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- Title
Immune responses to T-cell epitopes of SARS CoV-N protein are enhanced by N immunization with a chimera of lysosome-associated membrane protein.
- Authors
Yang, K.; Sun, K.; Srinivasan, K. N.; Salmon, J.; Marques, E. T.; Xu, J.; August, J. T.
- Abstract
In our previous study by Gupta et al, dominant T-cell epitopes of SARS CoV-N(N) protein were predicted by software. The spectrum of interferon (IFN)-γ responses of Balb/c mice immunized against two different forms of SARS CoV-N plasmid was then analyzed. A cluster of dominant T-cell epitopes of SARS CoV-N protein was found in the N-terminus (amino acids 76–114). On the basis of this study, four different plasmids were constructed: (i) DNA encoding the unmodified N (p-N) or N70−122 (p-N70−122) as an endogenous cytoplasmic protein or (ii) DNA encoding a lysosome-associated membrane protein (LAMP) chimera with N (p-LAMP/N) or N70−122 (p-LAMP/N70−122). The immune responses of mice to these four constructs were evaluated. The results showed marked differences in the responses of the immunized mice. A single priming immunization with the p-LAMP/N construct was sufficient to elicit an antibody response. Enzyme-linked immunospot (ELISpot) assay indicated that p-LAMP/N70−122 and p-LAMP/N plasmids both elicited a greater IFN-γ response than p-N. p-N and p-N70−122 constructs induced low or undetectable levels of cytokine secretion. We also found that the p-LAMP/N70−122 construct promoted a long-lasting T-cell memory response without an additional boost 6 months after three immunizations. These findings show that DNA vaccines, even epitope-based DNA vaccines using LAMP as chimera, can elicit both humoral and cellular immune responses.
- Subjects
T cells; MEMBRANE proteins; CELLULAR immunity; DNA vaccines; CYTOPLASMIC inheritance
- Publication
Gene Therapy, 2009, Vol 16, Issue 11, p1353
- ISSN
0969-7128
- Publication type
Article
- DOI
10.1038/gt.2009.92