Immune responses to T-cell epitopes of SARS CoV-N protein are enhanced by N immunization with a chimera of lysosome-associated membrane protein

K. Yang, K. Sun, K. N. Srinivasan, J. Salmon, E. T. Marques, J. Xu, Thomas August

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)1353-1362
Number of pages10
JournalGene Therapy
Volume16
Issue number11
DOIs
StatePublished - 2009

Fingerprint

Lysosome-Associated Membrane Glycoproteins
T-Lymphocyte Epitopes
Immunization
SARS Virus
DNA Vaccines
Plasmids
Interferons
Enzyme-Linked Immunospot Assay
Coronavirus nucleocapsid protein
DNA
Humoral Immunity
Cellular Immunity
Antibody Formation
Epitopes
Proteins
Software
Cytokines
T-Lymphocytes
Amino Acids

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

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Immune responses to T-cell epitopes of SARS CoV-N protein are enhanced by N immunization with a chimera of lysosome-associated membrane protein. / Yang, K.; Sun, K.; Srinivasan, K. N.; Salmon, J.; Marques, E. T.; Xu, J.; August, Thomas.

In: Gene Therapy, Vol. 16, No. 11, 2009, p. 1353-1362.

Research output: Contribution to journalArticle

Yang, K. ; Sun, K. ; Srinivasan, K. N. ; Salmon, J. ; Marques, E. T. ; Xu, J. ; August, Thomas. / Immune responses to T-cell epitopes of SARS CoV-N protein are enhanced by N immunization with a chimera of lysosome-associated membrane protein. In: Gene Therapy. 2009 ; Vol. 16, No. 11. pp. 1353-1362.
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