic state with muramyl dipeptide (MDP), rough LPS (Re-LPS) and lipid A in both a free and liposome-encapsulated form. The tumor cytotoxic state of the liver macrophages was determined with a [methyl-3H]thymidine release assay using C26 colon adenocarcinoma cells as target cells. As was shown previously, the encapsulation of MDP within multi-lamellar phospholipid vesicles greatly enhanced the activating potency of the drug; by contrast, encapsulation of Re-LPS or lipid A significantly reduced the activation of macrophages as compared to the free form of these agents. At a dose of 1 ng of free Re-LPS per ml a significant induction of tumor cell lysis was observed whereas a maximal level was obtained at a concentration of approximately 10 ng/ml. By encapsulation of Re-LPS in liposomes the activating potency diminished 20- to 100-fold. The minimal concentration required to induce detectable macrophage activation with free lipid A was 10 ng/ml, while liposome-encapsulated lipid A did not induce any detectable tumor cell lysis up to a concentration of 200 ng/ml. After a 1-h pre-incubation with a lysosomal fraction from rat liver at pH 4.8, the macrophage-activating potency of Re-LPS and lipid A was diminished by up to 95% whereas MDP remained fully active under these conditions. We conclude that, due to endocytic uptake of liposome-incorporated Re-LPS and lipid A and subsequent intralysosomal degradation, these immunomodulators are inactivated with respect to their potency to activate liver macrophages to tumor cytotoxicity.
Daemen
T
T
Laboratory of Physiological Chemistry, University of Groningen, The Netherlands.
Veninga
A
A
Dijkstra
J
J
Scherphof
G
G
D000119:Acetylmuramyl-Alanyl-Isoglutamine / Q000494:pharmacology; D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000818:Animals; D003602:Cytotoxicity, Immunologic / Q000187:drug effects; D005260:Female; D008050:Lipid A / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008081:Liposomes / Q000276:immunology*; D008099:Liver / Q000276:immunology*; D008247:Lysosomes / Q000201:enzymology*; D008262:Macrophage Activation / Q000187:drug effects*; D008264:Macrophages / Q000201:enzymology / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats; D011919:Rats, Inbred Strains; D014407:Tumor Cells, Cultured / Q000187:drug effects
D000276:Adjuvants, Immunologic; D008050:Lipid A; D008070:Lipopolysaccharides; D008081:Liposomes; D000119:Acetylmuramyl-Alanyl-Isoglutamine