Determination of neutralizing titers of polyclonal Ig purified from sera of infected individuals and of neutralizing monoclonal IgG using HIV-1BaL
. | Neutralizing titer, μg/mL . | . | . | . | |||
|---|---|---|---|---|---|---|---|
| . | . | Coculture . | . | . | |||
. | iMDDCs, purified at 99% . | iMDDCs, purified at 85% . | PBLs, purified at 15% . | PHA-stimulated PBMCs . | |||
| Polyclonal IgG | |||||||
| IgG no. 44 | 35 | 30 | 175 | 220 | |||
| IgG no. 8 | 40 | 30 | 250 | 155 | |||
| IgG no. 24 | 35 | 40 | 200 | 390 | |||
| IgG no. 23 | 60 | 45 | 400 | 600 | |||
| IgG no. 33 | 65 | ND | ND | 265 | |||
| IgG no. 36 | 35 | ND | ND | 350 | |||
| IgG no. 11 | 130 | 130 | — | — | |||
| IgG no. 3 | 350 | 195 | — | — | |||
| IgG control | — | — | — | — | |||
| Polyclonal IgA | |||||||
| IgA no. 8 | 215 | 290 | 250 | 360 | |||
| IgA no. 6 | 320 | 200 | 385 | 260 | |||
| IgA no. 35 | — | — | — | — | |||
| IgA control | — | — | — | — | |||
| Monoclonal IgG | |||||||
| IgG1 b12 | 1 | 3 | 15 | 25 | |||
| 2F5 IgG1k | 2 | 4 | 50 | 30 | |||
| 447-52D IgG3 | 3 | 3 | 50 | 50 | |||
| 2G12 IgG1k | 1 | 0.5 | ND | 50 | |||
| 4E10 IgG1 | 5 | ND | ND | 50 | |||
| Recombinant | |||||||
| sCD4 | 5 | 7 | 10 | 10 | |||
. | Neutralizing titer, μg/mL . | . | . | . | |||
|---|---|---|---|---|---|---|---|
| . | . | Coculture . | . | . | |||
. | iMDDCs, purified at 99% . | iMDDCs, purified at 85% . | PBLs, purified at 15% . | PHA-stimulated PBMCs . | |||
| Polyclonal IgG | |||||||
| IgG no. 44 | 35 | 30 | 175 | 220 | |||
| IgG no. 8 | 40 | 30 | 250 | 155 | |||
| IgG no. 24 | 35 | 40 | 200 | 390 | |||
| IgG no. 23 | 60 | 45 | 400 | 600 | |||
| IgG no. 33 | 65 | ND | ND | 265 | |||
| IgG no. 36 | 35 | ND | ND | 350 | |||
| IgG no. 11 | 130 | 130 | — | — | |||
| IgG no. 3 | 350 | 195 | — | — | |||
| IgG control | — | — | — | — | |||
| Polyclonal IgA | |||||||
| IgA no. 8 | 215 | 290 | 250 | 360 | |||
| IgA no. 6 | 320 | 200 | 385 | 260 | |||
| IgA no. 35 | — | — | — | — | |||
| IgA control | — | — | — | — | |||
| Monoclonal IgG | |||||||
| IgG1 b12 | 1 | 3 | 15 | 25 | |||
| 2F5 IgG1k | 2 | 4 | 50 | 30 | |||
| 447-52D IgG3 | 3 | 3 | 50 | 50 | |||
| 2G12 IgG1k | 1 | 0.5 | ND | 50 | |||
| 4E10 IgG1 | 5 | ND | ND | 50 | |||
| Recombinant | |||||||
| sCD4 | 5 | 7 | 10 | 10 | |||
Purified HIV-1BaL was preincubated with serial concentrations of polyclonal Ig purified from sera of HIV-1 patients or of neutralizing monoclonal IgG. Neutralizing titers that correspond to the concentration of Ig able to reduce the percentage of infected cells by 90% were determined for each cellular subset (purified iMDDCs, coculture of iMDDCs with 15% of PBLs, or PHA-stimulated PBMCs). Data are mean of 4 experiments performed with cells from at least 4 different donors.
— indicates no neutralizing activity detected at 800 μg/mL polyclonal Ig; ND, not determined.