Zusammenfassung
We have analysed the influence of size, intracellular localisation, and sorting of various human immunodeficiency virus type 1 (HIV-1)-derived Gag and Env polypeptides containing well defined H2(d)-restricted cytotoxic T lymphocyte (CTL) epitopes on the induction of a Immoral and cellular immune response after DNA vaccination. Thus, expression vectors were generated based on RNA- and ...
Zusammenfassung
We have analysed the influence of size, intracellular localisation, and sorting of various human immunodeficiency virus type 1 (HIV-1)-derived Gag and Env polypeptides containing well defined H2(d)-restricted cytotoxic T lymphocyte (CTL) epitopes on the induction of a Immoral and cellular immune response after DNA vaccination. Thus, expression vectors were generated based on RNA- and codon-optimised genes encoding (i) budding competent full-length Gag, (ii) a myristylation defect mutant GagMyr(-), (iii) the isolated p24 capsid moiety of Gag as well as variants of these proteins, which were C-terminally fused HIV gp120-derived V3 epitope (R10I), respectively. These constructs were compared to different minitopes each encoding one of the H2(d)-restricted Gag epitopes A9I and E10F or the V3 epitope R10I that were directly linked to the C-terminus of an Ad2-E3 protein-derived ER signal peptide. Immunological evaluation of these constructs in BALB/c mice revealed that both, the budding competent as well as the intracellular Gag proteins were-irrespective of their molecular weights-equally efficient in the priming of Gag-specific humoral and cellular immune responses. In addition, the capacity of these constructs to stimulate Gag-specific humoral as well as H2-K-d and H2-L-d restricted cellular immune responses was not influenced by C-terminal fusion of the immunodominant H2-D-d restricted V3 epitope. Chimeric GagV3 polyproteins encoding all three major CTL epitopes within a continuous polyprotein were more efficient to stimulate epitope-specific cellular immune responses than the selected minitopes. In addition, the minitopes failed to induce epitope-specific antibody responses. These results clearly show the advantages of complex polypeptides over minitopes regarding the induction of strong humoral and cellular immune responses. (C) 2004 Elsevier Ltd. All rights reserved.
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