engleski

Adjuvant effect of peptidoglycan monomer in liposome based and oil based adjuvant formulations

Peptidoglycan monomer, GlcNAc-MurNAc-L-Ala-D-isoGln-mesoDAP(wNH2)-D-Ala-D-Ala (PGM, PLIVA, Croatia) is a non-toxic, apyrogenic and water-soluble natural compound of bacterial origin. In mice, immunised with ovalbumin (OVA), PGM enhances specific IgG levels (both IgG1 and IgG2a). The goal of this study was to test whether oil based and liposome based adjuvant formulations containing PGM, that we expect to prolong PGM's availability in the organism, would improve its adjuvant efficacy using OVA as an antigen. PGM in Incomplete Freund's adjuvant (IFA) induced significantly higher levels of OVA-specific antibodies (measured by ELISA) in comparison to both PGM and IFA alone. Quantification of OVA-specific IgG1 and IgG2a revealed that Th1/Th2 balance was directed towards Th1 direction with this formulation, similar to the one obtained using Complete Freund's adjuvant (CFA). Liposome based formulations were prepared by encapsulating PGM into large multilamellar negatively charged vesicles using synthetic dipalmitoyl phosphatidylcholine prior to mixing with OVA in saline. Such preparation raised OVA-specific IgG levels in immunised mice sera in comparison to both, PGM in saline, and to empty liposomes. The preliminary analysis of OVA-specific IgG1 and IgG2a subclasses indicated that there was no influence on Th1/Th2 balance. In conclusion, both approaches - liposome based and oil based - improved adjuvant efficacy of PGM. Freund type emulsion with PGM added influenced also Th1/Th2 balance, directing immune response towards Th1 pathway.

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