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Biomedical subjects

M Taniguchi

Publications and source records attributed to M Taniguchi.

At least 793 records · Page 44Linked to original sources

Specific suppressive factors produced by hybridomas derived from the fusion of enriched suppressor T cells and a T lymphoma cell line.

A cell fusion technique was used to produce hybridomas between the T lymphoma cell line, EL-4, derived from C57BL (H-2(b)), and an enriched population of human gamma globulin (HGG)-specific suppressor T cells prepared from the spleens of HGG-tolerant CBA mice (H-2(k)). Membrane fluorescence analysis of the hybridoma cells within 6 wk of cell fusion revealed expression of H-2(k) and I-J(k) gene products as well as H-2(b) antigens. Sonicates prepared from hybridomas which contained I-J(k) cells were tested for suppressive activity in vivo in irradiated mice given HGG-primed cells, dinitrophenyl (DNP)-primed cells, HGG-DNP, and horse erythrocytes. Among 18 such hybridoma lines, 6 showed specific suppressive activity, 5 nonspecific suppression, and 7 no suppression. Most lines progressively lost, with time, those properties derived from the normal parent cell. By about 3 mo after fusion few cells expressed CBA markers and only one cell line (number 77) retained some specific suppressive activity. In parallel with the losses was an alteration in chromosome number from near-tetraploid, soon after cell fusion, to near- diploid. Preliminary results with the T lymphoma-sensitive hypoxanthine aminopterin thymidine cell line, L5178, indicate retention of the expression of surface markers derived from the normal parent for 18 wk after hybidization. This suggests that T lymphoma cell lines may have to be screened for their capacity to produce hybridomas with stable properties.

Antibody Formation↗

Ultraviolet flow dichroism of brain microtubule.

Ultraviolet flow dichroism of brain microtubule was measured. Large positive dichroisms were observed at 255 nm, 285 nm and 292 nm. The dichroism at about 255 nm is due to bound GDP or GTP. The dichroism at 285 nm is due to tyrosine and tryptophan residues, and that at 292 nm is due to the tryptophan residues of the protein. These results show that the electronic transition moments of nucleotides and aromatic groups are both nearly parallel to the polymer axis.

Animals↗

Effects of riboflavin deficiency on the lipids of rat liver mitochondria and microsomes.

Weanling male rats were fed a riboflavin deficient diet for 5 weeks and enlargement of the liver mitochondria and discontinuity of the outer membrane were observed. The content of phospholipids was slightly increased in the deficient mitochondria, and decrease in phosphatidylcholine and increase in phosphatidylethanolamine were shown respectively. In comparison with the mitochondria, the content and distribution of phospholipids in microsomes were not affected. The fatty acid composition of phosphatidylcholine in mitochondria and microsomes was remarkably altered by the deficiency, and increases in palmitic and linoleic acids and decrease in arachidonic acid were demonstrated. The incorporation of 32P into diphosphatidylglycerol in mitochondria was reduced by the deficiency. The incorporation into other phospholipids was not significantly altered, whereas the incorporation into the subspecies of phosphatidylcholine was variously affected. By the intraperitoneal injection of riboflavin to the deficient rats, normalization of the mitochondrial size and fatty acid composition of liver mitochondrial lipids was observed. However, decreased incorporation of 32P into diphosphatidylglycerol in mitochondria was not recovered COMPLETELY AT 40 Hours after the injection, and in the mitochondrial lipids linoleic acid was higher and arachidonic acid was lower than respective controls at 60 hours.

Animals↗

Specific suppression of the immune response by a factor obtained from spleen cells of mice tolerant to human gamma-globulin.

An antigen-specific suppressive factor was extracted from spleen cells of mice made tolerant by injection of deaggregated human gamma-globulin (HGG). The active material could be prepared from T cells, obtained by passaging spleen cells through an anti-immunoglobulin column, although not from cells adherent to the column nor from spleen cells pretreated with anti-Thy-1 serum and C. This factor was antigen-specific since it was retained on immunoadsorbents containing HGG, but not on columns coated with antibody to HGG or with irrelevant antigens. Despite its specificity for antigen it did not bear any classical immunoglobulin determinants. Its m.w. ranged between 30,000 and 55,000 daltons. It was a product of the I region of the major histocompatibility complex since it carried Ia determinants. The properties of this factor are very similar to those reported elsewhere for suppressive factors obtained from primed T cells, cells from nonresponder mice, and allotype-specific cells. This suggest the existence of a major class of immunoregulatory molecules, nonimmunoglobulin in nature, and responsible for the mediation of antigen-specific T cell-dependent suppression.

Animals↗

Enrichment of specific suppressor T cells and characterization of their surface markers.

A simple method is described which allows antigen-specific suppressor T cells to be enriched by greater than 100-fold. The enriched cells have the following characteristic markers: Ig-, Thy-1+, Ly-1-, Ly-2,3+, and I-J+. More than 30% of this population could be killed directly by an antiserum specific for the I-J subregion gene product in the presence of complement.

Animals↗

Dynamic characteristics of F-actin and thin filaments in vivo and in vitro.

Measurements of birefringence, ultraviolet dichorism and quasielastic light scattering were carried out on F-actin in solution and on the thin filaments of glycerinated myofibrils. The birefringence of the I-bands of myofibrils was of the same order of magnitude as that of F-actin or the F-actin-tropomyosin-troponin complex oriented in vitro at the same concentration. The ultraviolet dichroism spectrum of the I-bands was very similar to that of F-actin or the F-actin complex in vitro, which is due to orientation of bound ADP and tryptophan residues in F-actin. Quasielastic light scattering measurements, electronmicroscopic observations and the analyses of the electro-optic effect of the I-bands suggested approximately the same flexibility for F-actin in vitro and for the thin filaments in vivo. These optical measurements which were made under various conditions provide evidence for a conformational change induced by calcium ions in F-actin both in vivo and in vitro. This conformational change was found to be amplified by the interaction of F-actin with myosin. This is a brief review of our investigation on the dynamics of F-actin and the thin filament in vivo and in vitro by optical methods.

Actins↗

Properties of the antigen-specific suppressive T-cell factor in the regulation of antibody response of the mouse. IV. Special subregion assignment of the gene(s) that codes for the suppressive T-cell factor in the H-2 histocompatibility complex.

The locus of the gene that codes for the antigen-specific suppressive T-cell factor was determined to be in a new subregion "I-J" which locates between I-B and I-C subregions in the H-2 histocompatibility complex. This was shown by two different lines of evidence: (a) The absorbing capacity for the suppressive T-cell factor of several alloantisera against restricted I subregions did not correlate with their specificity for previously known Ia molecules which are coded for by genes in I-A and I-C subregions, but was associated with the specificity for the products of genes putatively present between I-B and I-C subregions. By the occurrence of special recombinant strains, i.e. B10.A(5R), B10.A(3R), B10.S(9R), and B10.HTT, which differ with respect to the I-J subregion, we were able to produce alloantisera which distinguish I-J subregion gene products. The absorption studies using these special alloantisera directed to I-J subregion clearly indicated that the suppressive T-cell factor is a product of I-J subregion gene(s), and that the molecule is distinct from known Ia molecules expressed on splenic B cells. (b) Taking advantage of the fact that there is a strict histocompatibility requirement for the effective suppression between the donor and recipient strains of the suppressive T-cell factor, we were able to determine the required identities of the genes in the H-2 complex existing among those present between I-B and I-C. Again, utilizing the T-cell factors obtained from special recombinant strains, i.e. B10.A(4R) and B10.A(5R), we were able to locate the gene that codes for the suppressive T-cell factor reactive only with relevant haplotype strains between I-B and I-C subregions. These results are most reasonably explained by the presence of a new subregion I-J which is specialized in coding for the suppressive T-cell factor as a different molecule from previously known Ia molecules.

Animals↗

Properties of the antigen-specific suppressive T-cell factor in the regulation of antibody response of the mouse. III. Dual gene control of the T-cell-mediated suppression of the antibody response.

The antigen-specific suppressive T-cell factor of mice, which had previously been shown to be an I region gene product, could effectively suppress the in vitro secondary antibody response of spleen cells from syngeneic or H-2 compatible mouse strains but not that of H-2 incompatible strains. The identities among genes in the left side half (K, I-A, and I-B) of the H-2 complex between the donor and recipient strains were found to be both necessary and sufficient for the induction of suppression. This suggests that the acceptor site for the suppressive T-cell factor is also determined by the gene present in the left side half of the H-2 complex. The cell type which expresses the acceptor site was found to be a subset of T cell. In general, the suppressive T-cell factor obtained from F1 mice could suppress the responses of both parental strains, and the parental factors could suppress the response of F1 mice. The results indicate that both suppressor and acceptor molecules are codominantly expressed on F1 T cells. There were found two types of defects in the expression of suppressor and acceptor molecules among mouse strains: A/J mice could not produce the suppressive T-cell factor despite that they could accept the factor produced by other H-2 compatible mouse strains. In contrast, all the B10 congeneic lines could produce the T-cell factor, but could not accept the factor produced by syngeneic and H-2 compatible non-B10 congeneic lines. The F1 hybrid of A/J and B10. A could both produce and accept the T-cell factor, and thus the expressions of suppressor and acceptor molecules were found to be dominant traits. These results indicate that the antigen-specific T-cell-mediated suppression is regulated by at least two genes both present in the H-2 complex, and that the complementation of these two genes is required for the induction of suppression.

Animals↗

Diphasic transformations of F-actin. Effects of urea and MgCl2 on F-actin.

Asakura, Taniguchi and Oosawa [1]proposed that muscle actin polymer under sonic vibration is in a different state from that of the ordinary double stranded helical structure (F-actin), characterised by partially interrupted structures of F-actin, a state of "f-actin". In order to confirm different states for actin polymers [1, 2], physicochemical studies were made by measurements of viscosity, flow birefringence, electric birefringence, fluorescence, electron microscopy, quasielastic light scattering and ATP splitting. The following results were obtained. (1) F-actin polymers can undergo two processes of depolymerization upon treatment with urea and various salts as judged by measurements of flow birefringence and viscosity: one is a rapid process in a solution containing K+ or Ca2+ and urea; the other is a slow process following a brief rapid one in a solution containing Mg2+ and urea. (2) In the presence of Mg2+ and a suitable concentration of urea, F-actin (FMU-actin) appeared to exhibit different properties than ordinary F-actin; it had lower viscosity and lower flow birefringence and it had on the whole a more flexible polymer structure, also judging from experiments of quasielastic light scattering, electric birefringence. The different structure was confirmed directly be electron microscopic observation. The aromatic side chains of FMU-actin were also more mobile than those of F-actin judging from fluorescence measurements. The transformation between F-actin and FMU-actin was reversible. (3) The state of FMU-actin polymers was also characterized by ATP splitting; FMU-actin split about one mole of ATP into ADP and inorganic phosphate per mole of actin monomer at room temperature, where F-actin did not. A molar excess of Mg2+ with respect to actin monomer at room temperature, where F-actin did not. A molar excess of Mg2+ with respect to actin monomer is required for ATP splitting. F-actin in solutions containing K+ or Ca2+ and urea did not split ATP. FMU-actin activated on Mg-ATP-ase of myosin at nearly the same rate as that of F-actin. (4) We have postulated a flexible filament model (f-actin). The relationships between the structure of f-actin and its functional role for force generation during contraction are discussed.

Actins↗

Studies on the composition of phospholipids in rat small intestinal smooth muscle.

The phospholipid composition of rat small intestinal smooth muscle was investigated in comparison with those of the mucosa and liver. Phospholipid content per g of the wet smooth muscle was almost identical with that of the mucosa and was about 1/4 of that in the liver. The phospholipid/protein ratio of the smooth muscle was about 1/2 of the value in the liver. Sphingomyelin content was significantly high and amounted to 18% of total phospholipids. This value was about twice that in the mucosa and 4 times higher than that in the liver. On the other hand, the percent distribution of phosphatidylcholine was lowest in the smooth muscle. Distribution patterns of phosphatidylserine and phosphatidylinositol in the smooth muscle as well as in the mucosa were different from those in the liver. The occurrence of vinyl-ether and ether phospholipids was clearly demonstrated in the smooth muscle as well as in the mucosa. A major part of the ether lipids was detected in the phosphatidylethanolamine fraction, in which they amounted to about 50%; 40% as alkenyl-acyl type and 12% as alkyl-acyl type. A high content of ether lipids was also observed in the phosphatidylethanolamine fraction from mucosa, but the distribution was reversed, that is, 14% alkenyl-acyl type and 28% alkyl-acyl type. Fatty aldehydes, fatty alcohols, and fatty acids were also determined by gas-liquid chromatography. The compositions of fatty aldehydes in the phosphatidylethanolamine fraction from smooth muscle and from mucosa were similar, whereas the compositions of long chain fatty alcohol and fatty acids were clearly different. The compositions of fatty alcohols and fatty acids of the phosphatidylcholine fraction from smooth muscle showed significantly different patterns from those of the phosphatidylethanolamine fraction and from those of the same phospholipid fraction in the mucosa.

Aldehydes↗

The role of receptors for T cell products in antibody formation.

Immunocompetent cell interactions are achieved via direct contact between functionally different cell types or via interactions between soluble factors elaborated by regulatory T cells and specific receptors on responding cells for the T cell factors. In either case, there exist certain restrictions with respect to the effective interactions, which depend on the state of differentiation and genetic background of the responding cell type. Such restrictions are considered to be mainly determined by the development and nature of the receptor site on responding cell types for different T cell factors, which is now refered to the "acceptor" for the T cell factors. The presence of such acceptor sites on different populations of both T and B cells has been demonstrated in various experimental systems, and they are now considered to be the site by which responding cells receive appropriate signal for destination of their further differentiation. We have tried to review the nature and possible role of acceptor sites on both B and T cells for different T cell factors with respect to the induction and regulation of immune responses. A special emphasis was put on the genetic nature of the acceptor site. The observed genetic restrictions in the acceptance of T cell factors by responding cells suggest that such restrictions are needed for meaningful and unmistakable communications between funcionally different immunocometent cells. Furthermore, the presence or absence of acceptor sites for certain T cell factors is supposed to be a very important factor for determination of the immune responsiveness of animals against certain antigens, and thus in some cases the Ir gene effect may predominantly affect the expression of acceptor site. Possible implications of acceptor site in the regulation of antibody response and in the network of immunocompetent cell interactions are discussed.

Animals↗

Effects of riboflavin deficiency on the lipids of rat liver.

Newly weaned male rats were maintained on a riboflavin-free diet for 5 weeks, and a study was made on the effect of the deficiency upon liver lipids. The content of glyceride in the livers varied among the deficient rats. High contents of glyceride were demonstrated in one-third of the deficient rats, whereas the similar level as that of control was shown in the remaining deficient rats. Contents of phospholipids and relative amounts of individual phospholipids were not altered significantly by the deficiency. Riboflavin deficiency exerted effects on fatty acid components of liver lipids. The composition of fatty acids of triglycerides varied in the deficient rats depending on the content of glycerides. However trends of increase in linoleic acid and decrease in palmitic acid towards fatty liver were observed in the deficiency in comparison with the control. On the other hand, the changes in phospholipid fatty acids were similar in all deficient rats, and the increase in linoleic acid and the decrease in arachidonic acid were brought about by the deficiency compared with controls, respectively. In liver homogenates, the incorporation of 14C-palmitate into triglycerides was higher in the deficient rats irrespective of the presence or absence of fluorde but incorporation into phospholipids was slightly lower in the deficient rats than in control animals.

Animals↗