Abstract Cynomolgus monkeys and squirrel monkeys were inoculated with autologous lymphoid cell lines immortalized by and producing human T‐cell leukemia virus type‐1 (HTLV‐I) in order to serve as an animal model of adult T‐cell leukemia (ATL). The autologous cell tines were established from peripheral blood mononuclear cells (PBMC) from each monkey by co‐cultivation with lethally irradiated MT‐2 cells producing HTLV‐I. All of these cell lines, which had monkey karyotypes, grew continuously without addition of interleukin‐2 (IL‐2) and expressed virus‐specific proteins of HTLV‐I and IL‐2 receptor. After inoculation with the autologous cell lines, specific antibodies against HTLV‐I proteins could be detected in their plasma, and transformed HTLV‐I‐infected cells could be recovered from their peripheral blood for at least 6 months. However, no signs of ATL have been observed to date, i.e. 2 years after inoculation.
Abstract A new mouse monoclonal antibody (HIEI, IgG1 type) that reacts with a cell surface glycoprotein of human lymphocytes was isolated. Membrane immunofluorescence assay showed that HIEI, like the anti‐Tac monoclonal antibody, reacted preferentially with activated normal human T‐cells and adult T‐cell leukemia (ATL) virus (ATLV)‐carrying human T‐ and B‐cell lines. However, an interesting difference between HIEI and anti‐Tac antibody was that HIEI did not react with ATLV‐transformed simian cell lines or those cultured with interleukin‐2 (IL‐2), whereas the anti‐Tac antibody did. The immunoprecipitation assay showed that both HIEI and anti‐Tac antibody precipitated a glycoprotein with a molecular weight of 60,000 daltons (gp60) from activated normal T‐cells and ATLV‐positive T‐ and B‐cells, and also gp53 from MT‐2 and MT‐2‐related T‐cell lines transformed with ATLV in vitro by the MT‐2 cocultivation method. HIEI inhibited the Independent proliferation of normal T‐cells, but its inhibitory effect was much weaker than that of the anti‐Tac antibody. The anti‐Tac antibody interfered with the binding of HIEI to target cells, but HIEI did not block binding of the anti‐Tac antibody to the cells. These observations indicate that HIEI antibody recognizes a new antigenic determinant of the human Tac antigen.
Abstract Two new murine monoclonal IgG1 antibodies, H‐31 and H‐A26, were characterized in comparison with two previously obtained monoclonal antibodies against human interleukin 2 (IL‐2) receptor (IL‐2 R), anti‐Tac and HIEI. In immunofluorescence assays with various human hematopoietic cells, H‐31 and H‐A26 antibodies both reacted with only IL‐2 R‐positive cells, and they precipitated IL‐2 R molecules, glycoproteins with molecular weights of 60K and 53K daltons (gp60/gp53), from human T‐cell leukemia virus type I (HTLV‐I)‐carrying MT‐2 cells, as demonstrated by sequential immunoprecipitation after absorption of IL‐2 R with anti‐Tac. Antibody‐binding competition assays showed that H‐31 and anti‐Tac, and H‐A26 and HIEI, respectively, competed reciprocally in binding to the cells, and that anti‐Tac also inhibited the binding of HIEI but not vice versa . H‐31, like anti‐Tac, strongly inhibited the IL‐2‐dependent proliferation of normal activated T‐cells, absorption of IL‐2 and direct binding of IL‐2 to the cells, while H‐A26, like HIEI, inhibited those processes only weakly. The spectra of reactivities of these antibodies with various simian cell lines derived by HTLV‐I infection were different, as revealed by immunofluorescence studies. Human IL‐2 R was shown to express a unique antigenic determinant, detected with HIEI, that was not detectable in IL‐2 R molecules of Old and New World monkeys, and also to express determinants common to simian IL‐2 R molecules. These observations indicate that H‐31 and H‐A26 recognize human IL‐2 R molecules and that the antigenic sites on the IL‐2 R molecule defined by H‐31, H‐A26, anti‐Tac, and HIEI are different.
Two glycoproteins (HN and F) of parainfluenza virus were immunogold-labeled with polyclonal and monoclonal antibodies, respectively, and their labeling patterns were compared. Both glycoproteins HN and F were efficiently and homogeneously labeled with polyclonal antibodies, whereas they were labeled much less and heterogeneously with monoclonal antibodies. When either protein was initially labeled with monoclonal antibody, and then the other one, with polyclonal antibody, immunolabels of two glycoproteins were almost completely segregated. Although this segregation deformed virion morphology, it supported the concept of monoclonal antibody-mediated movements of glycoproteins.
We showed previously that WKA (rat MHC, RT-1k) and DA (RT-1a) rat CTL specific for human T cell leukemia virus type-I (HTLV-I) recognized the gag and pX gene-encoded Ag. In the present study, we explored HTLV-I Ag recognized by CTL from other MHC genotype (RT-1l) rats, LEW and F344, and examined whether HTLV-I Ag expressed by recombinant vaccinia viruses (rVV) could prime WKA and LEW rats for HTLV-I-specific CTL. Upon priming in vivo and repetitive stimulation in vitro with HTLV-I+ syngeneic T cells, HTLV-I-specific CD8+ CTL were demonstrated in LEW and F344 rat spleen cell cultures. Interestingly, these CTL were directed against HTLV-I env and pX gene products. Immunization of LEW and WKA rats with the env and gag gene-expressing rVV, respectively, resulted in generation of HTLV-I-specific CD8+ CTL. However, a pX-gene expressing rVV failed to prime either rat strain. In addition, HTLV-I-specific CD8+ CTL from F1 hybrid (WKA x LEW) rats generated by immunization and restimulation with HTLV-I+ syngeneic T cells showed gag and pX Ag specificity on WKA rat cells and env and pX Ag specificity on LEW rat cells. These results suggest that immunogenicity of HTLV-I gag and env Ag in induction of HTLV-I-specific CTL varies depending on MHC, and that the pX Ag expressed by rVV is not a potent immunogen for rats.
We reported a rare case of spontaneous intramuscular hematoma associated with alcoholic liver cirrhosis patient. A 60-year-old man with alcoholic liver cirrhosis was admitted to our hospital for pneumonia. It was improved by conservative treatments, however, he was complaint of severe pain in right hip and thigh. MRI revealed intramuscular hematoma at acute phase. Resting and anticoagulant therapy reduced it, but liver functions were not improved. After a few months, he received another intramuscular hematoma in the left thoracic muscles, and then died because of liver failure. It seems important to consider intramuscular hematoma as a complication of alcoholic liver cirrhosis.
