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Posttranscriptional regulatory elements enhance antigen expression and DNA vaccine efficacy.

ABSTRACT: In higher eukaryotes, introns are usually required for efficient pre-mRNA processing. However, some viruses have alternative approaches involving posttranscriptional regulatory elements (PREs) to enhance intronless heterologous gene expression through enabling stability and 3' end formation, and to facilitate the nucleocytoplasmic export of unspliced mRNAs. In the current study, we compared the human cytomegalovirus (hCMV) immediate/early (IE) intronA, as well as virus-derived PREs-the PRE of Hepatitis B virus (HPRE) and Woodchuck Hepatitis virus (WPRE) on their ability to enhance antigen gene expression in vitro and immune responses induced by DNA vaccination in animal. Among all the constructs, the plasmids carrying the HPRE element showed the highest gene expression level in both in vivo and in vitro models. During immunization of mice with low doses (10 microg) of HIV-1 DNA vaccine, only -intronA/+HPRE and +intronA/+HPRE vaccine constructs induced anti-Gag antibodies, although the -intronA/+WPRE construct also elicited antigen-specific cellular immune responses. In addition, pInHGag (+intronA/+HPRE) at a 10 mug dose could induce higher anti-Gag antibody level than that induced by pGag (-intronA/-HPRE) or pInGag (+intronA/-HPRE) at 40 microg dose (p < 0.05). Our data are useful for the optimization of heterologous expression and immunogenicity of DNA vaccines.

SUBMITTER: Sun J

PROVIDER: S-EPMC2719066 | biostudies-literature | 2009 May

REPOSITORIES: biostudies-literature

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Posttranscriptional regulatory elements enhance antigen expression and DNA vaccine efficacy.

Sun Jing J Li Dingfeng D Hao Yanling Y Zhang Yuwei Y Fan Wenling W Fu Jingjing J Hu Yunzhang Y Liu Yong Y Shao Yiming Y

DNA and cell biology 20090501 5

In higher eukaryotes, introns are usually required for efficient pre-mRNA processing. However, some viruses have alternative approaches involving posttranscriptional regulatory elements (PREs) to enhance intronless heterologous gene expression through enabling stability and 3' end formation, and to facilitate the nucleocytoplasmic export of unspliced mRNAs. In the current study, we compared the human cytomegalovirus (hCMV) immediate/early (IE) intronA, as well as virus-derived PREs-the PRE of He ...[more]

PMID: 19388846

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Project description:Peptide subunit vaccines increase safety by reducing the risk of off-target responses and improving the specificity of the induced adaptive immune response. The immunogenicity of most soluble peptides, however, is often insufficient to produce robust and lasting immunity. Many biomaterials and delivery vehicles have been developed for peptide antigens to improve immune response while maintaining specificity. Peptide nanoclusters (PNC) are a subunit peptide vaccine material that has shown potential to increase immunogenicity of peptide antigens. PNC are comprised only of crosslinked peptide antigen and have been synthesized from several peptide antigens as small as 8 amino acids in length. However, as with many peptide vaccine biomaterials, synthesis requires adding residues to the peptide and/or engaging amino acids within the antigen epitope covalently to form a stable material. The impact of antigen modifications made to enable biomaterial incorporation or formation is rarely investigated, since the goal of most studies is to compare the soluble antigen with biomaterial form of antigen. This study investigates PNC as a platform vaccine biomaterial to evaluate how peptide modification and biomaterial formation with different crosslinking chemistries affect epitope-specific immune cell presentation and activation. Several types of PNC were synthesized by desolvation from the model peptide epitope SIINFEKL, which is derived from the immunogenic protein ovalbumin. SIINFEKL was altered to include extra residues on each end, strategically chosen to enable multiple conjugation chemistry options for incorporation into PNC. Several crosslinking methods were used to control which functional groups were used to stabilize the PNC, as well as the reducibility of the crosslinking. These variations were evaluated for immune responses and biodistribution following in vivo immunization. All modified antigen formulations still induced comparable immune responses when incorporated into PNC compared to unmodified soluble antigen alone. However, some crosslinking methods led to a significant increase in desirable immune responses while others did not, suggesting that not all PNC were processed the same. These results help guide future peptide vaccine biomaterial design, including PNC and a wide variety of conjugated and self-assembled peptide antigen materials, to maximize and tune the desired immune response.

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Posttranscriptional regulatory elements enhance antigen expression and DNA vaccine efficacy.

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Posttranscriptional regulatory elements enhance antigen expression and DNA vaccine efficacy.

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