Supplemental Figure Legends
Fig. S1 Relative levels of anti-gp140 Env antibody isotypes The levels of anti-gp140 specific IgG1, IgG2, IgG3, IgG4, IgA and IgM antibodies were compared in terms of µg/ml antibody equivalents relative to a standard concentration curve. Medians and percentiles with error bars are shown. The 10th, 25th, 75th and 90th percentile are shown as vertical lines. *p-value < 0.05, for comparison of IgA with IgG2 and IgG4. **p-value <0.001 for comparison of IgG1 with all antibody types
Fig. S2. Neutralizing activity of IgG eluted from gp120 WT and gp120 D368R mutant bead adsorption of plasma 1686. Neutralization of the JR-FL isolate by graded doses of IgG eluted from gp120 beads was determined by the TZM-bl assay.
Fig. S3. Alignment of MPER sequences. Sequences of the YU2 and 7312A MPER sequences and various 7312A chimeras with YU2 sequences engrafted as replacements are depicted (mutants C1-C8). Shading depicts residues that differ between the HIV-2 parent and the engrafted sequences. The sequences of 3 MPER peptides used in interference neutralization assays are also shown.
Fig. S4. Neutralization of HIV-2 MPER graft mutants. Plasma ID50 titers were determined against MPER graft mutants depicted in Fig. S3. The IC50s of MPER mAbs 2F5, 4E10 and Z13e1 were also measured, expressed in µg/ml. Neutralization against the JR-FL parent virus in the CF2 assay in the standard format is shown for reference purposes. Boxes are color coded in a similar manner as in Fig. 1. A summary of results from MPER peptide interference assays, derived from Fig. S5 is shown for comparison (column 10).
Fig. S5. Summary of MPER peptide interference of subtype B plasma neutralization. Neutralization interference of HXB2 and 7312A C1 chimeric virus by the MPER peptides depicted in Fig. S3 was determined. Data are recorded as fold decreases in ID50 titer. Incidences where the difference in titer is twofold or greater are shaded. The effects of peptides on MPER mAb IC50s 2F5, 4E10 and Z13e1 are included as controls. n.d. = not done.
Fig. S6. Alignment of V3 and C3 sequences. The V3 sequence of the HIV-2 KR parent and chimeras that bear the V3 sequences of YU2 or a consensus subtype C V3 are shown. Subtype B and C peptides from the N- (V3.01) or C- (V3.02) portions of the V3 loop are depicted. Similarly, subtype C C3 peptides from the N- (C3.01) or C- (C3.02) are delineated. Shading is used to emphasize variations from the parental V3 sequence and variable residues.
Fig. S7. Summary of CD4i activity. CD4i activity was measured by inhibition of virus capture by the E51 mAb (column 2) and by neutralization of HIV-2 7312A virus (column 3; background 7312A neutralizing activity in the standard format was subtracted), each in the presence of sCD4. Boxes are color coded with warmer colors reflecting higher titers.
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