title: Heat shock protein micro-encapsulation as a double tool for the improvement of new generation vaccines.
abstract: The modern vaccinology encompasses the recombinant DNA technology, protein and carbohydrate chemistry to obtain safe molecularly defined vaccines. Nevertheless most of the vaccines are poorly immunogenic because a large number of antigens are membrane proteins and consequently they are not present in their active conformation in the vaccine. Others are not as potent because they contain only B epitopes and therefore, cannot stimulate cellular memory. We have been studying the characteristics of the recombinant heat shock protein 18kDa-hsp from Mycobacterium leprae as an alternative carrier protein with a T epitope source to enhance the activity of these second generation vaccines. Here we proved that the 18kDa-hsp acted as carrier, without masking the activity of the carried antigen, with similar immune stimulatory effect when compared with ODN1668. Supramolecular aggregates of 18kDa-hsp and Mice serum albumin (MSA) were obtained using glutaraldehyde as cross linker. The Neisseria meningitides serogroup C polysaccharide (PSC, a B epitope) and the carrier protein 18kDa-hsp were co-encapsulated within Soybean phosphatidylcholine liposomes (SPC: Cho : alpha-Toc, 22 : 5 : 0.18 molar ratio, respectively). These liposomes were prepared in MPB buffer (20 mM phosphate, 295 mM mannitol pH 7.2) in the presence or absence of the ODN1668, TCCATGACGTTCCTGATGCT. When mice were injected with 18kDa-hsp-MSA no antibody against the MSA was observed. This means that the 18kDa-hsp acted as carrier, without masking the carried protein immune activity. Stable liposomes of 150 nm were obtained using mannitol as a cryoprotector. Genetically selected mice when injected with liposomes containing PSC and 18kDa-hsp displayed an antibody titer of 12. In contrast, in those mice injected with free PSC there was no response. The 18kDa-hsp adjuvant effect on the PSC liposomal formulation was comparable to that observed when ODN1668 was co-encapsulated with PSC. Confirming our expectations we observed that the formulation containing 18kDa-hsp conferred a memory response to the carried antigen--the Neisseria meningitidis serogroup C polysaccharide.
authors.affiliation: Lab de Microesferas e Liposomos, C Biotecnologia, Av Vital Brasil, São Paulo, SP. bdacosta@edu.usp.br
authors.forename: Maria Helena
authors.initials: MH
authors.lastname: Bueno Da Costa
authors.forename: Wagner
authors.initials: W
authors.lastname: Quintilio
authors.forename: Martha Massako
authors.initials: MM
authors.lastname: Tanizaki
authors.forename: Osvaldo Augusto
authors.initials: OA
authors.lastname: Sant'Anna
authors.forename: Reto Albert
authors.initials: RA
authors.lastname: Schwendener
authors.forename: Pedro Soares
authors.initials: PS
authors.lastname: de Araujo
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D002942:Circular Dichroism; D004339:Drug Compounding; D016503:Drug Delivery Systems; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006358:Hot Temperature; D008081:Liposomes / Q000378:metabolism; D008297:Male; D051379:Mice; D009345:Neisseria meningitidis / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013696:Temperature; D013997:Time Factors; D014612:Vaccines / Q000378:metabolism / Q000494:pharmacology*
chemical_list: C067864:18 kDa protein, Mycobacterium leprae; D001426:Bacterial Proteins; D008081:Liposomes; D011134:Polysaccharides; D011994:Recombinant Proteins; D014612:Vaccines