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

J Virol Methods. 1995 Jan;51(1):19-29. doi: 10.1016/0166-0934(94)00097-z.

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 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cricetinae
  • Genetic Vectors
  • HTLV-I Antibodies / blood
  • HTLV-II Antibodies / blood
  • Human T-lymphotropic virus 1 / genetics*
  • Human T-lymphotropic virus 1 / physiology*
  • Human T-lymphotropic virus 2 / physiology*
  • Humans
  • Lac Operon
  • Neutralization Tests
  • Plasmids / genetics
  • Promoter Regions, Genetic
  • Repetitive Sequences, Nucleic Acid
  • Transcriptional Activation
  • Transfection
  • Virology / methods
  • Virus Replication
  • beta-Galactosidase / biosynthesis
  • beta-Galactosidase / genetics

Substances

  • HTLV-I Antibodies
  • HTLV-II Antibodies
  • beta-Galactosidase