Biomedical subjects

K Ikuta

Publications and source records attributed to K Ikuta.

At least 181 records · Page 10Linked to original sources

PubMed1996

Intracellular glutathione as a possible direct blocker of HIV type 1 reverse transcription.

In AIDS patients, chronic inflammation and elevated levels of cytokines seem to be associated with reduced levels of glutathione (GSH). GSH has been proposed to inhibit the activation of NF-kB, which results in the inhibition of HIV-1 replication. Here, we show the evidence that GSH and N-acetylcysteine, but not L-cysteine or dithiothreitol, could inhibit the reverse transcriptase (RT) process of HIV-1. Such inhibition was not observed with the RT of murine leukemia virus.

Acetylcysteine↗

PubMed1996

Identification of novel lymphoid tissues in murine intestinal mucosa where clusters of c-kit+ IL-7R+ Thy1+ lympho-hemopoietic progenitors develop.

We have revealed that about one and a half thousand tiny clusters, filled with one thousand closely packed lymphocytes, can be found throughout the murine small and large intestinal mucosa. They are located in crypt lamina propria (cryptopatches; CP) and can be first detected at 14-17 d after birth. A large fraction of lymphocytes in CP expresses c-kit, IL-7R, Thy1 and a lymphocyte function-associated antigen, LFA-1, whereas most of them remain CD3-, TCR alpha beta-, TCR gamma delta-, sIgM-, and B220-. The population size of IL-2R alpha+, HSA+ and Pgp-1+ subsets is variable (20-50%) and the composition of CD8+, Ly-1+, and CD4+ subsets is smaller but also variable (3-20%). In the small intestine, CP do not contain cells undergoing apoptosis nor cells bearing RAG-1 molecules, but do contain dendritic stromal cells bearing CD11c/CD18 molecules. The frequency of DNA replicating cells in CP is higher than that in Peyer's patches (PP), is lower than that in the thymic cortex and is almost comparable with that in the thymic medulla. The numbers of CP remain the same in aged mice (> 114 wk) but double after estrogen treatment even though the thymi are attenuated sharply in both conditions. Thus, with respect to histogenesis, lymphocyte composition and tissue level of cellular behavior, neither PP, isolated lymphoid follicles, peripheral LNs, nor thymus are identical with CP. Finally, CP are virtually absent in lamina propria of IL-7R-deficient mice that display a profound reduction in thymic and peripheral lymphoid cellularity. By contrast, CP are present in germ-free mice and in athymic (nu/nu), SCID, TCR beta x delta-/-, RAG-2-/-, PP-deficient (aly/aly), stem cell factor (Sl/Sld) and c-kit (W/Wv) mutant mice. Taking all of these results together, CP are the first identification of gut-associated murine lymphoid tissues where the generation of IL-7-dependent lympho-hematopoietic progenitors for T and/or B cell descendants may start to take place at the age of commencement of weaning.

Animals↗

PubMed1996

Interleukin 7 receptor-deficient mice lack gammadelta T cells.

The interleukin 7 receptor (IL-7R) plays a crucial role in early B- and T-cell development. It consists of a unique a chain and a common gamma chain [IL-2 receptor gamma chain (IL-2Rgamma)]. Gene inactivation of IL-7, IL-7R, and IL-2Rgamma resulted in severe impairment of B and T lymphopoiesis in mice. In addition, IL-2Rgamma-deficient mice lack gammadelta T cells in the skin and have the impaired development of natural killer (NK) cells and intraepithelial lymphocytes. To explore the role of IL-7/IL-7R system in gammadelta T- and NK-cell development, we have generated and analyzed IL-7R-deficient mice. gammadelta T cells were absent from skin, gut, liver, and spleen in the deficient mice. In contrast, alphabeta T and B cells were detected in reduced, but certain, numbers, and NK cells developed normally. The gammadelta T-cell development in fetal and adult thymus was also completely blocked. These results clearly demonstrate that the signal from IL-7R is indispensable for gammadelta T-cell development in both thymic and extrathymic pathways. On the contrary, it is suggested that NK-cell development requires cytokine(s) other than IL-7.

Animals↗

PubMed1996

Existence of novel branched side chains containing beta-1,2 and alpha-1,6 linkages corresponding to antigenic factor 9 in the mannan of Candida guilliermondii.

Isolation of beta-linkage-containing side chain oligosaccharides from the mannan of Candida guilliermondii IFO 10279 strain has been conducted by acetolysis under mild conditions. A structural study of these oligosaccharides by one- and two-dimensional NMR and methylation analyses indicated the presence of extended oligosaccharide side chains with two consecutive beta-1,2-linked mannose units at the nonreducing terminal of alpha-linked oligosaccharides. The linkage sequence present in this mannan, Man beta 1-->2Man alpha 1-->3Man alpha-->, has also been found in the mannan of Saccharomyces kluyveri but not in the mannan of Candida species. Furthermore, these oligosaccharides are branched at position 6 of the 3-O-substituted mannose units as follows. (Carbohydrate sequence in text) Structure 1 and (Carbohydrate sequence in text) Structure 2 The H-1 signals of the mannose units substituted by a 3,6-di-O-substituted unit showed a significant upfield shift (delta delta = 0.04-0.08 ppm) due to a steric effect. The inhibition of an enzyme-linked immunosorbent assay between the mannan of C. guilliermondii and factor 9 serum with oligosaccharides obtained from several mannans indicated that only the oligosaccharides with the above structure were active, suggesting that these correspond to the epitope of antigenic factor 9.

Antigens, Fungal↗

PubMed1996

Demonstration of Borna disease virus RNA in peripheral blood mononuclear cells derived from Japanese patients with chronic fatigue syndrome.

CFS, a recently named heterogeneous disorder, is an illness of unknown etiology. The association of CFS with viral infections has been suggested. A common association between CFS and several viruses examined has not been confirmed. Here, we centered on the possible link between CFS and BDV infection. By nested RT-PCR followed by hybridization, BDV RNA was demonstrated as a clear signal in PBMCs in 3 out of 25 CFS patients. The amplified cDNA fragments were cloned and sequenced. A total of 16 clones were studied. Intra-patients divergencies of the p24 were 2-9%, 3-20%, and 3-11% in the deduced amino acids. Inter-patient divergencies among the 16 clones were 3-24%. Antibodies to recombinant BDV p24 protein were detected in 6 CFS patients including one carrying BDV RNA. Overall, these gave the prevalence of 32% (8/25) in Japanese CFS patients, suggesting that Japanese CFS is highly associated with active infection of BDV, or a related agent.

Adult↗

PubMed1996

Pure red cell aplasia after major ABO-incompatible bone marrow transplantation: two case reports of treatment with recombinant human erythropoietin.

A 34-year-old man with acute myelocytic leukemia (AML: MO) and a 32-year-old woman with AML: M2 developed pure red cell aplasia (PRCA) after receiving a major ABO incompatible bone marrow transplant (BMT). The first patient responded to recombinant human erythropoietin (rhEPO) therapy, while the second did not. The second patient also received methylprednisolone (m-PSL) but developed reticulocytosis and hemolysis after the administration of m-PSL. Plasmapheresis was then performed and the patient promptly recovered from hemolysis and PRCA. We conclude that close attention must be paid when treating PRCA following major ABO-incompatible BMT with rhEPO and m-PSL, as there is always the potential for massive hemolysis.

ABO Blood-Group System↗

PubMed1996

High susceptibility of U937-derived subclones to human immunodeficiency virus type 1 infection correlates with accumulation of unintegrated circular viral DNA.

Our previous report showed that U937-derived subclones were differentiated into at least three types (high, middle, and low types), even in the subclones expressing similar levels of surface CD4, in terms of the kinetics of the appearance of viral antigens and virus production after infection with human immunodeficiency virus type 1 (HIV-1). Here we showed the evidence that high susceptibility to HIV-1 infection, which was confirmed by the profound expression of viral messages and antigens, was exclusively associated with a high number of the unintegrated extrachromosomal form of viral DNA, but not with the amounts of adsorbed virus RNA nor those of integrated DNA form. The difference in the amounts of extrachromosomal form of viral DNA was also observed in the culture with 3'-azido-3'-deoxythymidine (AZT), indicating that the susceptibility is essentially unrelated to multiple infection events. Thus, the susceptibility of U937-derived subclones to HIV-1 infection seems to be affected by the occurrence of specific events involved in the accumulation of unintegrated viral DNA after viral adsorption.

DNA, Circular↗

PubMed1996

Borna disease virus RNA in peripheral blood mononuclear cells obtained from healthy dairy cattle.

We examined natural infection of Borna disease virus (BDV) in 74 healthy dairy cattle in Japan. The seroprevalence determined using recombinant BDV p24 protein was 20.3%. The prevalence of BDV RNA in peripheral blood mononuclear cells (PBMCs) determined by polymerase chain reaction for p24 was 10.8%. Cell fraction of the PBMCs revealed the presence of BDV RNA in the T cell-enriched fraction. Thus, BDV infection is widespread even among healthy cattle.

Animals↗

PubMed1996

Varied prevalence of Borna disease virus infection in Arabic, thoroughbred and their cross-bred horses in Iran.

Borna disease virus (BDV) naturally infects horses and sheep and induces progressive poliomeningoencephalomyelitis. Here, BDV recombinant proteins of the first open reading frame (ORF-I; coding for p40 nucleoprotein) and the second ORF-II (coding for p24 polymerase cofactor) were immunoblotted with plasma derived from 72 healthy (28 Arabic, 17 thoroughbred and 27 cross-bred) race horses at Tehran in Iran to detect anti-BDV antibodies. In addition, their peripheral blood mononuclear cells (PBMCs) were also examined for BDV RNA by a nested reverse transcriptase-polymerase chain reaction (RT-PCR) at ORF-II. The prevalence of BDV antibodies and/or RNA was 41.2% in Arabic, 23.5% in thoroughbred, and 33.3% in cross-bred horses, but only 17.9, 5.9, and 11.1% of them, respectively, showed positive signals for both BDV antibodies and RNA. Especially, cross-bred horses showed a higher prevalence for BDV RNA, which was detected only in females. In addition, significantly higher prevalence for BDV RNA was observed in Arabic males and thoroughbred females. The BDV prevalence did not increase with aging of the horse. Sequencing at the region of BDV derived from Iranian horses revealed a slight difference from those of Japanese horse- and European horse-derived BDVs even in the amino acid residues, although those in the three groups of Iranian horses were quite similar. Thus, the varied prevalence of BDV was observed with the horse strain or sex in Iranian horses, although BDV sequences were very similar among all three groups in Iran compared with those derived from other countries.

Amino Acid Sequence↗

PubMed1996

Serial passage of human immunodeficiency virus type 1 generates misalignment deletions in non-essential accessory genes.

Human immunodeficiency virus type 1 (HIV-1) derived from an infectious molecular clone pNL432 was extensively passaged in tissue culture by repeated rounds of acute infection. We previously showed the natural occurrence of a nonsense mutation in the vpr gene during continued passage of this virus. In this report, we show that two forms of large deletions (561 and 518 base pairs containing short direct repeats at the deletion junctions) occur after passage 50 in the region that spans the vif and vpr open reading frames. One model to explain the occurrence of these deletion regions is that such mutations result from misalignment of the growing point at a limited number of nucleotide positions. Infection of CD4+ T-cells with a recombinant HIV-1 construct containing the same vif to vpr deletion showed virtually no cytopathogenic phenotype. Thus, misalignment deletions at non-essential accessory genes of HIV-1 might be induced during replication, which result in the generation of virus with a low cytopathogenic potential.

Base Sequence↗

PubMed1996

Laparoscopic-assisted creation of a vagina.

The applicability of laparoscopy for colpopiesis using pelvic peritoneum was examined in two women with the Mayer-Rokitansky-Kuster-Hauser syndrome. Using a modification of laparoscopic guidance proved advantageous for safe dissection of the vesicorectal space and accurate suturing of the pelvic peritoneum without laparotomy. A sufficient vaginal cavity was achieved in both patients. This modification of the original procedure, featuring introduction of a laparoscope, thus provides effective and safe conditions for creation of a new vagina.

Adult↗

PubMed1996

Defective B-1 cell development and impaired immunity against Angiostrongylus cantonensis in IL-5R alpha-deficient mice.

We generated interleukin-5 receptor alpha chain (IL-5R alpha)-deficient (IL-5R alpha-/-) mice by gene targeting. The IL-5R alpha-/- mice showed decreased numbers of B-1 cells concomitant with low serum concentrations of IgM and IgG3. They showed no IL-5-induced enhancement of B cell responses to T-independent antigens. The number of alpha beta T cell receptor-positive thymocytes tended to decrease in 3-week-old IL-5R alpha-/- mice, returning to normal by 6 weeks of age. The IL-5R alpha-/- mice produced basal levels of eosinophils, while their bone marrow cells failed to form eosinophilic colonies in response to IL-5. Impaired eosinophilopoiesis in IL-5R alpha-/-mice enhanced the survival of Angiostrongylus cantonensis. These results indicate that IL-5-induced eosinophils serve as potent effector cells in the killing of Angiostrongylus cantonensis in mice.

Angiostrongylus cantonensis↗

PubMed1996

Mitogenic effect of HIV-infected human T cell lines on mouse B cells mediated by surface immunoglobulin.

Following HIV-1 infection, a number of disorders are induced in both normal T and B cells by virus products derived from infected CD4+ T cells. In the present study, we found that HIV-infected, but not uninfected, human T cell lines generated vigorous blastogenesis and proliferation of freshly isolated mouse B cells in a short-term culture. Neither human B cells nor rat B cells showed significant responses to the HIV-infected T cell lines in the present condition. The mitogenic effect of HIV-infected human T cell line requires direct cell-cell interaction between mouse B cells and HIV-infected T cell lines. Since either mitomycin c treatment or paraformaldehyde fixation of HIV-infected T cell lines resulted in complete loss of the mitogenic effect, it seems that de novo synthesized viral products are responsible for this effect. Furthermore, anti-mouse immunoglobulin antibody inhibited completely the B cell stimulation by the HIV-infected human T cell lines. Thus, surface immunoglobulin (sIg) on mouse B cells appears to be an essential molecule which transduces activation signals from HIV-infected human T cells into cytoplasm of the B cells.

Animals↗

PubMed1996

Developmental heterogeneity of V gamma 1.1 T cells in the mouse liver.

Modifications at V-(D)-J junctions increase the diversity of T-cell receptors (TCR). It has been shown that the levels of N-nucleotide insertion at the V-(D)-J junction in TCR transcripts are different between fetal and adult stages. To clarify developmental stages and pathways of gamma delta T cells in the liver, we analysed the nucleotide sequence of V gamma 1.1-J gamma 4 junctions of intra-hepatic lymphocytes (IHL), spleen cells and developing thymocytes from normal and athymic nude mice. The level of N-insertion increased in thymocytes during ontogeny. The percentage of V gamma 1.1-J gamma 4 transcripts with N-insertion was 3% at day 16 of gestation, 42% at newborn, and 89% at 7 weeks. Transcripts from normal IHL showed intermediate levels of N-insertion between those of newborn and adult thymocytes. In contrast the percentage of N-insertion in nude IHL was 47%, and this value was comparable to that of newborn thymocytes. Among the transcripts of normal IHL, the sequences common with nude IHL showed a newborn level of N-insertion (38%), and the remaining sequences showed an adult level (89%). These results suggested the possibility that V gamma 1.1-expressing T cells in IHL might be a heterogeneous population consisting of the cells developed extrathymically as well as the cells developed intrathymically. The V gamma 1.1-J gamma 4 junctions from spleen cells showed less variability than those from IHL and adult thymocytes. It suggested that gamma delta T cells bearing specific V gamma 1.1 TCR develop and/or home in the spleen.

Age Factors↗

PubMed1996

Long-lasting immune response induced by recombinant bacillus Calmette-Guérin (BCG) secretion system.

The recombinant bacillus Calmette-Guérin (rBCG) secretion system utilizing an extracellular alpha antigen of Mycobacterium kansasii (alpha-K) was characterized biochemically and immunologically. The human immunodeficiency virus type 1 (HIV-1) p17gag B cell epitope fused to alpha-K was secreted in extremely large amounts. At least three mice out of seven inoculated with rBCG generated high titres of antibody to the epitope. The long-lasting antibody production persisted more than 14 months.

Amino Acid Sequence↗

PubMed1996

Individual finger forces acting on a grasped object during shaking actions.

Individual finger grip forces acting on a hand-held object were examined during shaking tasks with a five-finger precision grip. The subjects (n = 13) shook a force transducer-equipped grip object (mass = 400 g) in vertical, horizontal, and mediolateral directions at an average movement speed of 33 cm/s (moderate) and 66 cm/s (fast). In addition, grip forces were examined while the subjects (n = 10) held the object in front of the body and walked or ran in place. It was found that the grip forces for all the fingers changed temporally and spatially coupling with the acceleration of the object resulting from shaking. The results suggest that grip force control is accomplished in an active and anticipatory fashion. Regardless of the shaking direction and speed, among the four fingers the absolute grip force in the index finger was largest, followed by the middle, ring, and little finger forces. The index finger therefore plays a primary role in grip force control during shaking. The percent force contribution by each finger varied depending on the direction of shaking. Contributions of the ring and little fingers were larger when shaken in the horizontal and mediolateral directions than they were in the vertical direction. The results suggest that different finger co-ordination is required in relation to shaking direction. Changes in shaking speed from moderate to fast changed the grip forces for all the fingers. During walking and running, grip force control similar to that during active vertical shaking was required to hold the object safely in the hand.

Adult↗

PubMed1996

Induction of apoptosis by protease-defective particle preparations of human immunodeficiency virus type 1 is specific to a subset of U937-derived subclones.

Several recent reports support the hypothesis that apoptosis occurring in leukocytes of human immunodeficiency virus type 1 (HIV-1)-infected individuals is important in progression to AIDS. Specifically, apoptosis of uninfected bystander cells appears critical in the pathogenesis of disease. Here, we present evidence that protease-defective, gp120-containing HIV-1 (L-2) particle preparations specifically induce apoptosis in cells obtained from a subset of promonocytic U937-derived subclones. The rate of apoptosis induction was inversely correlated with the susceptibility of the U937 subclones to wild-type HIV-1 infection. Three types of apoptosis experiments were performed: DNA content analysis by flow cytometry, apoptotic nuclear degradation by fluorescent microscopy and DNA fragmentation analysis by agarose gel electrophoresis. Kinetic analysis revealed that there was a slower induction of apoptosis by L-2 particle preparations than with tumor necrosis factor (TNF)-alpha or anti-Fas antibody. However, there were no significant differences in the initial binding rates of L-2 particles as well as the binding of TNF-alpha or anti-Fas antibody to the U937 subclones. The basal level of protein kinase C activity was higher in high-type subclones compared with low-type subclones. These results suggest that U937 cells can be divided into at least two subpopulations, one that permits a productive HIV-1 infection but is not subjected to L-2 particle preparation-induced apoptosis, while the other poorly replicates HIV-1 and is subjected to L-2 mediated apoptosis, although at a slower rate than found with TNF-alpha or anti-Fas antibody.

Antibodies, Monoclonal↗

PubMed1996

Clonal prevalence of T cells infiltrating into the pancreas of prediabetic non-obese diabetic mice.

The non-obese diabetic (NOD) mouse spontaneously develops T-cell-mediated autoimmune insulitis. We analyzed the clonotypes of T cell infiltrates of the NOD mouse islets using a new method we have developed recently, which consists of RT-PCR amplification of the CDR3 region of the TCR beta chain mRNA and subsequent single-strand conformation polymorphism (SSCP) analysis. NOD mice of 10-32 weeks of age were shown to accumulate oligoclonal T cells in the pancreas. To examine whether each T cell clone stays in a small area of the pancreas or spreads over the whole pancreas, a pancreas was divided into two pieces, which were then subsequently analyzed in a pair by the above PCR-SSCP method. When a pair produces common bands with the same mobility in SSCP gel, they are likely to represent the presence of the same T cell clones between these two parts of the pancreas. Aged mice (24-32 weeks old) with severe insulitis obviously produced more common bands for most of the Vbeta subfamilies than younger mice (10 weeks old) with only periinsulitis. DNA sequencing verified that these common bands have the same TCR junctional sequences, suggesting that they were derived from the same T cell clones. These results suggest that clonal prevalence of T cells infiltrating into the pancreas occurs in the late stage of insulitis development and that a limited number of T cell clones finally predominate over the whole pancreas.

Animals↗