Critical factors for liposome-incorporated tumour-associated antigens to induce protective tumour immunity to SL2 lymphoma cells in mice

Physical and immunogenic properties of re- constituted membranes designed for the presentation of tumour-associated antigens (TAA) to the immune system are described. Proteins and lipids of crude membranes of SL2 routine lymphosarcoma cells were partially solubi- lized with octylglucoside. Reconstituted membranes, con- sisting mainly of unilamellar vesicles with a diameter of 0.03-0.15 gm, were formed by detergent removal and were purified by floatation in a discontinuous sucrose gra- dient to remove non-lipid-bound protein. Subcutaneous immunization of syngeneic mice with reconstituted mem- branes or with purified reconstituted membranes induced protection against an intraperitoneal challenge with 103 viable SL2 cells. Reconstituted membranes were more im- munogenic than crude membranes in immunoprotection experiments when compared on the basis of protein dose. Detergent removal was required to obtain an immunogenic presentation form of SL2 membrane antigens and to avoid toxicity associated with the detergent. Reconstitution of SL2 membranes in the presence of exogenous phos- pholipid slightly increased the fraction of protein that as- sociated with the reconstituted membranes. However, the immunogenicity of the solubilized membrane TAA was not significantly affected by the presence of exogenous phospholipid. The reconstitution procedure described may be useful in identifying membrane factors required for the induction of immune responses against TAA. The versatil- ity of the system may be employed to develop safe alterna- tives for whole-cell vaccines.

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