Syncytium induction by fresh HIV isolates: quantitative analysis using a transactivation beta-gal assay.
We used a quantitative bioassay (the beta-gal assay) to visualize and quantify syncytium induction by fresh HIV isolates. This bioassay is based on the transactivation by tat of a chimeric gene comprising an HIV-1 long terminal repeat (LTR) fused to a modified lacZ gene of Escherichia coli. The chimeric gene encodes a beta-galactosidase which is translocated to the nucleus. It allows the enzymatic staining of all nuclei from HIV-induced syncytia. Using this unequivocal assay (the beta-gal assay), we could assess the syncytium-inducing properties of fresh HIV isolates after only 4 days of coculture of patient lymphocytes with activated normal lymphocytes. Syncytium-inducing HIV isolates were detected in 11 out of 40 seropositive patients studied. They were isolated mainly from AIDS patients: eight out of 17 grade IV (according to Centers for Disease Control criteria) patients were infected with syncytium-inducing strains. However, of 23 grade II and III patients tested, syncytium-inducing HIV strains were isolated from three cases. These three patients displayed no detectable p24 antigenaemia and had a CD4+ cell count of greater than 300 cells/microliter. The in vitro replication rate of HIV grown from 36 patient blood samples was then examined by sequential p24 antigen measurements in coculture supernatants. The 10 samples leading to syncytium formation also exhibited the highest replication rate. The possibility of unequivocally detecting syncytium-inducing strains after only a few days of coculture will make this detection routine and rapid. In addition, the limited period of amplification required is a significant advantage as it minimizes the emergence of HIV variants selected during long-term in vitro cultures.
Emilie
D
D
INSERM U131, Clamart, France.
Maillot
M C
MC
Bonnerot
C
C
Devergne
O
O
Delfraissy
J F
JF
Nicolas
J F
JF
Galanaud
P
P
D000163:Acquired Immunodeficiency Syndrome / Q000382:microbiology*; D015496:CD4-Positive T-Lymphocytes / Q000382:microbiology; D003588:Cytopathogenic Effect, Viral / Q000235:genetics / Q000276:immunology; D015683:Gene Products, gag / Q000276:immunology; D015488:HIV Antigens / Q000032:analysis; D016655:HIV Core Protein p24; D015497:HIV-1 / Q000235:genetics / Q000254:growth & development* / Q000276:immunology; D006801:Humans; D007958:Leukocyte Count; D012091:Repetitive Sequences, Nucleic Acid; D015533:Transcriptional Activation; D014758:Viral Core Proteins / Q000276:immunology; D014779:Virus Replication / Q000235:genetics / Q000276:immunology; D001616:beta-Galactosidase / Q000032:analysis*
D015683:Gene Products, gag; D015488:HIV Antigens; D016655:HIV Core Protein p24; D014758:Viral Core Proteins; D001616:beta-Galactosidase