Abstract

DNA immunization is a potentially important approach to vaccination. It has been shown to work in many animal models, producing immune responses which can counter viruses, bacteria, and tumors. However the mechanisms which underpin this approach remain poorly defined. This proposal is aimed towards analyzing these mechanisms, and using the accrued knowledge to manipulate and optimize DNA vaccines. The proposal has four specific aims.
Aim 1. To identify the cells which take up and express DNA following intramuscular injection, and which present antigen to T cells. It is known that DNA is expressed in muscle cells; are APCs also transfected? We shall use cloned CTL as probes to identify the cells actually presenting antigen following DNA immunization.
Aim 2. To precisely identify which cells induce immunity, and to determine whether muscle cells are important. That cells can be recognized by T cells does not imply that they can induce responses. Can we use cell sorting & transfer to identify the cells responsible for induction of immunity? Aim 3. To evaluate the role of antigen release in DNA immunization, and to identify the underlying mechanisms. Does induction of antibody & CD4+ T cells require antigen release into the humoral phase? If so, what mechanisms underlie this release? Does T cell mediated lysis play a role, and if so, what are the roles of the perforin & fas pathways? Does T cell lysis subsequently limit the immune response? Aim 4. To use the accumulated knowledge to optimize DNA immunization. The knowledge from aims 1-3 will be drawn together to optimize induction of antibodies, CD4+ T cells, and CD8+ T cells each of which probably will have unique requirements for optimal responses.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI037186-05
Application #
2886975
Study Section
Experimental Virology Study Section (EVR)
Program Officer
Ash-Shaheed, Belinda
Project Start
1994-09-30
Project End
2003-03-31
Budget Start
1999-04-01
Budget End
2000-03-31
Support Year
5
Fiscal Year
1999
Total Cost
Indirect Cost

  Institution

Name
Scripps Research Institute
Department
Type
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92037

  Publications

Benning, Nicola; Hassett, Daniel E (2004) Vaccinia virus infection during murine pregnancy: a new pathogenesis model for vaccinia fetalis. J Virol 78:3133-9
Leifert, Jens A; Rodriguez-Carreno, Maria P; Rodriguez, Fernando et al. (2004) Targeting plasmid-encoded proteins to the antigen presentation pathways. Immunol Rev 199:40-53
Leifert, J A; Holler, P D; Harkins, S et al. (2003) The cationic region from HIV tat enhances the cell-surface expression of epitope/MHC class I complexes. Gene Ther 10:2067-73
Leifert, Jens A; Whitton, J Lindsay (2003) ""Translocatory proteins"" and ""protein transduction domains"": a critical analysis of their biological effects and the underlying mechanisms. Mol Ther 8:13-20
Leifert, J A; Harkins, S; Whitton, J L (2002) Full-length proteins attached to the HIV tat protein transduction domain are neither transduced between cells, nor exhibit enhanced immunogenicity. Gene Ther 9:1422-8
Zhang, Jie; Silvestri, Nicole; Whitton, J Lindsay et al. (2002) Neonates mount robust and protective adult-like CD8(+)-T-cell responses to DNA vaccines. J Virol 76:11911-9
Whitton, J L (2002) Designing arenaviral vaccines. Curr Top Microbiol Immunol 263:221-38
Theil, D J; Tsunoda, I; Rodriguez, F et al. (2001) Viruses can silently prime for and trigger central nervous system autoimmune disease. J Neurovirol 7:220-7
Whitton, J L; Fujinami, R S (2001) DNA immunization and central nervous system viral infection. Adv Virus Res 56:243-73
Leifert, J A; Lindencrona, J A; Charo, J et al. (2001) Enhancing T cell activation and antiviral protection by introducing the HIV-1 protein transduction domain into a DNA vaccine. Hum Gene Ther 12:1881-92

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