Search

This paper describes the effect of altering liposomal membrane composition on humoral and cellular immunogenicity of haptenated liposomes in mice. Antibody formation was determined by enumeration of direct, plaque-forming cells in the spleen and delayed-type hypersensitivity (DH) was measured with a footpad swelling test. Humoral immunogenicity of haptenated liposomes was strongly influenced by membrane phospholipid, cholesterol and charged amphiphile composition. Haptenated liposomes prepared from phospholipids with a low (diolcoyl- and dilauroyl-phosphatidylcho- line) or high (distearoyl phosphatidylcholine) phase- transition temperature were less immunogenic than those prepared from phospholipids with an intermediate phase-transition temperature (dipalmitoyl phosphatidylcholine and sphingomyelin). In general, increasing the amount of liposomal membrane cholesterol induced a higher humoral response. These results are discussed in relation to liposomal membrane fluidity. Induction of an optimal DH with haptenated liposomes did not occur in the absence of the adjuvant dimethyl dioctadecyl ammonium bromide (DDA). When DDA was used, alterations in membrane composition did not influence cellular immunogenicity. From these results it was concluded that ‘intermediate’ liposomal membrane fluidity is the most important requirement for induction of optimal antibody formation with haptenated liposomes and that a certain physicochemical configuration of the antigen, provided by the adjuvant DDA, is a prerequisite for induction of DH. [ABSTRACT FROM AUTHOR]

This paper describes the induction of delayed-type hypersensitivity (DH) in the mouse and guinea-pig to haptenated liposomes. The tripeptide- enlarged hapten 3-(p-azobenzenearsonate)-N-acetyl- L-tyrosylglycylglycine (A) was coupled to phosphatidylethanolamine (PE) and incorporated into liposomal membranes (A PE-liposomes). In mice DH was measured as footpad swelling and in guinea pigs by skin testing. To induce hapten A-specific DH in mice with A-PE- liposomes the application of the cationic, surface- active lipid, dimethyl dioctadecyl ammonium bromide (DDA) was necessary. The use of Freund's complete adjuvant (FCA) did not result in the induction of DH to hapten A. In guinea-pigs, however, FCA and DDA had equally good adjuvant properties in the induction of DH. The time course of the DH and the optimal time interval between immunization and elicitation were determined for the mouse system. Also, the effect of dose and epitope density was studied in that system. Cyclophosphamide treatment, before immunizing mice with A-PE-liposomes and DDA, resulted in greatly impaired DH, probably caused by the short lifetime of the integrity of liposomes after intracutaneous administration to mice. The results make it very likely that presentation of hapten A in a liposomal or micellar structure is required to induce a cellular immune response to this hapten in mice. [ABSTRACT FROM AUTHOR]

Delayed-type hypersensitivity (DH) and acquired cellular resistance (ARC) to Listeria monocytogenes in mice was studied following immunization with killed bacteria in combination with Freund's complete adjuvant or the adjuvant dimethyldioctadecylammonium bromide (DDA). Intracutaneous or intraperitoneal injections of killed listeria mixed with Freund's complete adjuvant did neither result in DH nor in ACR. Intracutaneous injections of killed listeria and DDA resulted in an antigen-dose dependent DH but not in ACR. Intraperitoneal injection of listeria and DDA, however, induced ACR but no DH. Optimal conditions for the induction of ACR were simultaneous intraperitoneal injection of 15 mg DDA/kg body weight and 107 or 108 listeria. The optimal interval between immunization and challenge was 7 days. No protection was found against challenge with a lethal dose of Salmonella enteritidis, suggesting that the protection is specific. Intraperitoneal injection of mice with DDA resulted in inhibition of phagosomelysosome fusion in macrophages harvested 24 h later. Interference with macrophage activity is discussed as one of the possible mechanisms for the adjuvant effect of DDA. [ABSTRACT FROM AUTHOR]

Immunization of mice with antigen mixed with the cationic surface active lipid dimethyl dioctadccyl ammonium bromide (DDA) produced delayed type hypersensitivity (DH), measured as a footpad swelling. The DH to sheep red blood cells or dinitrophenyl conjugated with bovine serum albumin (DNP28-BSA) in DDA exceeded the response or the same antigens in Freund's Complete Adjuvant (FCA) significantly. Treatment of mice with CY 8 hr prior to the injection of antigen in FCA or DDA resulted in delay of the onset of footpad swelling past day 5 and in elimination of the differences in the response due to the adjuvants. Immunization with carrier or hapten-carrier complexes with different epitope density in DDA and elicitation with the homologous and heterologous antigens revealed that the DH was DNP-specific. In viva priming with DNP28-BSA in DDA and in vitro stimulation with the same antigen resulted in peak responses which were twice as high and were reached almost twice as fast as the earlier found response following immunization in FCA. The advantages of DDA as adjuvant over covalently linked fatty acid chains and over FCA are discussed. [ABSTRACT FROM AUTHOR]

The cell types involved in the cellular immune response were studied with the GVH in vitro as a test system. Comparison of the activities of cells of different lymphoid organs in the GVH in vitro showed the cells of peripheral lymph glands and thoracic duct to be the most active. Second in activity were the cells from peritoneal exudate, mesenterial lymph nodes and spleen. Weak activity was found in thymus and bone marrow cells, whereas cells from Peyer's patches showed no activity at all. Treatment of donor animals with ATS, cortisone or cyclophosphamide simultaneously with (acceptor-) transplantation antigen, resulted in spleen cells which were less active in the GVH in vitro. No such results were obtained after treatment of donor animals with dimethylbenzanthracene or cyclo- phosphamide without antigen. Fractionation of donor spleen cells on a bovine serum albumin density gradient resulted in an accumulation of activity in one of the high density fractions (D), whereas this was reduced in the low density fractions (C, B and A) and absent in the highest density fraction (E). Separation of thymus cells on a glass bead column coated with transplantation antigen resulted in augmented activity of the adherent cell fractions, while the effluent cells were less active than the original cells. Fractionation of cells on an uncoated column resulted in adherent and non-adherent cells which were both inactive in the GVH in vitro. [ABSTRACT FROM AUTHOR]