Production of a highly cytopathic HIV-1 isolate from a human mucosal epithelial cell line cultured on microcarrier beads in serum-free medium.
The human colonic epithelial cell line HT-29 can be productively infected with various HIV-1 and HIV-2 isolates that are highly cytopathic for T lymphocytes. In each case, a chronically infected HT-29 cell line can be established, and progeny viruses retain their original properties including high cytopathogenicity for T cells. Inasmuch as AIDS vaccines should include viral isolates capable of infecting mucosal epithelial cells, it may be useful to produce these isolates in such cells at a large scale. We describe here a microcarrier-based culture system allowing the production of infectious viruses from HT-29 cells grown in a chemically defined serum-free medium (Dulbecco's modified Eagle's medium/F12, HEPES 15 mM, pH 7.4, transferrin 5 micrograms/ml, selenium 10 ng/ml). The yield of HIV-1 from microcarrier cultures (275 ng of p24gag/ml) was greater than the yield from conventional culture flasks (122 ng of p24gag/ml). This virus, produced in serum-free medium, can be used either as a viral stock or as a source for HIV-1 proteins.
Yahi
N
N
CNRS URA 1455, Faculté de Médecine Nord, Marseille, France.
Baghdiguian
S
S
Fantini
J
J
D000230:Adenocarcinoma; D003110:Colonic Neoplasms; D003470:Culture Media; D004848:Epithelium / Q000821:virology; D005455:Fluorescent Antibody Technique; D016655:HIV Core Protein p24 / Q000032:analysis; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000254:growth & development*; D006801:Humans; D007413:Intestinal Mucosa / Q000821:virology*; D008854:Microscopy, Electron; D008863:Microspheres; D012194:RNA-Directed DNA Polymerase / Q000032:analysis; D012643:Selenium / Q000494:pharmacology; D014168:Transferrin / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014776:Virus Cultivation / Q000379:methods*
D003470:Culture Media; D016655:HIV Core Protein p24; D014168:Transferrin; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase; D012643:Selenium