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  • Title: Simple assay system for detecting human T cell leukemia virus type I-binding cells and its application in titrating binding inhibitory antibodies.
    Author: Akari H, Kannagi M, Shinjo T, Harada S.
    Journal: Lab Invest; 1993 Nov; 69(5):629-34. PubMed ID: 8246453.
    Abstract:
    BACKGROUND: A simple and rapid assay system for the detection of human T cell leukemia virus type I (HTLV-I) binding cells was developed to assess the virus specific receptor and titrate the antibodies to block the virus binding. EXPERIMENTAL DESIGN: Cells (5 x 10(5)) were incubated with 100 microliters of the concentrated HTLV-I at 37 degrees C for 1 hour. After washing, the cells were reacted with anti-HTLV-I envelope monoclonal antibody (rat) for 30 minutes on ice and then stained with fluorescein-isothiocyanate-conjugated anti-rat immunoglobulin. The stained samples were analyzed on FACScan. Antibody-titration of the virus-binding inhibition was carried out by pretreatment of the virus with serially diluted sera. RESULTS: The specificity of the virus-binding was shown by dose-response relationship, kinetics of the binding, and temperature dependency. HTLV-I was absorbed onto a wide range of human cell lines and peripheral blood lymphocytes at various levels. Antibodies to inhibit the virus-binding were also quantitatively detected in sera from HTLV-I infected individuals, including asymptomatic carriers and patients with adult T cell leukemia or HTLV-I-associated myelopathy, but not from healthy seronegative controls. CONCLUSIONS: This assay system would be useful in screening the virus-specific receptor and the neutralizing antibodies to HTLV-I. Thus, the assay could be applied to further studies on HTLV-I-related diseases.
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