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K Ikuta

Publications and source records attributed to K Ikuta.

At least 343 records · Page 19Linked to original sources
PubMed1986

A joining-diversity-joining complex generated by inversion mechanism and a variable-diversity complex in the beta-chain gene of the human T-cell receptor.

We have analysed an inactive allele of the beta-chain gene of the T-cell receptor in a human T-cell line HPB-ALL. Comparison with germline sequences showed that HPB-ALL has a joining (J)-diversity (D)-J complex recombined in head-to-head configuration and a variable (V)-D complex in tail-to-tail configuration. These results demonstrate that the inversion mechanism functions in the beta-chain gene of the T-cell receptor. The presence of the V-D complex suggests that V-D recombination could occur prior to D-J recombination although there is no definite proof that the V-D complex is an intermediate to form the V-D-J complex.

Base Sequence↗
PubMed1986

Polypeptide analysis with monoclonal antibodies of A type inclusion bodies induced by cowpox virus.

The antigen in A type inclusion bodies (ATIB) induced by cowpox virus (CPV) was examined by immunofluorescent staining with monoclonal antibodies and polypeptide analysis. In the immunofluorescence (IF) test, these monoclonal antibodies reacted only with cytoplasmic inclusion bodies in cells infected with CPV. The fluorescence became detectable in the cells 6-9 hours after infection with CPV. No fluorescence was detectable in cells infected with CPV in the presence of cytosine-I-beta-D-arabinofuranosyl-HCl (Ara C) or in cells infected with other poxviruses, such as vaccinia virus (VV) or Shope fibroma virus (SFV). On Western blotting and immunoprecipitation followed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), only one component with a molecular weight of about 160,000 (160 K) was detected in CPV-infected cells. This 160 K polypeptide was first detectable 12 hours after infection of cells with CPV, and was not detectable in infected cells in the presence of Ara C. The 160 K polypeptide was not detected in cells infected with VV or SFV, or in virions purified from CPV-, VV- or SFV-infected cells.

Animals↗
PubMed1986

Similarities and dissimilarities in the structure and expression of viral genomes of various virus strains immunologically related to Marek's disease virus.

Various strains immunologically related to Marek's disease virus (MDV) have been subdivided into three serotypes: serotype 1, pathogenic strains of MDV and attenuated or apathogenic variants derived from them; serotype 2, naturally occurring apathogenic strains of MDV; serotype 3, herpesvirus of turkey (HVT). The viral genome structures of these three serotypes were compared by a simple, practical method using total DNA extracted from virus-infected cells instead of viral DNA purified from virions. The restriction endonuclease-cleavage patterns of serotype 2 viral DNA were found to differ from those of either serotype 1 MDV or serotype 3. Under stringent conditions, no significant DNA homology was detected among the three serotype viruses, except in a restricted portion of these viral genomes. Northern blot hybridization experiments suggested that virus-specific polyadenylated RNA of about 2.4 kilobases was transcribed from a restricted portion showing close homology in these viruses. Southern blot hybridization under less stringent conditions revealed that regions with weak homology were distributed over most of the viral genomes of the three serotypes. Two types of virus-specific glycoproteins, gA and gB, were identified in the immunoprecipitates of the culture medium and cell lysates, respectively, of serotype 2-infected cultures with monoclonal antibodies or hyperimmune antisera cross reactive with serotype 1 MDV and serotype 3 HVT, and detected on the surface of serotype 2-infected cells by the membrane immunofluorescence test. These results indicate a close evolutionary relationship among these three viral serotypes.

Animals↗
PubMed1986

Antigenic differences among multiply charged Moloney murine leukemia virus p30 polypeptides found inside infected cells.

At least three Moloney murine leukemia virus (M-MuLV) p30 polypeptides (p30's), viz., a major species at pI 6.3 and two minor ones at pI 6.1 and pI 6.6, have previously been identified in purified virions by 2-dimensional gel electrophoresis and chromatofocusing (Katoh, I., Yoshinaka, Y. and Luftig, R.B. (1984) J. Gen. Virol. 65, 733-741). We have observed a similar, but distinctive pI pattern for [35S]methionine-labeled MuLV p30's in lysates from chronically infected (MuLV) cells. The variation in pI pattern of the intracellular MuLV p30's was dependent on the type of p30 reactive antibody used for immunoprecipitation. Specifically: a p30 spot with pI 6.3 was always precipitated as the major spot with three different antibodies, minor spots with pI 6.0 and 6.6 were variably seen dependent on the antibody used, and an intracellular p30 spot at pI 6.1 was only precipitated with a rat p30 monoclonal antibody but not with monospecific mouse or intact MuLV cross-reacting p30 sera. These results indicate that first, there are differences between the pI pattern of virion and intracellular MuLV p30's, and second, the antigenic determinants of intracellular p30's vary dependent on the antibody used for immunoprecipitation.

Animals↗
PubMed1986

Experimental study on secondary amyloidosis associated with chronic arthritis.

Various organs from rabbits immunized with heat-killed E. coli 0:14 to induce chronic polyarthritis resembling rheumatoid arthritis (RA) were examined for amyloid deposition since amyloidosis is a frequent feature of long-standing RA. Amyloids were confirmed mainly by polarizing as well as nonpolarizing light microscopy after Congo red staining and partly by electron microscopy and immunoperoxidase technique. Amyloid deposits after an additional 2 months from the first appearance of arthritis were most frequently found in the spleen and kidneys suggesting secondary amyloidosis. Amyloidosis was observed in 36% of 75 arthritic rabbits. The incidence of amyloidosis was significantly higher in the arthritic rabbits than in the non-arthritic rabbits (p less than 0.05). This suggests that prolonged sensitization with heat-killed E. coli 0:14 containing endotoxic substance participates in the formation of both arthritis and amyloidosis.

Amyloidosis↗
PubMed1986

Identification with monoclonal antibodies of virus-specific DNA-binding proteins in the nuclei of cells infected with three serotypes of Marek's disease virus-related viruses.

Two groups of virus-specific polypeptides were identified in the nuclei of infected cells by cross-reacting monoclonal antibodies with three serotypes of Marek's disease virus. Of these, a 135,000-molecular-weight polypeptide common to all three serotypes was found to bind to both double-stranded and single-stranded DNAs.

Animals↗
PubMed1986

Copy number of tandem direct repeats within the inverted repeats of Marek's disease virus DNA.

We previously reported that DNA of the oncogenic strain BC-1 of Marek's disease virus serotype 1 (MDV1) contains three units of tandem direct repeats with 132 base pair (bp) repeats within the inverted repeats of the long regions of the MDV1 genome, whereas the attenuated, nononcogenic viral DNA contains multiple units of tandem direct repeats (Maotani et al., 1986). In the present study, the difference in the copy numbers of 132 bp repeats of oncogenic and nononcogenic MDV1 DNAs in other strains of MDV1 was investigated by Southern blot hybridization. The main copy numbers in different oncogenic MDV1 strains differed: those of BC-1, JM and highly oncogenic Md5 were 3, 5 to 12 and 2, respectively. The viral DNA population with two units of repeats was small, but detectable, in cells infected with either the oncogenic BC-1 or JM strain. The MDV1 DNA in various MD cell lines contained either two units or both two and three units of repeats. The significance of the copy number of repeats in oncogenicity of MDV1 is discussed.

Animals↗
PubMed1985

Origin of human T-lymphotrophic virus I-positive T cell lines in adult T cell leukemia. Analysis of T cell receptor gene rearrangement.

Using the clone-specific rearrangement of the T cell receptor gene as the genetic marker of the clonotype, we analyzed the clonal origin of the interleukin 2 (IL-2)-dependent human T-lymphotrophic virus I (HTLV-I)-positive T cell lines established from various adult T cell leukemia (ATL) patients. From a patient with chronic ATL, whose leukemic cells proliferated in vitro in response to IL-2, we repeatedly established leukemic T cell clones having the same rearrangement profile of the T beta chain gene as the leukemic cells. By contrast, established cell lines from acute ATL patients had different beta chain gene rearrangements from those of the leukemic cells. These HTLV-I+ T cell lines might not be the direct progeny of the leukemic cells, but that of T cells infected either in vivo or in vitro. These IL-2-reactive nonleukemic T cells might have been selected in vitro, because their leukemic cells failed to respond to IL-2, despite the expression of IL-2 receptor. The analysis of the T cell receptor gene rearrangement may give a new approach for the elucidation of the mechanism of leukemogenesis and the origin of the HTLV-I+ T cell lines in ATL.

Acute Disease↗
PubMed1985

Rat lymphoid cell lines producing human T cell leukemia virus. II. Constitutive expression of rat interleukin 2 receptor.

Three rat lymphoid cell lines (TARS-1, TARL-2, and TART-1) (12) transformed by human T cell leukemia/lymphoma virus I (HTLV-I) had rearrangement of the beta chain gene of the T cell antigen receptor, and had integrated proviral DNA from HTLV-I in their genomes. As is the case with adult T cell leukemia (ATL)-derived human T cell lines transformed by HTLV-I, these rat cell lines unequivocally expressed interleukin 2 (IL-2) receptor, as determined by radiolabeled IL-2 binding. By Scatchard plot analysis, one of the cell lines, TART-1, proved to have high affinity receptors (Ka = 1.3 X 10(11)/M and 8.8 X 10(9)/M). Rat IL-2 receptor, not human IL-2 receptor, was expressed on HTLV+ rat cell lines, as demonstrated by the fact that they expressed antigens reactive with monoclonal antibodies (ART-18) against rat IL-2 receptor, but not with anti-Tac antibodies. The collective evidence indicates that the endogenous IL-2 receptor gene is activated in human and rat lymphoid cell lines with HTLV-I production. The mechanism of abnormal IL-2 receptor expression in HTLV infection is discussed.

Animals↗
PubMed1985

Identification of Marek's disease virus-specific antigens in Marek's disease lymphoblastoid cell lines using monoclonal antibody against virus-specific phosphorylated polypeptides.

For identification of the antigens specific to Marek's disease virus (MDV) in virus-non-producer lymphoblastoid cell lines established from a tumor of Marek's disease (MD), hybridomas producing monoclonal antibodies (MAbs) against the antigens were isolated. Immunogens for preparation of the hybridomas were purified from the lysate of an MD-lymphoblastoid cell line, MSB1, by affinity chromatography coupled with chicken anti-MDV serum immunoglobulin G. Three of the MAbs obtained, MB1, MB2 and MB3, were specific to MDV by immunofluorescence test. An immunofluorescence test using MB2 antibody showed that immunofluorescence-positive cells in non-producer MD-lymphoblastoid line cells became detectable when the culture temperature was shifted from 41 degrees C to 33 degrees C or when treatment with 5-iodo-2-deoxyuridine (IUdR) was performed, indicating that the antigen reactive with MB2 antibody is an MDV-specific early antigen. This temperature shift or IUdR-treatment did not induce other MDV-specific antigens, such as late gene products of MDV, detected with MAbs. MB1 and MB2 antibodies immunoprecipitated 4 MDV-specific phosphorylated polypeptides with molecular weights (MWs) of 43,000 (43kd), 39kd, 36kd and 24kd from chick embryo fibroblasts productively infected with virulent MDV. In the place of 43kd, phosphorylated 44kd polypeptide was precipitated from avirulent MDV-infected fibroblasts. However, MB3 antibody did not precipitate any MDV-specific polypeptides from infected fibroblasts. These results suggest that the phosphorylated polypeptides are MDV-specific polypeptides predominantly expressed in non-producer MD-lymphoblastoid cell lines.

Animals↗
PubMed1985

Induction of chronic polyarthritis in rabbits by hyperimmunization with Escherichia coli. I. Pathologic and serologic features in two breeds of rabbits.

Hyperimmunization of 147 rabbits (outbred Japanese white rabbits and New Zealand white [NZW] rabbits bred in a closed colony) with heat-killed Escherichia coli 0:14 in Freund's incomplete adjuvant resulted in the animals developing a chronic polyarthritis resembling rheumatoid arthritis (RA). While both Japanese white and NZW rabbits showed a high incidence of the induced arthritis, a higher proportion of NZW rabbits developed the disease, suggesting that genetic influence is important in the development of RA-like illness. This experimental model may be useful for the study of RA.

Animals↗
PubMed1985

Low frequency of somatic mutation in beta-chain variable region genes of human T-cell receptors.

We have cloned three pairs of rearranged and germ-line variable region (V beta) genes of the beta chain of the human T-cell receptor from the cell lines ATL2, ATL12, and MT-1 of patients with adult T-cell leukemia (ATL). The pairs were derived from the same (for ATL2 and ATL12) and different (for MT-1) individuals. Comparison of the nucleotide sequences showed no somatic mutation in V beta X ATL2 and beta X ATL12-2. Although one nucleotide change was found in V beta X MT1-1, the possibility of polymorphism was not excluded. These results indicate that the frequency of somatic mutation in the V beta gene of the T-cell receptor is 1/10th or less than that in the immunoglobulin gene. Both alleles of the rearranged T-cell receptor gene were analyzed for ATL12 and MT-1. In both, only one of the two rearranged J beta alleles was an active variable-diversity-joining (V-D-J) complex. The results suggest that allelic exclusion occurs in the T-cell receptor gene.

Alleles↗
PubMed1985

Differences in the processing of secreted glycoprotein A induced by Marek's disease virus and herpesvirus of turkeys.

The steps from the precursor to the processed forms of a secreted glycoprotein (gA) of Marek's disease virus (MDV) and of herpesvirus of turkeys (HVT) were examined by two-dimensional gel electrophoresis of immunoprecipitates from the cell lysate and medium of infected cultures with monoclonal antibodies. Differences between MDV- and HVT-gA were particularly observed in isoelectric points of the glycosylated or unglycosylated precursor forms.

Animals↗