Biomedical subjects

Y Aono

Publications and source records attributed to Y Aono.

28 records · Page 2Linked to original sources

PubMed1992

Rapid, sensitive, specific, and quantitative detection of human T-cell leukemia virus type 1 sequence in peripheral blood mononuclear cells by an improved polymerase chain reaction method with nested primers.

Improving on the nested double polymerase chain reaction (PCR) described previously, we have developed a new two-step PCR (TS-PCR) method for detecting more specifically the human T-cell leukemia virus type 1 (HTLV-1) proviral sequences in peripheral blood mononuclear cells (PBMC). In our TS-PCR method, the point of modification is to use optimal concentrations of primers in the first amplification step in the range of 0.01-0.025 microM. This increases sensitivity and specificity enough to detect from 1 to 10(5) copies of template DNA without radioisotopes. This method is rapid because of completion in 1 day and is also applicable for quantitative detection of clinical specimens. The data show that the quantitative detection of HTLV-1 proviral sequences by this method correlates with the anti-HTLV-1 antibody titers from serologic analysis of seropositive healthy carriers. Moreover, the TS-PCR method using each specific primer was also attempted for successful detection of other viral genomes; therefore, the principle of this method is widely suitable for routine detection of genomes in the basic and clinical microbiological fields.

Base Sequence↗

PubMed1992

Target epitope in the Tax protein of human T-cell leukemia virus type I recognized by class I major histocompatibility complex-restricted cytotoxic T cells.

A trans-acting regulatory gene product p40tax (Tax) of human T-cell leukemia virus type I (HTLV-I) is one of the main target antigens recognized by cytotoxic T lymphocytes (CTL) specific for HTLV-I. A CTL epitope within the Tax protein was identified in this report. HTLV-I-specific CD8+ CTL lines established from two HTLV-I carriers with HTLV-I-associated myelopathy or Sjögren syndrome were previously demonstrated to kill predominantly the target cells expressing HTLV-I Tax. The CTL from two patients showed significant levels of cytotoxicity to autologous target cells pulsed with a synthetic peptide of 24 amino acids corresponding to the amino-terminal sequences of the Tax protein. Allogeneic target cells were also sensitized for CTL by this peptide when the target cells have HLA-A2. Tax-specific cytotoxicity, detected as cytolysis of the target cells infected with vaccinia virus-HTLV-I recombinant expressing Tax protein, was almost completely inhibited by competitor cells pulsed with the synthetic peptide. This indicates that a major CTL epitope is present in this peptide. Further analysis using shorter peptides revealed that the core sequence of the CTL epitope was LLFGYPVYV at positions 11 through 19. This sequence can be aligned with the HLA-A2-specific motifs reported recently.

Amino Acid Sequence↗

PubMed1991

A novel alternatively spliced viral mRNA transcribed in cells infected with human T cell leukemia virus type 1 is mainly responsible for expressing p21X protein.

The pX sequence of human T cell leukemia virus type 1 (HTLV-1) has been thought to be expressed as a doubly spliced mRNA that codes for p40tax, p27rex and p21X. However, we identified a novel alternatively spliced mRNA in the HTLV-1 infected cells by using reverse transcription followed by the polymerase chain reaction. This mRNA contains only the first and third exons of the doubly spliced mRNA and encodes only p21X. Our data that this mRNA is responsible for expressing p21X exists in most of HTLV-1 infected cells strongly suggests that p21X may play a crucial role for HTLV-1 replication.

Base Sequence↗

PubMed1991

[Measurement of anti-microsomal and anti-thyroglobulin antibodies by radioimmunoassay].

In this study, we measured anti-microsomal (same as anti-thyroid peroxidase) and anti-thyroglobulin antibodies (anti-TPOAb and anti-TgAb) in sera by new RIA kits and compared with the conventional particle agglutination (PA) kits. Two of 80 normals (2.5%) showed anti-TPOAb positive activity (greater than or equal to 0.3 unit/ml), and 9 of same samples (11.3%) showed anti TgAb positive activity (greater than or equal to 0.3 unit/ml). Sixty of 67 patients with Graves' disease (89.6%), and 14 of 22 with Hashimoto's disease (63.6%) were anti-TPOAb positive, and 42 (62.7%), and 18 (81.8%) were anti-TgAb positive respectively. Samples of 5 non-autoimmune disease were all negative by TPOAb and TgAb assays, suggesting the high specificity of new RIA kits to the autoimmune thyroid diseases. The RIA and PA methods showed a good correlation (anti-TPOAb:rs = 0.9134, anti-TgAb:rs = 0.7016). However, several negative sera in PA assay showed positive values in the RIA, so it was showed that new RIA kits were more sensitive than the PA assays.

Autoantibodies↗

PubMed1989

Accelerated passive Heymann nephritis in rats as an experimental model for membranous glomerulonephritis and effects of azathioprine and prednisolone on the nephritis.

Progressive passive Heymann nephritis (group II) was induced in rats by i.v. injections of rabbit antiserum against an antigen from the brush border of the proximal tubules of rat kidney following immunization with rabbit gamma-globulin in Freund's complete adjuvant, and the process of the disease was compared with that of rats that received the antiserum alone (group I). The rats of group I showed proteinuria (30-50 mg/day) and plasma cholesterol content (80-90 mg/dl) slightly higher than the normal level from the 17th or the 22nd day after antiserum injection to the 90th day. The rats of group II revealed a heavy proteinuria (300-400 mg/day) and hypercholesterolemia (approx. 200 mg/dl) during the same period. In group II, there were the thickening of glomerular basement membrane (GBM) and spike formation. Moreover, granular deposits of rat IgG, rabbit IgG and rat C3 were observed along the GBM. These changes were weaker in group I. When given orally, daily, azathioprine (20 mg/kg) and prednisolone (1 and 3 mg/kg) showed a beneficial effect on the nephritis in group II. The group II model closely resembles human glomerulonephropathy and may be useful for studying the effect of medication on glomerulonephropathy.

Animals↗

PubMed1971