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- This epitope is located in the V2 region of UG92005 (UG, clade D) and the hybridoma that recognized it used Vβ5.
- 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 IAb 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 IAb 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.
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.
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