DNA immunization with a herpes simplex virus 2 bacterial artificial chromosome
Clement A Meseda1, Falko Schmeisser, Robin Pedersen
1Laboratory of DNA Viruses, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, USA.
Virology
|February 20, 2004
Summary
A novel herpes simplex virus 2 (HSV-2) bacterial artificial chromosome (BAC) was developed. This HSV-2 BAC DNA induced protective immunity in mice, showing potential for new HSV-2 vaccines.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Herpes simplex virus 2 (HSV-2) poses significant public health challenges.
- Development of effective HSV-2 vaccines remains a critical unmet need.
- Bacterial artificial chromosome (BAC) technology offers a platform for genetic manipulation of viral genomes.
Purpose of the Study:
- To construct and characterize a herpes simplex virus 2 (HSV-2) bacterial artificial chromosome (BAC).
- To evaluate the immunogenicity and protective efficacy of the HSV2-BAC DNA in a mouse model.
- To demonstrate the utility of BAC technology for creating novel HSV-2 vaccine candidates.
Main Methods:
- HSV-2 BAC construction via homologous recombination into the thymidine kinase gene.
- Transformation of recombinant viral DNA into E. coli for BAC isolation and genotyping.
- Transfection of HSV2-BAC DNA into mammalian cells to assess infectivity and phenotype.
- Immunization of mice with HSV2-BAC DNA and subsequent challenge with live HSV-2.
- Construction and evaluation of a gD-deleted HSV2-BAC mutant.
Main Results:
- Successfully constructed and isolated infectious HSV2-BAC DNA capable of generating thymidine kinase-negative virus.
- HSV2-BAC DNA immunization elicited strong HSV-2 specific antibody responses, comparable to live virus.
- HSV2-BAC immunization provided significant protection against lethal HSV-2 challenge.
- A recombinant HSV2-BAC with a deleted glycoprotein D (gD) gene was constructed and shown to be replication-deficient without complementation.
- Immunization with the gD-deleted HSV2-BAC also induced protective immunity.
Conclusions:
- HSV-2 BAC technology is feasible for developing both replicating and non-replicating HSV-2 vaccine candidates.
- BAC technology facilitates the construction and maintenance of novel HSV-2 vaccines.
- This approach offers a powerful tool for dissecting the immune response to HSV-2 and developing new vaccines.
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