Construction and immunogenicity study of a 297-bp humanized HIV V3 DNA of an approximated last common ancestor in mice.
Asian Pac J Allergy Immunol
;
2004 Mar; 22(1): 49-60
Article
in English
| IMSEAR
| ID: sea-36481
ABSTRACT
DNA immunization represents one of the promising HIV-1 vaccine approaches. To overcome the obstacle of genetic variation, we used the last common ancestor (LCA) or "center-of-the-tree" approach to study a DNA fragment of the HIV-1 envelope surrounding the V3 region. A humanized codon of the 297-bp consensus ancestral sequence of the HIV-1 envelope (codons 291-391) was derived from the 80 most recent HIV-1 isolates from the 8 circulating HIV-1 subtypes worldwide. This 297-bp humanized "multi-clade" V3 DNA was amplified by a PCR-based technique. The PCR product was well expressed in vitro whereas the corresponding non-humanized V3 DNA (subtype A/E) could not be expressed. However, both V3 DNA constructs as well as the full-length HIV-1 envelope construct (A/E) were found to be immunogenic in mice by the footpad-swelling assay. Moreover, intracellular and extracellular interferon-gamma could be detected upon in vitro stimulation of spleen cells although the response was relatively weak. Further improvement of our humanized V3 DNA is needed.
Full text:
Available
Index:
IMSEAR (South-East Asia)
Main subject:
Female
/
DNA, Viral
/
Viral Envelope Proteins
/
Polymerase Chain Reaction
/
HIV-1
/
AIDS Vaccines
/
Vaccines, DNA
/
Models, Animal
/
Animals
/
Mice
Language:
English
Journal:
Asian Pac J Allergy Immunol
Year:
2004
Type:
Article
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