Bibliothek

Notizen: Five major glycoproteins with anti-B agglutinin activity were isolated from seeds of Euonymus Sieboldiana by a procedure based on precipitation with ammonium sulphate, Sepharose 4B gel filtration, CM- and DEAE-Sepharose chromatography and Sephacryl S-200 gel filtration. The purified glycoproteins each gave a single symmetrical peak on Sephacryl S-200 gel filtration with elution volumes corresponding to molecular weights of approximately 15,000 to 130,000, each forming a single precipitin line on gel diffusion plates with anti-E. Sieboldiana antibody. These anti-B glycoproteins were rich in acidic amino acids without cysteine and methionine and contained about 8-36% carbohydrate, of which galactose, arabinose and glucose were the predominant sugars, with small amounts of glucosamine, rhamnose, fucose, xylose, mannose and ribose. The most anti-B active lectin agglutinated human B red blood cells at a concentration of 2 μg/ml and was strongly inhibited by melibiose. The three other lectins with anti-B agglutinin activity, however, were not inhibited by any galactose-containing glycosides.

Notizen: Thirty-six species of higher plants were found to contain substances in their fruit or seed extracts, which were heat stable and inhibitory for anti-Lea agglutinin. Both preparations partially fractionated from Sophora japonica and Euonymus Sieboldiana were also slightly inhibitory for anti-Leb agglutinin.

Notizen: Fourteen species of higher plants were found to contain materials in their fruit or seed extracts which were heat stable, inhibitory for and capable of absorbing anti-H, -A and -B agglutinins and also lectins. The majority of species exhibiting a high grade of such cross-reactivity belonged to two genera, Viburnum of the Caprifoliaceae and Acer of the Aceraceae. Gel diffusion patterns indicated, however, that the A- and B-like activities of the Viburnum fruits were somewhat distinct serologically from blood group A and B substances of human origin.

Notizen: Lea- and Leb-active substances present in normal human urine were isolated by sequential membrane filtration, column chromatography with DEAE-Sephadex (A-25), and gel filtration of Bio-Gel (P-6). Lea-active substance with the highest activity was obtained in the P-1 fraction (0.8 mg/1) from urine of a non-secretor type by the final gel filtration. Leb-active substance with the highest activity was also obtained in the P-1 fraction (0.2 mg/1) from urine of a secretor type. The P-1 of the non-secretor inhibited Lea-anti-Lea haemagglutination minimally at 4 μg/ml, and the P-1 of the secretor inhibited Leb-anti-Leb haemagglutination at 1 μg/ml. Both P-1 fractions possessing Lea or Leb activity were characteristic in having a much higher fucose content than other isolated fractions, and seemed to be complex carbohydrates with molecular weights of the order of 4–5 × 103.

Notizen: Euonymus Sieboldiana seeds possess a lectin for human erythrocytes with anti-B specificity. Lectin fractions with strong agglutinating activity were separated by ammonium sulphate precipitation from blood group H-like substance, which is coexistent in the same seeds. The lectin has properties resembling a complete agglutinin, being non-dialysable, inactivated by heat treatment at 70°C and specific for D-galactoside.

Notizen: Abstract Intrapleural exudates have been produced in rats and guinea-pigs using calcium pyrophosphate, cell mediated immunity, reversed passive Arthus reaction and carrageenan. The intracellular concentration of various pharmacological mediators has been measured at different times during the evolution of these inflammatory responses. These have been correlated with volume of exudate and cellular migration. The significance of these findings has been discussed and the possible application to the search for new anti-inflammatory agents and their use in diagnostic procedures.

Notizen: Abstract Migration inhibitory (MI) activity in exudates, and ‘migration’ and ‘inhibition of migration’ of exudate cells was investigated in the delayed hypersensitivity (DH) reaction induced by intrapleural injection of PPD into complete Freund's adjuvant(CFA)-sensitized guinea-pigs. During the initial reaction (6-hour), two types of antigen-dependent MI activity were detected in serum and cell free exudate. One was a high molecular weight material associated with immunoglobulin, and the other was a low molecular weight material and appeared to be so-called antigen-dependent migration inhibitory factor (MIF). As the reaction progressed (i.e. 12–24-hour), two types of antigen-independent MI activity were revealed in exudate, but not in serum. One of these was a high molecular weight material, and the other was a low molecular weight material and thought to be socalled antigen-independent MIF. Similar experiments were performed on the reversed passive Arthus (RPA) reaction in the pleural cavity of guinea-pigs. A high molecular weight substance having MI activity was detected in 6-hour cell free exudate and was found to be antigen-independent. So-called MIF was not found in this reaction. Migration of unseparated exudate cells from the 18-hour DH reaction was less extensive than that of 6-hour unseparated cells. Addition of antigen caused further inhibition, the effect on 18-hour exudate cells being more pronounced. These results were further examined, using mononuclear cells separated on a Ficoll-Isopaque gradient. The migration area of the mononuclear cells was reduced as the DH reaction progressed. Mononuclear cells from the DH reaction gave a smaller migration area throughout the reaction in comparison with blood mononuclear cells. The migration area of the RPA exudate mononuclear cells was also reduced as the reaction progressed. However 6-hour RPA exudate mononuclear cells gave a larger migration area than blood mononuclear cells. The migration of mononuclear cells from DH exudate was inhibited by addition of antigen. A greater degree of inhibition of migration was induced by addition of antigen to mononuclear cells from 18- and 24-hour exudate cells in comparison with 6- and 12-hour exudates. The migration of mononuclear cells from normal blood (i.e. unsensitized animals) and RPA exudate (6- and 18-hour) was unaffected by addition of antigen. Similar results were obtained for blood of complete Freund's adjuvant sensitized animals from 0- to 18-hour. Adherent (A) and nonadherent (NA) mononuclear cells from 18-hour DH exudate were separated through a glass bead column. The migration of adherent cells from 6-hour exudates was not significantly inhibited whereas that of 18-hour mononuclear cells was markedly inhibited. The effect on migration of each mononuclear cell fraction was detected by mixing with peritoneal exudate cells (used as indicator cells). The effect of the A cells from 18-hour mononuclear cell exudates on peritoneal exudate cells was strong, whereas that of 6-hour exudates was less marked. A similar effect was observed with NA cells, from 6- and 18-hour exudate, on the migration of peritoneal mononuclear cells.