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Construction and immunogenicity study of a 297-bp humanized HIV V3 DNA of an approximated last common ancestor in mice.

Sirivichayakul S, Tirawatnapong T, Ruxrungtham K, Oelrichs R, Lorenzen SL, Xin KQ, Okuda K, Phanuphak P

  • Journal Asian Pacific journal of allergy and immunology

  • Published 22 Feb 2005

  • Volume 22

  • ISSUE 1

  • Pagination 49-60


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.