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HXB2 Location	gp160(611-620) gp41(100-109) DNA(8055..8084)	gp160 Epitope Map
Author Location	gp41( 1007, UG92005)
Epitope	NASWSNKSLE	Epitope Alignment
Species (MHC/HLA)	mouse(H-2 IAb)
Immunogen	vaccine
Experimental methods
Keywords	subtype comparisons, epitope processing, TCR usage

Vaccine Details

Notes

• This gp41 epitope is conserved in 1007 (US, clade B) and UG92005 (UG, clade D) and was recognized by two hybridomas from two different mice that were vaccinated with different clades -- the Vβ usage was Vβ 4 and 14.
• The epitope described here is the region of overlap of two 15 mers that were both able to stimulate IL-2 production from the hybridoma (T[TN]VPWNASWSNKSLE and NASWSNKSLEQIWNN) -- the only difference between 1007 and UG92005 for these two proteins is that 1007 has a T and UG92005 has an N in the second position of the first peptide.
• C57BL/6 mice were immunized with a prime-boost strategy involving three HIV-1 Env antigens: Mice were primed with DNA given i.m., 3-4 weeks later boosted with VV, and 3-4 weeks later boosted again with purified protein in Freund's adjuvant.
• The vaccinia construct is a pSC11-based VV vector with the first 38 amino acids contributed by BH10 and the rest of gp120 and gp41 by the vaccine strain, the DNA construct is in the pJW4303 vector with a CMV promotor, and the purified protein is expressed from the pJW4303 vector transfected into CHO-K1 cells.
• Ten days after the final boost, hybridomas were made and tested for IL-2 production using either B6 spleen cells or H-2 IA^b transfected L cells as targets and Vβ usage was determined.
• Mice were immunized with an Env from either one of two clades: HIV-1 1007, a clade B strain isolated from an individual from Memphis Tennesee, and HIV-1 92UG005, a clade D vaccine isolated from Uganda in 1992 through the WHO.
• 80 unique clonotypes were characterized from six mice.
• H-2 IA^b restricted T-helper epitopes were concentrated in 5 distinct regions within the Env sequence (2 clonotype responses in V2, 26 in C2, 22 in V3, 23 in V4C4, and 7 in gp41).
• Epitope hotspots tended to be proximal to heavily glycosylated regions of the Env sequence, in exposed, non-helical strands of the protein. The non-uniform localization may be influenced by differential antigen processing and the glycosylation site proximity may allow binding to lectins and promote trafficking through processing pathways.

References

Surman2001 S. Surman, T. D. Lockey, K. S. Slobod, B. Jones, J. M. Riberdy, S. W. White, P. C. Doherty, and J. L. Hurwitz. Localization of CD4+ T Cell Epitope Hotspots to Exposed Strands of HIV Envelope Glycoprotein Suggests Structural Influences on Antigen Processing. Proc. Natl. Acad. Sci. U.S.A., 98(8):4587-4592, 10 Apr 2001. URL: http://www.pnas.org/cgi/content/full/98/8/4587. PubMed ID: 11287644. Show all entries for this paper.