Article Figures & Data
Figures
- FIG. 1.
Sequence conservation of the HIV-1 gp120 V3 loop (A) and the membrane-proximal gp41 ectodomain (C). Mutations were introduced in the V3 loop (B) and the membrane-proximal gp41 ectodomain (D) of SIV239 in order to generate the HIV-1 epitope-engrafted mutants.
- FIG. 2.
Comparative infectivity of SIV239, SIV316, SIV239-2F5, SIV239-4E10, SIV239-447D, SIV239-2F5-4E10, and SIV239-2F5-4E10-447D. Virus stocks were obtained from the transfection of 293T cells. Stocks were normalized to the amount of p27 and used to infect C8166-45 SIV-SEAP cells. (A) SEAP activity in supernatant was measured by using a chemiluminescent assay (see Materials and Methods) at 3 days postinfection. (B) SEAP activity normalized to the amount of p27. Neg., negative.
- FIG. 3.
Replication kinetics of viruses with the engrafted 2F5 and 4E10 HIV-1 epitopes in PBMCs obtained from two different rhesus macaques. (A) 419.91. (B) 382.90 d.p.i., days postinfection.
- FIG. 4.
Envelope incorporation into virions. Virions were pelleted, lysed, separated by SDS-PAGE, and visualized by Western blotting. (A) Western blot of viral pellets with anti-gp120 MAb and anti-p27 (capsid) MAb. (B) Relative Env incorporation into virions. The ratios of gp120 to p27 were calculated by using phosphorimaging analysis. Data are presented relative to the ratios found for SIV239 virions (Rn/R239).
- FIG. 5.
Comparative neutralization of SIV239, SIV239-2F5, and SIV239-4E10 by 4E10 HIV-1 monoclonal antibody (A) and 2F5 HIV-1 monoclonal antibody (B).
- FIG. 6.
Comparative neutralization of SIV239, SIV239-2F5, and SIV239-4E10 by pooled plasma from six SIVmac-positive rhesus macaques (top row), different rhesus anti-gp120 MAbs (middle row), and a mouse anti-gp41 MAb (bottom row). Neg., negative.
- FIG. 7.
Peptide competition. The possible correlation between 4E10-specific antibodies and neutralization by LTNP2 plasma was assessed by competition with peptide 94-1 (A). The ability of the 94-1 peptide to bind to the antigen-binding site of 4E10 antibody was also corroborated in a competition neutralization assay (B).
- FIG. 8.
Neutralization of 239-4E10 by LTNP2 plasma (A) and IgG and IgA isolated from LTNP2 plasma (B).
Tables
- TABLE 1.
Neutralization of four viruses by 92 HIV-positive plasma samplesa
↵ a Pooled rhesus plasma from SIV-negative animals served as a negative control. The viruses are organized horizontally, and the plasma samples are organized from top to bottom. Dates refer to time that the sample was collected. 1/20 and 1/200 indicate 20- and 200-fold dilutions of plasma, respectively. Numbers indicate percent SEAP activity (counts per second ± the standard deviation). A white box indicates >50% SEAP activity, a yellow box indicates <50% SEAP activity but >10% SEAP activity, and a red box indicates <10% SEAP activity.
- TABLE 2.
Percentage of SEAP activity in the peptide competition neutralization assaya
Patient Date (mo/day/yr) % of SEAP activity Neut. 1/20b Neut. 1/20+94Ic d381 3/1/05 56 ± 14 50 ± 6 i447 2/17/05 57 ± 4 52 ± 11 VI2992 8/20/04 49 ± 6 64 ± 13 ↵ a Plasmas with moderate neutralization activity against SIV239-4E10 (>50% <90%) were competed with peptide 94-1 as indicated in Materials and Methods.
↵ b Percentage of SEAP activity ± standard deviation after neutralization of SIV239-4E10 with the corresponding plasma.
↵ c Percentage of SEAP activity ± standard deviation after preincubation of each plasma with peptide 94-1 followed by neutralization of virus SIV239-4E10.
