Liposomes as immunological adjuvants in eliciting antibodies specific to the synthetic polypeptide poly(LTyr, LGlu)-poly(DLAla)--(LLys) with high frequency of site-associated idiotypic determinants

Abstract

The antibody response to the synthetic polypeptide, poly(LTyr, LGlu)-poly(DLAla)--poly(LLys), [(T,G)-A--L], injected entrapped in liposomes which served as adjuvant has been analyzed. The liposomes used were composed of phosphatidylcholine, cholesterol, dicetylphosphate and DL alpha-tocopherol (molar ratios as 4:3:0.1:0.5) and therefore, were negatively charged. Since the (T,G)-A--L is also negatively charged, no free complexes were formed. The (T,G)-A--L was found to be entrapped inside the enclosed volume of the liposomes, and no (T,G)-A--L antigenic determinants could be detected on the liposomal membranes. Injection of high-responder C3H.SW (H-2b) mice with (T,G)-A--L-bearing liposomes demonstrated that the i.p. and the i.v. routes of immunization were efficient in eliciting (T, G)-A--L specific antibodies, whereas the i.d. injection led to poor antibody responses. The latter route of immunization is the most effective when (T,G)-A--L is injected in complete Freund's adjuvant (CFA). When low doses (0.1 and 1 microgram) of (T, G)-A--L were used for immunization, the liposomes were better adjuvants than CFA. The effectiveness of the liposomes as immunological adjuvants was also shown in their ability to induce high-potential, primed memory cells. The pattern of low (H-2k,a) and high (H-2b) responsiveness to (T,G)-A--L was retained following immunization with (T,G)-A--L entrapped in liposomes, as tested in two pairs of congenic strains. (T,G)-A--L-specific antibodies induced by injection with 1 microgram antigen entrapped in liposomes bear the (T,G)-A--L site-related idiotypic markers of C3H.SW (Igh-1a) mice in a significantly higher frequency than the homologous idiotypes, namely the antibodies elicited in this strain against (T,G)-A--L in CFA. Thus, liposomes may serve as adjuvants for the production of relatively restricted (T,G)-A--L-specific antibodies of high quality.