Identification of HTLV-I- or HTLV-II-producing cells by cocultivation with BHK-21 cells stably transfected with a LTR-lacZ gene construct

Identification of HTLV-I- or HTLV-II-producing cells by cocultivation with BHK-21 cells stably... 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 β-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, β-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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Virological Methods Elsevier

Identification of HTLV-I- or HTLV-II-producing cells by cocultivation with BHK-21 cells stably transfected with a LTR-lacZ gene construct

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Publisher
Elsevier
Copyright
Copyright © 1995 Elsevier Ltd
ISSN
0166-0934
eISSN
1879-0984
D.O.I.
10.1016/0166-0934(94)00097-Z
Publisher site
See Article on Publisher Site

Abstract

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 β-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, β-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.

Journal

Journal of Virological MethodsElsevier

Published: Jan 1, 1995

References

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