Prophylactic potential of liposomized integral membrane protein of Plasmodium yoelii nigeriensis against blood stage infection in BALB/c mice.
ABSTRACT Triton X-114 phase separated integral membrane proteins (IMPs) of a multidrug resistant strain of Plasmodium yoelii nigeriensis (P. yoelii) were screened for their potential to impart protection against malaria infection in BALB/c mice. As revealed by immunoblotting, antibodies present in parasite specific sera from convalescent (protected) as well as immunized (partially protected) animals recognized different membrane proteins. A thorough investigation reveals that P. yoelii specific convalescent sera recognized IMPs with molecular masses ranging from 21 to 81 kDa. Among various membrane proteins, the IMPs corresponding to 81 and 66 kDa molecular weight were highly prominent in the immunoblots probed with the sera from convalescent animals, whereas sera from immunized animals failed to produce impressive band pattern. Immunofluorescence assay revealed that the 66-kDa IMP specific antibodies reacted with fixed smears of mature schizonts and merozoites. Further immunization with 66 kDa IMP (PyIMP) purified through polyclonal IgG sepharose 4B affinity did not impart effective immune response (in its free form) and could provided partial protection only. On the other hand, animals immunized with 66 kDa PyIMP entrapped in phosphatidyl-choline/cholesterol (PC/chol) liposomes protected BALB/c mice against lethal P. yoelii challenge.
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ABSTRACT: Apical membrane antigen 1 (AMA1) is considered one of the leading candidates for inclusion in a vaccine against blood stages of Plasmodium falciparum. Although the ama1 gene is relatively conserved compared to those for some other potential vaccine components, numerous point mutations have resulted in amino acid substitutions at many sites in the polypeptide. The polymorphisms in AMA1 have been attributed to the diversifying selection pressure of the protective immune responses. It was therefore of interest to investigate the impact of sequence diversity in P. falciparum AMA1 on the ability of anti-AMA1 antibodies to inhibit the invasion of erythrocytes in vitro by P. falciparum merozoites. For these studies, we used antibodies to recombinant P. falciparum 3D7 AMA1 ectodomain, which was prepared for testing in early clinical trials. Antibodies were raised in rabbits to the antigen formulated in Montanide ISA720, and human antibodies to AMA1 were isolated by affinity purification from the plasma of adults living in regions of Papua New Guinea where malaria is endemic. Both rabbit and human anti-AMA1 antibodies were found to be strongly inhibitory to the invasion of erythrocytes by merozoites from both the homologous and two heterologous lines of P. falciparum. The inhibitory antibodies targeted both conserved and strain-specific epitopes within the ectodomain of AMA1; however, it appears that the majority of these antibodies reacted with strain-specific epitopes in domain I, the N-terminal disulfide-bonded domain, which is the most polymorphic region of AMA1.Infection and Immunity 06/2001; 69(5):3286-94. · 4.07 Impact Factor
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ABSTRACT: In previous study, we demonstrated the potential of Escherichia coli (E. coli) lipid liposomes (escheriosomes) to undergo membrane-membrane fusion with cytoplasmic membrane of the target cells including professional antigen presenting cells. Our present study demonstrates that antigen encapsulated in escheriosomes could be successfully delivered simultaneously to the cytosolic as well as endosomal processing pathways of antigen presenting cells, leading to the generation of both CD4(+) T-helper and CD8(+) cytotoxic T cell response. In contrast, encapsulation of same antigen in egg phosphatidyl-choline (egg PC) liposomes, just like antigen-incomplete Freund's adjuvant (IFA) complex, has inefficient access to the cytosolic pathway of MHC I-dependent antigen presentation and failed to generate antigen-specific CD8(+) cytotoxic T cell response. However, both egg PC liposomes as well as escheriosomes-encapsulated antigen elicited strong humoral immune response in immunized animals but antibody titre was significantly higher in the group of animals immunized with escheriosomes-encapsulated antigen. These results imply usage of liposome-based adjuvant as potential candidate vaccine capable of eliciting both cell-mediated as well as humoral immune responses. Furthermore, antigen entrapped in escheriosomes stimulates antigen-specific CD4(+) T cell proliferation and also enhances the level of IL-2, IFN-gamma and IL-4 in the immunized animals.Vaccine 07/2003; 21(19-20):2383-93. · 3.49 Impact Factor
- Journal of Biological Chemistry - J BIOL CHEM. 01/1996; 271(46):29446-29452.