TABLE 1.
Description of explant media and p24 methods used for individual laboratories and VQA Lab
| Lab code (reference) | Tissue type | Polarized | Tissue size (mm)a | Base mediumb | Serum or additivesc | Antibiotic(s)d | p24 assay kit |
|---|---|---|---|---|---|---|---|
| VQA Labe | NA | NA | NA | R | 20% FBS, GL | Gent, Amp | Beckman Coulter |
| A (9) | Cervical | No | 3 × 3 × 2 | R | 10% FBS, GL | PS | Beckman Coulter |
| B (20) | Cervical | No | 3 × 3 × 2 | D | 10% HuAB, GL | PS | Perkin Elmer |
| C (4) | Stimulated cervical | Yes | 5 × 5 × 3 | D | 10% HuAB, 100 U/ml IL-2 | PS | NCIf |
| D (15) | Tonsil | No | 2 × 2 × 2 | R | 15% FBS, GL, N | Gent, F | Beckman Coulter (kinetic) |
| E (11) | Cervical | No | 3 × 3 × 3 | R | 20% FBS, GL | Gent, Amp | Zeptometrix |
| F (8) | Rectal | No | 3 × 3 × 2 | R | 10% FBS, GL | Z, F | NCIf |
| G (2, 16, 17)g | Cervical (PBMC coculture) | Yes | 5 × 5 × 3 | R | 20% FBS, GL | PS | Perkin Elmer |
Presented as length by width by height. NA, not applicable.
R, RPMI 1640; D, Dulbecco's modified Eagle medium.
GL, 2 to 4 mM l-glutamine; HuAB, human AB serum; N, 1× nonessential amino acids (100×); FBS, fetal bovine serum; IL-2, interleukin-2.
Gent, 50 μg/ml gentamicin sulfate; Amp, 50 μg/ml ampicillin; PS, 100 U/ml penicillin plus 100 μg/ml streptomycin; Z, 50 g/ml Zosyn; F, amphotericin B (Fungizone).
VQA Lab analyzed matched culture supernatants for p24 measurements from all laboratories and provided VQAM.
HIV-1 p24 antigen capture assay of AIDS Vaccine Program, NCI-Frederick Cancer Research and Development Center.
Laboratory G followed the published method, with the following modifications: HIV-1 transmission across the mucosa was determined by measuring HIV-1 p24 antigen in supernatants collected from PBMC cultures maintained in the bottom chamber of a transwell system.