Abstract
The immune responses to an HIV-1 p55Gag vaccine encoded as a DNA chimera with the lysosomal associated membrane protein-1 (LAMP) have been examined for the effect of the addition of the inverted terminal repeat (ITR) sequences of the adeno-associated virus (AAV) to the DNA plasmid construct, and of packaging the LAMP/gag gene as a recombinant AAV vector (rAAV). DNA plasmids encoding Gag and the LAMP/Gag protein chimera were constructed in two vectors, the pcDNA3.1 and a corresponding plasmid containing the ITR sequences (pITR) flanking the expression elements of the plasmid, and the pITR LAMP/gag DNA plasmid was encapsidated in the rAAV vector. Human 293 cells transfected in vitro with LAMP/gag plasmids either in pcDNA3.1 or pITR produced much Gag protein in cell extracts (1.6 and 2.2 ng of Gag/mg of protein, respectively). The immune responses of mice to immunization with these constructs were examined under three protocols: DNA prime/DNA boost, DNA prime/rAAV boost, and a single rAAV immunization. The results demonstrated that under DNA prime/DNA boost protocol, the "naked" DNA vaccines encoding the LAMP/gag chimera, either as pcDNA3.1 or pITR DNA plasmid constructs, elicited strong CD4+ T cell responses. In contrast, significantly higher levels of CD8+ and antibody responses were observed with the pITR-DNA constructs. Immunization with the rAAV vector under the DNA prime/rAAV boost protocol resulted in sustained T cell responses and a markedly increased antibody response, predominantly of the IgG1 isotype resulting from the activation of the Th2 subset of CD4+ T cells, that was sustained for at least 5 months after immunization.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 220-232 |
| Number of pages | 13 |
| Journal | Virology |
| Volume | 323 |
| Issue number | 2 |
| DOIs | |
| State | Published - Jun 1 2004 |
Fingerprint
Keywords
- AAV
- antigen presenting cell
- APC
- CD4
- CD8
- DNA vaccine
- HIV-1 Gag
- Inverted terminal repeat
- LAMP
- lysosome-associated membrane protein
- major histocompatibility complex
- MHC
- MHC class II-enriched compartment
- MHC II
- MIIC
- rAAV
- Rev-responsive elements
- RRE
ASJC Scopus subject areas
- Virology
- Infectious Diseases
Cite this
Inverted terminal repeat sequences of adeno-associated virus enhance the antibody and CD8+ responses to a HIV-1 p55Gag/LAMP DNA vaccine chimera. / Chikhlikar, Priya; Barros De Arruda, Luciana; Agrawal, Shikha; Byrne, Barry; Guggino, William B; August, Thomas; Marques, Ernesto T A.
In: Virology, Vol. 323, No. 2, 01.06.2004, p. 220-232.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Inverted terminal repeat sequences of adeno-associated virus enhance the antibody and CD8+ responses to a HIV-1 p55Gag/LAMP DNA vaccine chimera
AU - Chikhlikar, Priya
AU - Barros De Arruda, Luciana
AU - Agrawal, Shikha
AU - Byrne, Barry
AU - Guggino, William B
AU - August, Thomas
AU - Marques, Ernesto T A
PY - 2004/6/1
Y1 - 2004/6/1
N2 - The immune responses to an HIV-1 p55Gag vaccine encoded as a DNA chimera with the lysosomal associated membrane protein-1 (LAMP) have been examined for the effect of the addition of the inverted terminal repeat (ITR) sequences of the adeno-associated virus (AAV) to the DNA plasmid construct, and of packaging the LAMP/gag gene as a recombinant AAV vector (rAAV). DNA plasmids encoding Gag and the LAMP/Gag protein chimera were constructed in two vectors, the pcDNA3.1 and a corresponding plasmid containing the ITR sequences (pITR) flanking the expression elements of the plasmid, and the pITR LAMP/gag DNA plasmid was encapsidated in the rAAV vector. Human 293 cells transfected in vitro with LAMP/gag plasmids either in pcDNA3.1 or pITR produced much Gag protein in cell extracts (1.6 and 2.2 ng of Gag/mg of protein, respectively). The immune responses of mice to immunization with these constructs were examined under three protocols: DNA prime/DNA boost, DNA prime/rAAV boost, and a single rAAV immunization. The results demonstrated that under DNA prime/DNA boost protocol, the "naked" DNA vaccines encoding the LAMP/gag chimera, either as pcDNA3.1 or pITR DNA plasmid constructs, elicited strong CD4+ T cell responses. In contrast, significantly higher levels of CD8+ and antibody responses were observed with the pITR-DNA constructs. Immunization with the rAAV vector under the DNA prime/rAAV boost protocol resulted in sustained T cell responses and a markedly increased antibody response, predominantly of the IgG1 isotype resulting from the activation of the Th2 subset of CD4+ T cells, that was sustained for at least 5 months after immunization.
AB - The immune responses to an HIV-1 p55Gag vaccine encoded as a DNA chimera with the lysosomal associated membrane protein-1 (LAMP) have been examined for the effect of the addition of the inverted terminal repeat (ITR) sequences of the adeno-associated virus (AAV) to the DNA plasmid construct, and of packaging the LAMP/gag gene as a recombinant AAV vector (rAAV). DNA plasmids encoding Gag and the LAMP/Gag protein chimera were constructed in two vectors, the pcDNA3.1 and a corresponding plasmid containing the ITR sequences (pITR) flanking the expression elements of the plasmid, and the pITR LAMP/gag DNA plasmid was encapsidated in the rAAV vector. Human 293 cells transfected in vitro with LAMP/gag plasmids either in pcDNA3.1 or pITR produced much Gag protein in cell extracts (1.6 and 2.2 ng of Gag/mg of protein, respectively). The immune responses of mice to immunization with these constructs were examined under three protocols: DNA prime/DNA boost, DNA prime/rAAV boost, and a single rAAV immunization. The results demonstrated that under DNA prime/DNA boost protocol, the "naked" DNA vaccines encoding the LAMP/gag chimera, either as pcDNA3.1 or pITR DNA plasmid constructs, elicited strong CD4+ T cell responses. In contrast, significantly higher levels of CD8+ and antibody responses were observed with the pITR-DNA constructs. Immunization with the rAAV vector under the DNA prime/rAAV boost protocol resulted in sustained T cell responses and a markedly increased antibody response, predominantly of the IgG1 isotype resulting from the activation of the Th2 subset of CD4+ T cells, that was sustained for at least 5 months after immunization.
KW - AAV
KW - antigen presenting cell
KW - APC
KW - CD4
KW - CD8
KW - DNA vaccine
KW - HIV-1 Gag
KW - Inverted terminal repeat
KW - LAMP
KW - lysosome-associated membrane protein
KW - major histocompatibility complex
KW - MHC
KW - MHC class II-enriched compartment
KW - MHC II
KW - MIIC
KW - rAAV
KW - Rev-responsive elements
KW - RRE
UR - http://www.scopus.com/inward/record.url?scp=2942532790&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=2942532790&partnerID=8YFLogxK
U2 - 10.1016/j.virol.2004.02.025
DO - 10.1016/j.virol.2004.02.025
M3 - Article
C2 - 15193918
AN - SCOPUS:2942532790
VL - 323
SP - 220
EP - 232
JO - Virology
JF - Virology
SN - 0042-6822
IS - 2
ER -