TABLE 1.
MPN values for PG9 and functional stability (T90) of mutants of ADA and Comb-muta
| Strain | ADA |
Comb-mut |
Comment(s) | |||||||
|---|---|---|---|---|---|---|---|---|---|---|
| PG9 MPN (%) | t test (PG9 MPN) | T90 (°C) | t test (T90) | Infectivity (% WT) | 447-52D IC50 (μg/ml) | PG9 MPN (%) | t test | Infectivity (% WT) | ||
| Wild type | 63.6 ± 4.2 | 42.7 ± 0.6 | 100 ± 4.1 | 0.050 | 87.2 ± 3.5 | 100 ± 5.8 | ||||
| K46A | 88.6 ± 2.4 | **** | 44.5 ± 1.3 | NS | 103 ± 3.0 | ND | 86.0 ± 3.0 | NS | 86 ± 2.6 | Neutralization resistance in SIV (13) |
| E87A | 81.3 ± 5.8 | **** | 44.5 ± 1.3 | NS | 101 ± 8.5 | ND | 85.7 ± 3.3 | NS | 109 ± 1.4 | 5P12-RANTES escape (D. Mosier, unpublished) |
| L111A | 80.2 ± 1.8 | **** | 44.7 ± 0.5 | NS | 119 ± 4.7 | 0.13 | 79.1 ± 0.6 | NS | 16 ± 0.6 | Reduced gp120 dimers (88, 89) |
| K121Q | ND | ND | ND | ND | 86.8 ± 3.8 | NS | 36 ± 7.2 | Structural analysisb | ||
| N130Q | 29.2 ± 8.1 | **** | 42.8 ± 0.8 | NS | 101 ± 0.5 | ND | ND | ND | Sequence alignmentc | |
| D133N | 61.0 ± 6.0 | NS | 42.6 ± 0.2 | NS | 132 ± 1.6 | 0.043 | ND | ND | Sequence alignment | |
| R135T | 69.3 ± 3.3 | NS | 44.4 ± 1.4 | NS | 93 ± 15.0 | ND | 76.5 ± 3.3 | ** | 28 ± 0.9 | Sequence alignment |
| N136S | 77.6 ± 3.6 | **** | 47.3 ± 0.4 | **** | 106 ± 2.5 | 4.7 | 91.6 ± 2.6 | NS | 89 ± 1.1 | 5P12-RANTES escape (D. Mosier, unpublished) |
| V137N | 61.0 ± 8.4 | NS | 42.4 ± 0.2 | NS | 36 ± 1.6 | 0.022 | ND | ND | Sequence alignment | |
| S143D | 79.0 ± 3.6 | **** | 46.5 ± 1.2 | *** | 96 ± 2.9 | 6.4 | 81.8 ± 6.8 | NS | 43 ± 2.6 | Sequence alignment |
| I153L | 65.6 ± 8.6 | NS | 42.3 ± 0.6 | NS | 66 ± 0.4 | ND | ND | ND | Sequence alignment | |
| I161M | 59.5 ± 3.8 | NS | 41.8 ± 1.1 | NS | 48 ± 2.0 | 0.00064 | ND | ND | Sequence alignment | |
| S164E | 77.3 ± 0.5 | *** | 45.0 ± 0.8 | NS | 33 ± 1.6 | 0.025 | 89.4 ± 4.8 | NS | 71 ± 3.5 | Sequence alignment |
| I165L | 80.1 ± 2.7 | **** | 43.4 ± 1.4 | NS | 78 ± 2.0 | 0.0085 | 94.7 ± 1.6 | NS | 81 ± 6.1 | Sequence alignment |
| V169K | 94.7 ± 3.2 | **** | 43.0 ± 1.0 | NS | 90 ± 7.7 | 0.31 | 92.6 ± 4.2 | NS | 102 ± 3.9 | Correlate with RV144 protection (63) |
| K170Q | 26.0 ± 8.5 | **** | 42.5 ± 0.5 | NS | 110 ± 3.8 | 10.3 | 47.2 ± 2.7 | **** | 86 ± 0.7 | Sequence alignment |
| D172V | 59.4 ± 4.2 | NS | 43.7 ± 1.1 | NS | 59 ± 1.7 | ND | 69.9 ± 3.2 | *** | 103 ± 6.1 | Sequence alignment |
| A174S | 30.2 ± 7.1 | **** | 39.8 ± 0.4 | NS | 20 ± 0.8 | 0.00028 | ND | ND | Sequence alignment | |
| P183Q | ND | ND | ND | ND | 86.7 ± 4.4 | NS | 67 ± 6.2 | Structural analysis | ||
| N186G D187S N188S | ND | ND | ND | ND | 88.8 ± 2.1 | NS | 70 ± 8.5 | Structural analysis | ||
| D187N | 80.9 ± 2.6 | **** | 44.0 ± 0.3 | NS | 77 ± 2.9 | 0.0088 | 89.1 ± 4.4 | NS | 56 ± 4.9 | Sequence alignment |
| T200A | 52.0 ± 4.9 | ** | 42.2 ± 0.5 | NS | 6 ± 0.5 | ND | ND | ND | Sequence alignment | |
| T219A | 58.8 ± 6.5 | NS | 42.6 ± 0.6 | NS | 90 ± 7.5 | 0.33 | ND | ND | Sequence alignment | |
| N276D | 72.0 ± 8.5 | * | 44.3 ± 0.4 | NS | 79 ± 4.4 | 31.7 | 87.2 ± 3.1 | NS | 112 ± 1.2 | Knocks out N-glycosylation site |
| N302Y | 78.0 ± 5.5 | **** | 45.3 ± 0.7 | * | 66 ± 3.2 | 7.3 | 86.4 ± 2.1 | NS | 74 ± 3.0 | 5P12-RANTES escape (D. Mosier, unpublished) |
| I309M | 37.0 ± 5.3 | **** | 42.6 ± 0.3 | NS | 6 ± 0.6 | 0.0010 | ND | ND | 5P12-RANTES escape (D. Mosier, unpublished) | |
| R315Q | 77.3 ± 4.1 | **** | 47.5 ± 0.2 | ** | 82 ± 5.4 | >100 | 83.0 ± 3.9 | NS | 114 ± 4.4 | Clade C V3 consensus (90) |
| A316T | 73.6 ± 1.9 | ** | 46.2 ± 1.7 | *** | 91 ± 4.5 | 55.5 | 83.2 ± 3.1 | NS | 108 ± 3.9 | Clade C V3 consensus (90) |
| A316W | ND | ND | ND | ND | 85.8 ± 2.9 | NS | 39 ± 2.4 | Structural analysis | ||
| F317W | 66.2 ± 6.0 | NS | 41.2 ± 0.5 | * | 12 ± 1.0 | 0.0034 | ND | ND | 5P12-RANTES escape (D. Mosier, unpublished) | |
| Y318F | 63.5 ± 3.0 | NS | 42.8 ± 0.9 | NS | 114 ± 6.1 | 0.036 | ND | ND | 5P12-RANTES escape (D. Mosier, unpublished) | |
| T319A | 66.2 ± 4.5 | NS | 46.2 ± 0.4 | ** | 116 ± 4.7 | 43.0 | 83.7 ± 1.3 | NS | 23 ± 1.7 | Clade C V3 consensus (90) |
| T320W | ND | ND | ND | ND | 87.7 ± 3.2 | NS | 130 ± 3.8 | Structural analysis | ||
| I420M | ND | ND | ND | ND | 86.4 ± 3.1 | NS | 68 ± 3.9 | Structural analysis | ||
| R440A | 85.9 ± 0.6 | **** | 44.8 ± 1.4 | NS | 102 ± 2.1 | 21.3 | 80.3 ± 3.8 | NS | 105 ± 5.1 | Structural analysis |
| R440Q | ND | ND | ND | ND | 86.8 ± 5.2 | NS | 87 ± 2.5 | Structural analysis | ||
| Q442V | ND | ND | ND | ND | 88.0 ± 4.5 | NS | 86 ± 2.3 | Structural analysis | ||
| N461G | 69.3 ± 3.8 | NS | 44.0 ± 0.5 | NS | 126 ± 16.1 | 1.3 | 82.4 ± 2.3 | NS | 97 ± 6.9 | Knocks out N-glycosylation site |
| N553S | 79.5 ± 4.2 | **** | 44.4 ± 1.5 | NS | 95 ± 5.7 | 0.59 | 83.4 ± 2.4 | NS | 97 ± 3.4 | Stabilized gp140 trimer (91) |
| I559Y | 64.6 ± 2.9 | NS | 44.9 ± 0.6 | NS | 19 ± 2.3 | ND | 67.7 ± 2.9 | **** | 11 ± 0.6 | Structural analysis |
| L565M | 61.0 ± 3.3 | NS | 46.0 ± 0.4 | ** | 104 ± 8.7 | 1.2 | ND | 0 ± 0.0 | 5P12-RANTES escape (D. Mosier, unpublished) | |
| Q567K | 71.4 ± 2.4 | * | 46.3 ± 0.3 | ** | 76 ± 4.7 | 64.7 | 84.0 ± 2.0 | NS | 82 ± 4.2 | Stabilized gp140 trimer (91) |
| T605P | 78.4 ± 3.9 | **** | 47.7 ± 1.2 | **** | 103 ± 3.7 | 12.6 | 85.8 ± 2.3 | NS | 70 ± 5.0 | Identified from drug screen (DPL) |
| A612S | 64.6 ± 4.2 | NS | 44.7 ± 0.2 | NS | 95 ± 2.9 | 1.76 | 84.5 ± 3.8 | NS | 88 ± 1.6 | Identified from drug screen (DPL) |
a
PG9 maximum percent neutralization (MPN) was determined at 5 μg/ml. ND, not determined; NS, not significant. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. The “Infectivity (% WT)” columns indicate the percentages of wild-type virus infectivity ± the standard deviation.
b
These mutations in subunit interfaces were anticipated to impact Env trimer stability and conformation based on analysis of an X-ray crystal structure of BG505 SOSIP gp140 (8).
c
After sequence alignments, the amino acid residues in ADA/Comb-mut were changed to that of HIV-1 isolates BG505 or 16055, which are fully neutralized by PG9 and PG16.