ABSTRACT
A quantitative real-time PCR assay was developed to measure the proviral load of human T-lymphotropic virus type I (HTLV-I) in peripheral blood. The technology utilizes special primers and Taqman MGB fluorescence probe to measure amplification products from the gag-pro-pol polyprotein gene of HTLV-I. HTLV-I copy number was normalized to the amount of cellular DNA by quantitation of the beta-actin gene, The amplification system was sensitive to detect 5 copy/microL. The standard curve had a good linearity when the quantity for the gene was between 10(3) and 10(7) copy/microL (R2 = 0.999). Good reproducibility was observed in each intra- and inter-assay. We also measured proviral load in peripheral blood in 12 HTLV-I seropositive former blood donors. Proviral load for HTLV-I infected donors ranged from 0.015 to 12.819 copy/cell in WBC with the mean of 3.116 copy/cell.
Subject(s)
Humans , Gene Products, gag , Genetics , Gene Products, pol , Genetics , Human T-lymphotropic virus 1 , Genetics , Molecular Probes , Polymerase Chain Reaction , Methods , Viral Proteins , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate effects of anti-dsDNA autoantibodies on growth of tumor in vitro and in vivo.</p><p><b>METHODS</b>BALB/c mice were inoculated with inactivated tumor cells and challenged s.c. with SP 2/0 and Wehi 164 tumor cells four weeks after the last inoculation. The naïve mice were inoculated with SP 2/0 tumor cells immediately after incubating with sera derived from the immunized mice at week 6. Then the tumor size was examined. In vitro, the cytotoxicity of anti-dsDNA autoantibodies to tumor cells was analysed. Furthermore, apoptosis of SP 2/0 and Wehi 164 tumor cells induced by anti-dsDNA autoantibodies was examined by FACS.</p><p><b>RESULTS</b>In vivo study showed that the growth of SP 2/0 and Wehi 164 tumors were inhibited in mice with anti-dsDNA autoantibodies, but not in mice lack of anti-dsDNA autoantibodies. In vitro, apoptosis of SP 2/0 and Wehi 164 tumor cells was induced when the tumor cells were incubated with the sera containing anti-dsDNA autoantibodies. Statistical analysis showed that the ability of anti-dsDNA autoantibodies to induce apoptosis of SP 2/0 and Wehi 164 tumor cells was significantly correlated with affinity (r = 0.990, P < 0.01 and r = 0.901, P < 0.05).</p><p><b>CONCLUSION</b>Anti-dsDNA autoantibodies have inhibitory effect on tumor cells via inducing apoptosis.</p>