Subnuclear localization of the trans-activating protein of human T-cell leukemia virus type I.

Human T-cell leukemia virus type I is associated with human lymphoid malignancies. The p40xI protein encoded by the x gene of this virus is believed to play some role in virally mediated transformation. This gene is known to encode a transcriptional trans activator which previous studies have shown to be a nuclear protein. Further characterization of the intracellular kinetics of this protein showed that it migrated into the nucleus very soon after synthesis. Within the nucleus, p40xI was distributed almost equally between the nucleoplasm and the nuclear matrix. Given the proposed role of the nuclear matrix in RNA transcription, the association of p40xI with the matrix places it in an appropriate cellular compartment to exercise an effect on transcription.

[Malignant lymphoma in rabbits induced by the administration of herpes virus-containing material from brown macaques]. / Zlokachestvennye limfomy u krolikov, vyzvannye vvedeniem materiala, soderzhashchego virus gerpesa burykh makakov.

Oncogenicity for rabbits of lymphotropic herpesvirus of M. arctoides (HVMA) isolated by us earlier from lymphoid cells of lines MAL-1-3 has been shown in the course of studies. Inoculation of animals both with HVMA-containing cells and cell-free virus caused the development of generalized malignant lymphomas of prolymphocytic-lymphoblastic character leading the animals to death. The molecular hybridization method permitted revealing DNA sequences related to the DNAs of B-lymphotropic herpesviruses of primates in the cells of HVMA-containing cultures, primarily-induced tumours, and in a cell line established from rabbit tumour. This fact is indicative of etiological role of HVMA in the development of malignant lymphomas in rabbits. The question about the origin of the C type particles found in MAL-1-3 and OK-1 cells remains open.

Purification of envelope glycoproteins of human T cell lymphotropic virus type I (HTLV-I) by affinity chromatography.

The external envelope glycoprotein (gp46) and transmembrane glycoprotein (gp21) of human T-cell lymphotropic virus type I (HTLV-I) were isolated from lysates of HTLV-I-infected HUT-102 cells by affinity chromatography. Fifty ml aliquots of packed HUT-102 cells were extracted with 1% Triton X-100, and lysates were treated sequentially with an affinity column containing IgG from an HTLV-I+ human subject followed by chromatography of the bound fraction over a lentil lectin column. The identity of the purified envelope proteins was confirmed with a human monoclonal antibody (0.5 alpha) to gp46 and with rabbit antisera raised to a synthetic peptide from the C-terminus of gp21. Affinity-purified envelope glycoproteins were bound to microtiter wells and used in radioimmunoassay to detect murine and human anti-envelope antibodies to gp46 and gp21 molecules.

A unique T-cell line derived from an HTLV-1-negative adult T-cell leukemia patient.

A unique T-cell line, designated ATL-5T, was established from lymphoma cells in pericardial effusion of an adult T-cell leukemia (ATL) patient not carrying HTLV-1 provirus. The cell line is OKT4 and/or Leu3a+ and OKT8 and/or Leu2a+, but interleukin 2 receptor (IL2R)- and HTLV-1 provirus genome negative, and has cytogenetically abnormal karyotypes. The cell line contains rearranged T-cell receptor beta-chain gene, which was identical in rearrangement pattern to the T-cell receptor beta-chain gene in primary cells. These results suggest that factors other than HTLV-1 may sometimes be associated with HTLV-1-negative ATL. The ATL-5T cell line we describe here is unique, and should contribute to further elucidation of the mechanisms involved in the pathogenesis of HTLV-1-negative ATL and HTLV-1-positive ATL.

IgA multiple myeloma coexistent with atypical adult T-cell leukemia.

We report a 65-yr-old male with adult T-cell leukemia (ATL) who developed multiple myeloma (MM) concomitantly. Skin lesions and peripheral leukocytosis were noted during the 5-yr observation period. There was abnormal lymphocytosis with indented or lobulated nuclei in the peripheral blood and in the bone marrow. The neoplastic cells reacted with monoclonal antibodies, OKT3, OKT4, OKIa1 and anti-Tac. His serum was positive for the antibodies to ATL-associated antigens. Human T-cell leukemia virus (HTLV) proviral DNA was detected in the leukemic cells. Thus, a diagnosis of ATL was made. There was IgA (k) paraprotein in his serum, and Bence-Jones protein (k) in urine samples. Fine needle aspiration revealed pathologic flaming plasma cells. Multiple osteolytic lesions appeared on his skull 5 yr after the initial examination. Thus, a diagnosis of MM concomitant with ATL was made.

Chromosome aberrations and clinical features of adult T cell leukemia-lymphoma not associated with human T cell leukemia virus type I.

Chromosome aberrations and clinical features of three patients with adult T cell leukemia-lymphoma (ATL) not associated with human T cell leukemia virus type I (HTLV-I) are described. From their clinical features, two patients were diagnosed as acute type and one patient was diagnosed as chronic type, which later converted to acute crisis. Clonal and many chromosomal abnormalities were observed before therapy in the two acute type cases and at relapse in the chronic type case. Karyotype aberrations, including trisomy 3, trisomy 7, trisomy 21, del(6)(q21), del(10)(p13), 14q11 translocation, and loss of X chromosome, all of which are frequently found in HTLV-I associated ATL, were also seen in these cases of HTLV-I-negative ATL.

Immunoelectron microscopic localization of the pX gene products in human T-cell leukemia virus type 1-producing cells.

The location of the pX gene products in human T-cell leukemia virus type 1-producing cells, MT-2 and HUT 102, was studied by immunoelectron microscopy using the direct and indirect peroxidase-labeled antibody methods. Fab'-peroxidase conjugates were prepared for the direct method with a maleimide compound from antisera to the carboxy-terminal region of the pX gene products. Positive immunostaining in MT-2 cells was detected in the endoplasmic reticulum, the outer and inner leaflets of the nuclear membrane, and inside their cisternae, but not in the plasma membrane and viral particles. Staining in the nucleus was faint. On the other hand, positive immunostaining in HUT 102 cells was detected diffusely in the euchromatin regions of the nucleus but not in the nucleoli, nuclear envelope, and cellular membrane systems. The location of the positive immunostaining in the HUT 102 nuclei was reconfirmed by the reaction in isolated nuclei. On the basis of both the immunoelectron microscopic and immunoblotting analyses of the pX gene products, it is suggested that the Mr 40,000 to 42,000 protein (p40x) is localized mainly in the euchromatin regions of the nuclei of human T-cell leukemia virus type 1-producing cells, and the Mr 68,000 protein (p68x) is localized mainly in the nuclear envelope and the endoplasmic reticulum of MT-2 cells. p68x detected in MT-2 cells with the anti-p40x serum was deduced to be a protein consisting of p40x and a part of env gene products and to share epitopes in common with p40x.

Anticorpos para HTLV-III por Elisa (Abbott) em pacientes hemodialisados e transplantados renais / HTLV-III antibodies by Elise (Abbott) in hemodialysis patients and renal transplantation recipients

Foram estudados 103 pacientes em hemodiálise e 94 pacientes transplantados. Nos hemodialisados, 17(16,5%) dos soros foram positivos pelo critério de duas vezes o cut-off, assim como 13(13,8%) dos soros dos pacientes transplantados. Destes 30 (15,2%) pacientes com sorologia positiva, 18(9,1%) apresentaram altos títulos (acima de cinco vezes o cut-off). Sugere-se incluir os hemodialisados e transplantados como potencialmente pertencentes a grupo de risco para infecçäo pelo HTLV-III em nosso meio e consideram ser a transfusäo sanguínea/uso de hemoderivados o provável veículo de transmissäo, tendo em vista a correlaçäo existente entre número de transfusöes e presença de sorologia positiva

A case of CD4+/CD8- adult T-cell leukemia with good response to interferon-beta terminating as a CD4+/CD8+ adult T-cell lymphoma.

The leukemic cells of adult T-cell leukemia (ATL) usually express the helper/inducer associated antigen reactive with anti-CD4 antibodies but not with anti-CD8. We present a 63-yr-old woman with ATL characterized by circulating leukemic cells with CD4+/CD8- phenotype, hepatosplenomegaly with no lymphadenopathy, and the presence of proviral DNA of human T-cell leukemia virus I in the leukemic cells. She was successfully treated with interferon beta and the remission lasted for 12 months. She then relapsed in the lymph nodes with minimal peripheral blood involvement. The neoplastic cells of the lymph node now co-expressed CD4 and CD8 antigens indicating that the change in clinical manifestation was accompanied by a phenotypic change of the leukemic cells.

Human adult T-cell leukaemia virus (ATLV) genome codes for the protein pX27 structurally homologous to immunoglobulin-like part of HLA-DQ antigen.

Within the 3' end of nucleotide sequence of human adult T-cell leukaemia virus (ATLV) four small open frames in various phases were found previously. The frames code for 10,000-, 11,000-, 12,000- and 27,000-dalton polypeptides, one of which can be responsible for virus transforming activity. By means of methods of prediction of secondary structure from amino acid composition and sequence, the beta-pleated structure was shown to be the main conformation of the 27,000-dalton polypeptide (pX27 ATLV). As a result of fitting of beta-pleats and cysteines in the N- and C-terminal parts of pX27 ATLV to those of C gamma 2 and C gamma 3 domains of human immunoglobulins G (HuIgG) and of immunoglobulin-like domains of human major histocompatibility antigens (HLA-B, -DR, -DQ), a statistically valid homology was observed by comparing the C-terminal part of pX27 ATLV with the immunoglobulin-like domain of HLA-DQw1 antigen.

A new human retrovirus isolate of West African origin (SBL-6669) and its relationship to HTLV-IV, LAV-II, and HTLV-IIIB.

A new human retrovirus of West African origin (SBL-6669) has been isolated from a patient with immunological and clinical signs of immunodeficiency. Using radioimmunoprecipitation assays (RIPA) and Western blot (WB) tests with human sera, the new virus isolate has been compared with HTLV-IV, LAV-II, and the HTLV-IIIB prototype strain of the human immunodeficiency virus (HIV). The West African isolates appeared to be members of the same virus group since their glycoproteins were antigenically indistinguishable. West African sera showed no detectable cross reaction with HTLV-IIIB glycoproteins. The external glycoprotein in the different virus strains only showed minor variations in size. The size of the transmembranous protein was not unambiguously defined. In the West African virus isolates a 30-35 kD protein was seen similar to the protein previously described possibly to represent this component. However, in SBL-6669 a distinct 41 kD protein was also identified. There were interstrain variations in the size of several viral proteins among the West African virus isolates. Only minor differences were seen between SBL-6669 and LAV-II. The variations were most pronounced in two core proteins corresponding to the 19 kD and 24 kD proteins of HTLV-IIIB. In addition, West African human retroviruses appear to differ in pathogenicity. LAV-II and SBL-6669 are associated with immunodeficiency, whereas HTLV-IV was isolated from healthy individuals. Since further spread of these viruses to other parts of the world is imminent, it is necessary to consider their antigenic and immunogenic properties in serodiagnosis of HIV infections and in planning for immunoprophylactic interventions.

Isolation of a lentivirus from a macaque with lymphoma: comparison with HTLV-III/LAV and other lentiviruses.

A retrovirus has been isolated on the human T-cell line HuT 78 after cocultivation of a lymph node from a pig-tailed macaque (Macaca nemestrina) that had died with malignant lymphoma in 1982 at the University of Washington primate center. This isolate, designated MnIV (WPRC-1) (M. nemestrina immunodeficiency virus, Washington Primate Research Center) shows the characteristic morphology of a lentivirus and replicates to high titers in various lymphocyte lines of human and primate origin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified MnIV revealed multiple bands of structural proteins, including a major viral gag protein of 28 kilodaltons, that did not comigrate with the viral proteins of a human immunodeficiency virus (HIV [FRE-1]) that was also isolated on HuT 78 cells. The relatedness of MnIV to other lentiviruses (HTLV-III/LAV, EIAV, and visna) was examined in radioimmunoassays, by immunoblot techniques, and by N-terminal amino acid sequence analysis of the viral p28 gag protein. The immunoassays revealed cross-reactivity only between MnIV p28 and HTLV-III/LAV p24, and sequence analysis showed that 14 of the 24 N-terminal residues of MnIV p28 and HTLV-III/LAV p24 are identical. These results indicate that MnIV belongs to the same lentivirus family as HTLV-III/LAV but is only partially related to these human acquired immune deficiency syndrome retroviruses.

Detection of AIDS virus in macrophages in brain tissue from AIDS patients with encephalopathy.

One of the common neurological complications in patients with the acquired immune deficiency syndrome (AIDS) is a subacute encephalopathy with progressive dementia. By using the techniques of cocultivation for virus isolation, in situ hybridization, immunocytochemistry, and transmission electron microscopy, the identity of an important cell type that supports replication of the AIDS retrovirus in brain tissue was determined in two affected individuals. These cells were mononucleated and multinucleated macrophages that actively synthesized viral RNA and produced progeny virions in the brains of the patients. Infected brain macrophages may serve as a reservoir for virus and as a vehicle for viral dissemination in the infected host.

Human T-cell leukemia virus I provirus and antibodies in a captive gorilla with non-Hodgkin's lymphoma.

Antibodies reactive against human T-cell leukemia virus I (HTLV-I) were detected by indirect immunofluorescence assay using MT-2 as target cells, enzyme linked immunosorbent assay screen and competition assay, and Western blot analysis in three sera (one collected in 1979) from a captive gorilla which developed diffuse histiocytic lymphoma in 1983. The sera from four other healthy gorillas housed separately were HTLV-I antibody negative. All sera were negative for HTLV-III antibodies by enzyme linked immunosorbent assay. Southern blot analysis of DNA from lymphoma tissue after digestion with BamHI and using complete HTLV-I genome probe gave one 10-kilobase fragment and a characteristic 1.05-kilobase internal fragment detected in all known HTLV-I isolates. These results indicate that the gorilla was infected with HTLV-I or a closely related simian virus several years before the development of lymphoma.

Interleukin 1 alpha mRNA in virus-transformed T and B cells.

IL-1 alpha cDNA clone was isolated from a T cell line infected by the human T lymphotropic retrovirus type-I (HTLV-I/ATLV). We found significant amounts of mRNA hybridizing to IL-1 alpha cDNA not only in HTLV-I-transformed T cells but also in Epstein-Barr Virus-transformed B cells. A part of IL-2 receptor inducing activity in Adult T cell leukemia (ATL) cell line seems to be due to IL-1 alpha.

Biological and biochemical characterization of a cloned Leu-3- cell surviving infection with the acquired immune deficiency syndrome retrovirus.

Leu-3- cells that survive infection with the acquired immune deficiency syndrome (AIDS) retrovirus can be induced with IUdR to express infectious virus. A cellular clone (8E5), isolated by limiting dilution of a mass culture of survivor cells, was found to contain a single, integrated provirus that was constitutively expressed. Although IUdR treatment of 8E5 cells failed to induce infectious virus, cocultivation with Leu-3+ cells generated the characteristic syncytia associated with acute AIDS retrovirus infection. The single integrated copy of proviral DNA directs the synthesis of all major viral structural proteins except p64, as monitored by immunoblotting. The relationship of the 8E5 clone to viral latency and persistence is discussed.