Identification of HTLV-I- or HTLV-II-producing cells by cocultivation with BHK-21 cells stably transfected with a LTR-lacZ gene construct.
The Syrian Hamster kidney cell line (BHK-21) was stably transfected with a plasmid vector containing the lacZ bacterial gene under the control of a HTLV-I-LTR promoter. In these cells termed pA18G-BHK-21, this lacZ construct is inducible by the tax protein produced by a tax expression vector. It was also shown that beta-galactosidase synthesis was detected within 48 h after cocultivation of pA18G-BHK-21 cells with HTLV-I (HUT-102, MT2, C91/PL, 2060) or HTLV-II (MoT strain) -producing cells. The number of positive cells was directly related to the number of HTLV-I or -II-infected cells seeded. In addition, the LTR transactivation observed in coculture with HTLV-I-infected cells was specifically inhibited by sera containing antibodies directed against HTLV-I proteins, but not, or only weakly, by sera containing HTLV-II antibodies. Conversely, beta-galactosidase expression induced by HTLV-II-infected cells was inhibited by sera of HTLV-II-infected individuals, but not, or only weakly, by HTLV-I-positive sera. Irrespective of the inducer cell, sera from uninfected people did not inhibit LTR-driven expression of the lacZ gene in pA18G-BHK-21 cells cocultivated with HTLV-producing cells. This assay may thus be employed profitably for the detection and quantification of both HTLV-producing cells and HTLV-specific antibodies.
Astier-Gin
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INSERM Unité 328, Bordeaux, France.
Portail
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JP
Lafond
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F
Guillemain
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B
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D005822:Genetic Vectors; D015481:HTLV-I Antibodies / Q000097:blood; D015482:HTLV-II Antibodies / Q000097:blood; D015368:Human T-lymphotropic virus 1 / Q000235:genetics* / Q000502:physiology*; D015367:Human T-lymphotropic virus 2 / Q000502:physiology*; D006801:Humans; D007763:Lac Operon; D009500:Neutralization Tests; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D015533:Transcriptional Activation; D014162:Transfection; D014773:Virology / Q000379:methods; D014779:Virus Replication; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D015481:HTLV-I Antibodies; D015482:HTLV-II Antibodies; D001616:beta-Galactosidase