TABLE 2.
Relative fitnesses of wild-type and site-directed mutant HIV-1xxLAI in growth competition assays
| Competition condition and genotype of competing viruses | Relative fitness valuea | P valueb | Fitness interpretation |
|---|---|---|---|
| Without drug | |||
| WT vs WTc | 1.02 ± 0.02 | WT = WT | |
| WT vs RT-M184V | 0.84 ± 0.08 | 0.008 | WT > RT-M184V |
| WT vs IN-N155H | 0.71 ± 0.08 | 0.001 | WT > IN-N155H |
| WT vs IN-Q148R | 0.71 ± 0.03 | <0.001 | WT > IN-Q148R |
| WT vs RT-M184V + IN-N155H | 0.66 ± 0.1 | <0.001 | WT > RT-M184V + IN-N155H |
| WT vs RT-M184V + IN-Q148R | 0.60 ± 0.1 | <0.001 | WT > RT-M184V + IN-Q148R |
| WT vs RT-K65R/M184V + IN-Q148R | < 0.50 ± 0.1 | <0.001 | WT > RT-K65R/M184V + IN-Q148R |
| WT vs RT-K65R/M184V + IN-N155H | < 0.50 ± 0.2 | <0.001 | WT > RT-K65R/M184V + IN-N155H |
| IN-N155H vs IN-Q148R | 0.97 ± 0.06 | 0.164 | IN-N155H ≈ IN-Q148R |
| RT-M184V vs RT-M184V + IN-Q148R | 0.75 ± 0.05 | <0.001 | RT-M184V > RT-M184V + IN-Q148R |
| RT-M184V vs RT-M184V + IN-N155H | 0.76 ± 0.01 | <0.001 | RT-M184V > RT-M184V + IN-N155H |
| IN-Q148R vs RT-M184V + IN-Q148R | 0.93 ± 0.09 | 0.111 | IN-Q148R ≥ RT-M184V + IN-Q148R |
| IN-N155H vs RT-M184V + IN-N155H | 0.94 ± 0.07 | 0.091 | IN-N155H ≥ RT-M184V + IN-N155H |
| With drug | |||
| WT vs IN-Q148R (0.5 nM EVG) | 0.76 ± 0.02 | <0.001 | WT > IN-Q148R |
| WT vs IN-Q148R (2 nM EVG) | 0.95 ± 0.04 | 0.038 | WT ≈ IN-Q148R |
| WT vs IN-Q148R (10 nM EVG) | 2.09 ± 0.1 | <0.001 | WT < IN-Q148R |
| WT vs IN-N155H (0.5 nM EVG) | 0.85 ± 0.04 | 0.001 | WT > IN-N155H |
| WT vs IN-N155H (2 nM EVG) | 1.10 ± 0.05 | 0.043 | WT ≤ IN-N155H |
| WT vs IN-N155H (10 nM EVG) | 2.03 ± 0.5 | 0.010 | WT < IN-N155H |
| IN-N155H vs IN-Q148R (1 nM EVG) | 1.07 ± 0.06 | 0.178 | IN-N155H ≤ IN-Q148R |
| IN-N155H vs IN-Q148R (10 nM EVG) | 0.83 ± 0.05 | 0.001 | IN-N155H > IN-Q148R |
| IN-N155H vs IN-Q148R (50 nM EVG) | 1.32 ± 0.4 | 0.194 | IN-N155H ≤ IN-Q148R |
| IN-N155H vs IN-Q148R (100 nM EVG) | 1.77 ± 0.1 | <0.001 | IN-N155H < IN-Q148R |
| RT-M184V vs RT-M184V + IN-Q148R (0.5 nM EVG, 1 nM FTC) | 0.82 ± 0.05 | 0.001 | RT-M184V > RT-M184V + IN-Q148R |
| RT-M184V vs RT-M184V + IN-Q148R (10 nM EVG, 100 nM FTC) | 1.38 ± 0.09 | 0.001 | RT-M184V < RT-M184V + IN-Q148R |
| RT-M184V vs RT-M184V + IN-N155H (0.5 nM EVG, 1 nM FTC) | 0.84 ± 0.1 | 0.024 | RT-M184V ≥ RT-M184V + IN-N155H |
| RT-M184V vs RT-M184V + IN-N155H (10 nM EVG, 100 nM FTC) | 1.57 ± 0.2 | 0.006 | RT-M184V < RT-M184V + IN-N155H |
| IN-Q148R vs RT-M184V + IN-Q148R (1 nM EVG, 1 nM FTC) | 0.83 ± 0.1 | 0.037 | IN-Q148R ≥ RT-M184V + IN-Q148R |
| IN-Q148R vs RT-M184V + IN-Q148R (100 nM EVG, 100 nM FTC) | 1.61 ± 0.3 | 0.010 | IN-Q148R < RT-M184V + IN-Q148R |
| IN-N155H vs RT-M184V + IN-N155H (1 nM EVG, 1 nM FTC) | 0.86 ± 0.1 | 0.029 | IN-N155H ≥ RT-M184V + IN-N155H |
| IN-N155H vs RT-M184V + IN-N155H (100 nM EVG, 100 nM FTC) | 1.38 ± 0.1 | 0.003 | IN-N155H < RT-M184V + IN-N155H |
The relative fitness (RF) value of the mutant in competition with the wild type was calculated as follows: (1 + s) = exp {(1/t) × ln[(Mt/Wt) × (Mt0/Wt0)]}, where s is the selection coefficient; t is the time (in days); Mt and Mt0 are the fractions of mutant virus initially and at the time of measurement, respectively; and Wt and Wt0 are the fractions of wild-type virus initially and at the time of measurement, respectively (27). Mutant-versus-mutant competitions were analyzed using the same equation. The data represent the means and standard deviations from at least 3 independent experiments.
P values were determined using a two-tailed Student's t test comparing the competitions to the wild-type-versus-wild-type competition.
A control experiment was performed to verify that isogenic HIV-1 recombinants differing only in their sequence tags would grow with equivalent fitnesses. WT, wild type.