Search PubMed⌕ Search

Biomedical subjects

T Ohtomo

Publications and source records attributed to T Ohtomo.

65 records · Page 4Linked to original sources

Encapsulation by transformation of strains of Staphylococcus aureus determined by the serum-soft agar technique.

Transformation for capsular-types (A and B) between four Staphylococcus aureus strains was attempted using a serum-soft agar technique to distinguish the capsular-type. DNA preparations from the Smith diffuse strain (capsular-type A) transformed strain NS58C (unencapsulated variant of capsular-type B) to type A. The strain NS58C required Ala- and His-, however, the two transformants tested were Ala+ and His+. These properties coincided with those of the donor strain just as the following characters of the transformants, which are quite different from the untransformed recipient cells: i) negative clumping factor reaction, ii) diffuse-type growth in serum-soft agar, and iii) high mouse virulence. The change to capsular-type A was antigenically comfirmed by electronmicroscopy using ferritin conjugated anti-capsular antibody.

Bacteriological Techniques↗

Mechanism of compact-colony formation by strains of Staphylococcus aureus in serum soft agar.

Compact-colony forming active substance (CCFAS), the material responsible for the compact colonies of Staphylococcus aureus observed in serum soft agar, was found to be an alkaline-stable, associated polysaccharide containing galactose, N-acetylglucosamine, ribitol, phosphorus and a small quantity of alanine. This substance, when extracted from strains unable to produce protein A clumping factor, was able to absorb the serum-reacting factor whereas a teichoic acid preparation of one strain could not. The formation of CCFAS was unaffected by the age of the cells, whereas when staphylococci were cultured at alkaline pH, young cells produced more clumping factor than old ones. Both fibrinogen and its degradation products were capable of inducing compact colonies in a strain of S. aureus. The ability of human sera to interact in compact-colony formation was independent of the immunoglobin content. Thus neither protein A, clumping factor, nor teichoic acid participate in the CCFAS reaction.

Acetylglucosamine↗

Relationship of capsular type to biochemical and immunological properties of teichoic acid preparations from unencapsulated strains of Staphylococcus aureus.

We investigated the biochemical and immunological characteristics of teichoic acid preparations (TAP) obtained from four unencapsulated strains of Staphylococcus aureus which nonetheless, according to the serum-soft agar technique, produced capsular type antigen and were representative of the four types A, B, C, and D. In the agar diffusion test, TAP of each strain produced a single precipitin line only against rabbit antisera corresponding to the homologous capsular type; no lines were observed against antisera to the heterologous capsular type. All TAP were ribitol type except one, glycerol, prepared from a capsular type D strain. Major acetylglucosaminyl residues of TAP from strains having capsular type A and C antigens were attached to the polyribitol phosphate by beta-linkage, whereas TAP from a type B antigen strain had an alpha-linkage; type D antigen was attached to the polyglycerol phosphate by the beta-linkage. Chemical analyses and infrared spectrograms of these TAP further confirmed their heterogeneous nature.

Acetylglucosamine↗

Mouse virulent strain of Staphylococcus epidermidis. Relation of antiphagocytic activity to the protection-inducing antigen.

Using 10(9) or 10(7) colony-forming units of a strain of Staphylococcus epidermidis (strain 1142) in saline or 5% mucin, respectively, 90 to 100% of mice died within 24 to 48 hr after intraperitoneal challenge infection. These organisms gradually multiplied in the peritoneal cavity when injected intraperitoneally into mice, while the mouse avirulent strain (strain 1124) rapidly decreased and no organisms were found there 20 hr after injection. This strain was capable of inducing resistance against challenge with homologous strains. The resistance appeared as early as the first week and disappeared the 4th week after the immunization. However, no resistance was induced with strain 1124 against challenge with strain 1142. Also, hyperimmune rabbit serum prepared with strain 1142 passively protected against challenge with homologous strain in mice. The protective antibody was absorbed out with homologous organisms but not with strain 1124. Subsequently, a surface substance was obtained from strains 1142 or 1124 by the method of Morse. The 1142 surface substance was capable of inducing a resistance against challenge with the homologous strain but not with the 1124 surface substance. Also, this substance absorbed the protective antibody in hyperimmune rabbit serum prepared with the homologous strain but not with the 1124 surface substance nor with the Smith surface antigen extracted from the Smith strain of Staphylococcus aureus. Conversely, the protective antibody in rabbit anti-Smith strain serum against challenge with the homologous strain was absorbed with the Smith surface antigen but not with the 1142 surface substance. In the agar diffusion test, the 1142 surface substance and the Smith surface antigen produced single precipitin lines only against homologous antisera. Biochemical analysis of the 1142 surface substance showed that the substance contained neither nucleic acids nor proteins but is composed of hexosamine, glycerol, phosphorus, alanine, glycine and phenylalanine.

Amino Acids↗

Comparison of cell wall teichoic acid fractions isolated from three different encapsulated strains of Staphylococcus epidermidis.

Teichoic acid preparations extracted from the cell wall of three serologically different encapsulated strains of Staphylococcus epidermidis, ATCC-31432 (capsular type I), SE-360 (capsular type II) and SE-10 (capsular type III) were purified by DEAE-cellulose and Sephadex G-50 column chromatography. The preparations showed immunological heterogeneity by an agar diffusion test. The chemical properties of the cell wall teichoic acid preparations of capsular types I and III were regarded as N-acetyl-glucosaminyl glycerol-phosphate polymers containing N-acetylglucosamine and phosphate at molar ratios of 0.22-1.0 and 0.33-1.0, respectively. The preparation of capsular type II was assumed to be an alpha-glucosyl glycerol-phosphate polymer containing glucose and phosphate at a molar ratio of 0.49-1.0, and it reacted strongly with concanavalin A. Moreover, alanine, glycine, serine and lysine were shown, among these preparations, to be a common amino acid composition. These results indicate that cell wall teichoic acids obtained from these strains were biochemically and immunologically different from each other.

Cell Wall↗

A humanized single-chain Fv fragment with high targeting potential against human malignant gliomas.

A humanized ONS-M21 antibody (hM21) against human medulloblastoma and glioma cells was engineered as a single-chain Fv fragment (scFv), and its ability to internalize into tumor cells was evaluated by conjugation with ricin A. The scFv of hM21 (schM21) was easily purified from E.coli by one-step affinity column chromatography. Purified schM21 bound to a medulloblastoma ONS-76 cell with almost equal antigen-binding activity of hM21-Fab fragment. Furthermore, the schM21-ricin A conjugate inhibited the growth of ONS-76 cells, but not that of antigen-negative hepatoma HuH-7 cells, suggesting that the schM21 can be internalized after binding to antigen-positive cells. Thus, schM21 could be expected to act as a novel carrier of diagnostic and therapeutic agents for brain tumors.

Antibodies, Monoclonal↗