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Biomedical subjects

K Shimotohno

Publications and source records attributed to K Shimotohno.

At least 199 records · Page 11Linked to original sources

Augmentation of human immunodeficiency virus type 1 gene expression by tumor necrosis factor alpha.

It is now well established that cytokines are involved in the regulation of gene expression from HIV-1 LTR. The present study provides evidence that TNF-alpha stimulates HIV-1 gene expression and that the enhancer sequence within the HIV-1 LTR is involved in the stimulation. These results support the idea that immunologic stimulation and infection may trigger the development of clinical AIDS in individuals latently infected with HIV-1.

Animals↗

Multi-step carcinogenesis model for adult T-cell leukemia.

The age-specific occurrence of adult T-cell leukemia (ATL) was analyzed using 357 cases collected during nationwide surveys carried out between 1982 and 1985 in Japan. A simple Weibull distribution function fitted well as a model. The mode of ATL onset was log-linear in this model and the curves for males and females overlapped completely. The presence of age-dependent accumulation of leukemogenic events within human T-cell leukemia virus type 1-immortalized T cells was suggested prior to the development of ATL, and the approximate number of independent leukemogenic events in ATL is estimated to be five. This stochastic analysis supported a multi-step carcinogenesis as an appropriate model for ATL.

Adult↗

DNA blotting analysis of human retroviruses in cerebrospinal fluid of spastic paraparesis patients: the viruses are identical to human T-cell leukemia virus type-1 (HTLV-1).

The structure of the integrated provirus in cell lines established from the cerebrospinal fluid (CSF) of patients with human T-cell leukemia virus type-1 (HTLV-1)-associated myelopathy (HAM) was analyzed. The digestion patterns with 3 restriction endonucleases, Sac I, Eco RI and PstI, of the proviruses integrated in T-lymphoid cell lines derived from the CSF of 4 HAM patients were similar to those of HTLV-1 from adult T-cell leukemia (ATL) patients. Integrated proviruses in one of these cell lines derived from the CSF were further analyzed in detail with 2 more restriction enzymes, BamHI and Sma I. The results indicate that the retrovirus found in lymphocytes of the CSF from patients with HAM are very similar, if not identical, to HTLV-1 found in the leukemic lymphocytes of ATL patients.

Cell Line↗

Production of parathyroid hormone-related protein in adult T-cell leukemia cells.

Human parathyroid hormone-related protein (PTHrP) mRNA was detected in peripheral leukemic cells obtained from adult T-cell leukemia (ATL) patients as well as in cultured human T-cell leukemia virus type I (HTLV-I)-infected T-cell lines. In contrast, PTHrP mRNA was not detected in other types of leukemic cells. Using radioimmunoassay, immunoreactive PTHrP was also detected in the spent media of HTLV-I-infected T-cell lines. These results suggest that PTHrP plays an important role in developing the hypercalcemia frequently observed in ATL patients.

HTLV-I Infections↗

Identification of a cis-regulatory element involved in accumulation of human T-cell leukemia virus type II genomic mRNA.

The X gene products of the human T-cell leukemia viruses type I and II are thought to be involved not only in viral replication but also in mediating the expression of certain cellular genes. These X gene products are known to be translated from doubly spliced viral mRNA, while viral structural proteins, such as the gag, pol, and env gene products, are translated from unspliced or singly spliced viral mRNA. One of the X gene products of human T-cell leukemia virus type II, tax2 protein, has been shown to be responsible for transcriptional stimulation from the viral long terminal repeat. The other X gene product(s) of human T-cell leukemia virus type II, the rex2 protein(s), is located in the nuclear fraction of virus-infected cells, but its function is not known. This article reports evidence that rex2 protein(s) enhances the accumulation of unspliced viral RNA by interacting posttranscriptionally, either directly or indirectly, with a cis-regulatory element downstream from the first splice donor site in the long terminal repeat.

Cell Line↗

Nucleotide sequence analysis of a provirus derived from HTLV-1-associated myelopathy (HAM).

Human T-cell leukemia virus type 1 (HTLV-1) is known to be associated with adult T-cell leukemia (ATL). Recently, HTLV-1-associated myelopathy (HAM) was described as a neurological disease with which an etiological association of HTLV-1 is suspected. A provirus genome was cloned from a lymphoid cell line derived from the cerebrospinal fluid of a patient with HAM, in order to examine in detail the etiological virus associated with HAM. The nucleotide sequence of the long terminal repeat (LTR), protease, env and pX regions of the provirus shows over 97% homology with that of HTLV-1 derived from ATL. These results suggest that this provirus, derived from a patient with HAM, belongs to the same species as HTLV-1 derived from patients with ATL.

Amino Acid Sequence↗

Evidence for phosphorylation of trans-activator p40x of human T-cell leukemia virus type I produced in insect cells with a baculovirus expression vector.

Human T-cell leukemia virus type I (HTLV-I) encodes a trans-activator protein p40x which positively regulates transcription of the viral RNA as well as interleukin-2 and its receptor genes. We placed a cDNA coding for p40x in baculovirus Bombyx mori nuclear polyhedrosis virus (BmNPV) expression vectors. Infection of BmN cells derived from an insect, B. mori (silkworm), with a recombinant virus led to the production of soluble p40x. The biological activity of the recombinant p40x was demonstrated by introducing the protein into intact NIH 3T3 cells that had been selected for genomic integration of HTLV-I LTR connected with the CAT gene. Immunocytochemical and cell fractionation analyses showed the localization of p40x in both the cytoplasmic and nuclear fractions of BmN cells. Analyses of 32P-labeled proteins of BmN cells by cell fractionation and subsequent immunoprecipitation revealed that the p40x present in each subcellular fraction was phosphorylated. The post-translational modification was inhibited by the addition of a protein kinase inhibitor K252a during the metabolic labeling of BmN cells. Phosphoamino acid analysis indicated that the phosphorylation occurred on serine residues of p40x.

Animals↗

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.

Cell Line↗

U3 sequences from HTLV-I and -II LTRs confer pX protein response to a murine leukemia virus LTR.

Human T-cell leukemia virus (HTLV) types I and II are unusual among replication-competent retroviruses in that they contain a fourth gene (chi) necessary for replication. The chi gene product, p chi, transcriptionally transactivates the viral long repeat (LTR), and is thus a positive regulator. To investigate p chi transactivation, sequences from the U3 regions of the LTRs of HTLV-I and -II were inserted into the Moloney murine leukemia virus (M-MuLV) LTR by recombinant DNA techniques. Transient expression assays of the chimeric LTRs indicated that the HTLV sequences conferred to the M-MuLV LTR responsiveness to HTLV p chi protein. M-MuLV enhancers were not required for function of the chimeric LTRs. Infectious recombinant M-MuLVs containing chimeric LTRs were also generated. These viruses showed higher infectivity when assayed in mouse cells expressing HTLV-II p chi protein compared to normal mouse cells. Thus the HTLV sequences were able to confer p chi responsiveness to infectious M-MuLV. The generation of a virus dependent on a transactivating protein for its replication has implications for the evolution of the human T-cell leukemia viruses.

DNA, Viral↗

Total nucleotide sequences of the infectious cloned DNAs of bean golden mosaic virus.

Complete nucleotide sequences of the infectious cloned DNA components (DNA A and B) of bean bolden mosaic virus were determined. The DNA A (2585 nucleotides) and DNA B (2647 nucleotides) have little sequence homology with each other, but both A and B contain a common region of 205 nucleotides. A possible large hairpin structure is detected in the common region. Nucleotide sequences of DNAs A and B revealed the presence of 8 potential coding regions for proteins (m.w. greater than 10,000). Among them, four open reading frames (ORFs 1-4) encode proteins of m.w. 30,000 or greater, and are individually coded in virion DNA A and B senses (+) and their complementary senses (-), respectively. The other four ORFs 5-8 are in virion DNA B(+) and its complementary sense (-). All of the ORFs 1-4 have regulatory signals for RNA synthesis (TATAA/T) in the region 5' upstream from a potential start codon ATG.

Base Sequence↗

Identification of p26Xb and p24Xb of human T-cell leukemia virus type II.

Human T-cell leukemia virus type II (HTLV-II) isolated from a T-cell variant of hairy cell leukemia contains gag, pol and env genes as well as a fourth gene termed X, which can code three major open reading frames Xa, Xb and Xc. Proteins with molecular masses of 26 kDa (p26Xb) and 24 kDa (p24Xb) encoded by the Xb open reading frame were identified with antisera directed against synthetic peptides corresponding to the N-terminal and C-terminal amino acid sequences deduced from the structure of the Xb open reading frame. More than half the Xb products were found to be located in the nuclear fraction of HTLV-II-infected cells.

Antigen-Antibody Complex↗

A sequence homology between the pX genes of HTLV-I/II and the murine IL-3 gene.

Searching the protein sequence database for amino acid sequences homologous to the x-lor sequence in the pX region of human T-cell leukemia virus types I and II (HTLV-I/II), we found that there is a region of 38 amino acids where the murine interleukin 3 (IL-3) sequence has a 40% homology with the x-lor sequence. A statistical analysis shows that this homology is highly significant with a probability of 1.57 X 10(-10). The biological implication of this homology is discussed.

Amino Acid Sequence↗

Molecular cloning of unintegrated closed circular DNA of porcine retrovirus.

Viral DNA of unintegrated closed circular form was isolated from a swine kidney cell line (SKL) which was infected with a porcine retrovirus Tsukuba-1 (PRetV) produced from a swine malignant lymphoma-derived cell line. Shimozuma-1 and cloned using a lambda phage vector, Charon 21A. One of ten independent clones contained the 8.3 kb DNA fragment as an insert, which was thought to be a full length of viral DNA molecule carrying a long terminal repeat (LTR) sequence. We have analyzed this insert by mapping the recognition sites of some restriction endonucleases by Southern blot hybridization with appropriate probes.

Animals↗

Detection of the X gene product of simian T-cell leukemia virus.

The gene product of the X region was examined in simian lymphoid cell lines producing simian T-cell leukemia virus (STLV), which is closely related to human T-cell leukemia virus (HTLV). By use of specific antibodies against pX peptides of HTLV-I, a protein of 41 kDa was identified as a pX product of STLV.

Animals↗

Adult T-cell leukemia/lymphoma not associated with human T-cell leukemia virus type I.

We describe five patients with adult T-cell leukemia/lymphoma (ATL) with neither integration of human T-cell leukemia virus type I (HTLV-I) into their leukemia cells nor anti-HTLV-I antibody in their sera. These findings indicate that HTLV-I may not have been involved in leukemogenesis in these patients. The clinicohematological, cytopathological, and immunological features of HTLV-I-negative ATL were exactly the same as those of HTLV-I-associated ATL. Leukemia cells with pleomorphic nuclei, generalized lymphadenopathy, hepatosplenomegaly, skin lesions, hypercalcemia, and elevated lactate dehydrogenase levels, all of which are characteristic features of typical ATL, were also seen in these patients with HTLV-I-negative ATL. Leukemia cells expressed T3, T4, and pan-T-cell antigens in three cases, and T3 and pan-T-cell antigens in two. All five patients had lived in ATL-nonendemic areas. The finding of HTLV-I-negative ATL suggests that factor(s) other than HTLV-I infection may be involved in ATL leukemogenesis.

Adult↗