Vox Sang. 29: 221-227 (1975)
On the Specificity of Mushroom Pleurotus ostreatus and Pleurotus spodoleucus Extracts TADAHISA KOGURE Division of Blood Transfusion, Gunma University Hospital, Maebashi
Abstract. Extracts of mushrooms Pleurotus ostreatus and Pleurotus spodoleucus contain hemagglutinins with anti-H specificity similar to those found in Cytisus sessilifotius and Laburnum alpinurn.
Introduction Many reports on phytohemagglutinin or lectin, showinganti-H specificity, have been published up to date. RENKONEN [18] for the first time discovered anti-H agglutinin in seed extracts of Cytisus sessilifolius, Laburnum alpinum and Lotus tetragonolobus. CAZALand LALAURIE [2] found this agglutinin in seed extracts of Ulex europaeus L. (Colmbra), Ulex provincialis Lois (Montpellier), Ulex Jussiae Webb. (Montpellier) and Tetragonolobus purpureus Webb. (Montpellier). Besides, anti-H agglutinins were found in the roots of Ononis spinosa [8], the mushroom Xylariapolymorpha [19] and the seed extracts of Momordica charantia, and Cerastium tomentosum [6, 11. EISLER[3, 41 assumed the presence of a common antigen in Shigella dysenteriae and human red cells because he found in goat serum immunized with this bacillus an agglutinin against human red cells of each group. He called it heterophile human red cell antigen. ISEKIand TSUNODA [I21 isolated, from the earth, Bacillus fulminans, which produced an enzyme which specifically decomposed 0 substance (H substance or EISLER-KAGAYA’S heterophile antigen), and ISEKIand MASAKI[9] reported that when 0
Received: December 4, 1974; accepted: December 17, 1974.
222
KOGURE
substance was acted upon by the B. fulminans enzyme, it lost the H activity, at the same time liberating L-fucose. WATKINS and MORGAN1211 found that anti-H(0) in eel serum was inhibited by a-methyl-L-fucopyranoside and L-fucose and later, MORGAN and WATKINS[I71 reported that anti-H(0) agglutinin in seed extract of Lotus tetragonolobus was inhibited by L-fucose, and especially strongly by a-methyl-L-fucopyranoside, but that anti-H(0) agglutinin in L. alpinum and Cytisus sessilifolius was not inhibited by simple sugars. ISEKIet al. [lo] reported that agglutination of human red cells against agglutinin in Shigella dysenteriae-immunized chickin serum was inhibited and MORGAN1221 observed that lectins from by D-galactose, and WATKINS Laburnum alpinum and Cytisus sessilifolius were most strongly inhibited by N,N’-diacetylchitobiose(O-~-N-acetyl-D-glucosaminyl-(I ---t 4)-N-acetyl-~glucosamine). The present paper reports on anti-H agglutinins which were found in extracts from Pleurotus ostreatus and Pleurotus spodoleucus.
Materials and Methods Extracts. Commercial mushrooms Pleurotus ostreatus and Pleurotus spodoleucus were each pulverized in saline solution with the homogenizer, centrifuged, and the supernatant, which lysed human red cells, but which, after heating at 56°C for 30 min, agglutinated them, was subjected to fractionation. The fraction which precipitated at 30-60% ammonium sulfate saturation, showed the activity. Hereafter this is referred to as P. ostreatus extract. Red cells. Human 0,A,, A,, B, AB and O h red cells were used. O h red cells, frozen in glycerol were supplied by the Department of Legal Medicine, School of Medicine, Gunma University [ll]. Preparation of H-decomposing enzyme and its application. B. fulminans [12, 131 was cultured in ordinary agar at 37°C for 24 h, the cells were suspended in saline solution, and the suspension was incubated at 37°C for 24 h with frequent shaking to disrupt them. The solution was centrifuged at a high speed, and the fraction obtained from the supernatant at 30-60% ammonium sulfate saturation was used as a crude preparation of Hdecomposing enzyme. 1 vol of 0.2 M phosphate buffer (pH 6.5) was mixed with 5 vol saline solution, and in 3 vol of this mixture human 0 red cells to 2% were suspended. To this suspension was further added 1 vol of the H-decomposing enzyme solution. The mixture was incubated at 37°C for 4 h with occasional shaking. Then the 0 red cells in the mixture were washed with saline solution four times to be examined by agglutination and agglutination inhibition tests.
On the Specificity of Mushroom Pleurotus ostreatus.. .
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Sugars. In agglutination inhibition tests, sugars used were D-galactose, D-glucose, L-fucose, D-mannose, L-rhamnose, D-galactosamine, D-glucosamine, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine,lactose, melibiose, salicin, 2 '-fucosyllactose
(Fuc -a(1+2) ...-.+ Gal !.?; Gluc) and lacto-N-tetraose
Since P. spodoleucus extract showed the same specificity as P. ostreatus extract, only the properties of the latter are described below.
Results As shown in table I, P. ostreatus extract agglutinated 0 red cells most strongly, followed by A,, B, A, and A,B red cells in descending order. It did not agglutinate Oh (Bombay) red cells. P. ostreatus extract was inhibited by secretor-type 0 saliva and S. dysenteriae antigen polysaccharide, but not by nonsecretor-type 0 saliva or Diplococcus pneumoniae type XIV polysaccharide. Agglutination inhibition test with sugars revealed that P. ostreatus extract was inhibited most strongly by 2'-fucosyllactose, less strongly by D-galactose and lactose, still less by melibiose, D-galactosamine, N-acetylD-galactosamine, L-rhamnose and lacto-N-tetraose, but not by L-fucose (Table 11). It was found out that H-decomposing enzyme (a-L-fucosidase) from B. fubninans, removed from 0 red cells and secretor-type 0 saliva not only the H activity against anti-H eel serum but also the activity against P. ostreatus extract.
Discussion P . ostreatus extract agglutinated 0 red cells most strongly, A,, B and A, red cells less strongly, and A,B red cells slightly. Since it was inhibited by secretor-type 0 saliva but not by nonsecretor type 0 saliva, it is considered to be anti-H agglutinin. FLORY [5] found that the fraction precipitated from extracts of the seeds of U. europaeus by 30 and 50% ethanol was readily inhibited by glucosides, such as cellobiose and salicin, while the fraction precipitated by 70% ethanol
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224
Table I. Agglutination titers of human red cells by P. ostreatus extract Blood group
Agglutination titers
0 Le(a+b-) 0 Le(a-b+) A1
Aa B AB
Oh
before papain treatment
after papain treatment
1:32 1:32 1: 8 1:16 1:16 1: 2 0
1:256 1:256 1: 64 1 :128 1 :128 1: 16 0
Treatment of erythrocytes with papain was performed using the method of GOLDSMITH [7].
Table IZ. Inhibition of P. ostreatus extract with sugars
Sugars
Agglutination inhibition titer'
D-Galactose D-Glucose L-Fucose D-Mannose L-Rhamnose D-Gdactosamine D-Glucosamine N-acety 1-D-galactosamine N-acety 1-D-glucosamine Lactose Melibiose Salicin 2 '-Fucosyllactose Lacto-N-tetraose
1: 128 0 0 0 1: 32 1: 64 0 1: 64 0 1: 128 1: 64 0 1:2,048 1: 32
Untreated 0 red cells were used as indicator cells with three hemagglutinating doses of P.ostreatus extract. Agglutination inhibition titer indicates maximum dilution of 10% sugar solution which showed complete inhibition of P.ostreatus extract.
On the Specificity of Mushroom Pleurotus osfreafus...
225
was inhibited by L-fucose. MATSUMOTO and OSAWA[I41 elucidated that two fractions were obtained from seed extract of U. europaeus: one is a group of eel serum-type hemagglutinins, which was obtained at 40% ammonium saturation, and which was inhibited by L-fucose, and the other is a group of Cytisus-type hemagglutinins, which was obtained at 40-70% ammonium sulfate saturation, and which was inhibited by N,N’-diacetylchitobiose. YAMAMOTO [23] isolated, from human O(H) substance, H-active oligosaccharide, containing fucose and observed that when it was treated with a-(I-+2)-fucosidase from B. fulminans to liberate fucose, it lost not only the activity to inhibit eel anti-H antibody but also the activity to inhibit antihuman red cell antibody in s. dysenteriae-immunized chickin serum. MATSUMOTO and OSAWA[15] reported that the treatment of human 0-erythrocytes with H-decomposing enzyme (a-L-fucosidase) from B. fulminans destroyed the agglutinability of the cells not only by eel serum-type hemagglutinins but also by Cytisus-type hemagglutinins of U.europaeus. VOAKet al. [20] reported that lectins from C.sessilifolius and L. alpinum contained anti-HI-H agglutinin, the action of which was strongly inhibited by lactose. The present agglutination inhibition test revealed that Pleurotus extract was most strongly inhibited by 2‘-fucosyllactose, and less strongly by lactose and D-galactose. And when 0 red cells were treated with H-decomposing enzyme (a-L-fucosidase) from B. fufminuns, they lost agglutinability not only against anti-H eel serum but also against Pleurotus extract. MATSUMOTO and OSAWA[161 reported that lactose, lacto-N-tetraose and lacto-N-neotetraose were not inhibitory against the eel serum-type hemagglutinins, eel serum and Ufex hemagglutinin I, whereas these oligosaccharides gave weak but definite inhibition against the Cytisus-type hemagglutinins, C. sessififofius, L. alpinum and UIex hemagglutinin I1 and that 2’-fucosyllactose and lacto-N-fucopentaose I were good inhibitors against both types of hemagglutinins. According to them, the comparison of the inhibiting activities of lactose and 2’-fucosyllactose revealed that the presence of fucosyl residue was the cause of the enhancement of the inhibiting activity against Cytisus-type hemagglutinin, and this suggests that a-L-fucosyl residue at C-2 of D-galactose gives rise to a conformational change of the D-galactose residue so as to make the /I-D-galactosyl linkage more accessible to the hemagglutinin. Consequently extracts of mushrooms P. ostreatus and P. spodoleucus contain haemagglutinins with anti-H specificity similar to those found in C. sessilifolius and L. alpinum.
226
KOGURE
Acknowledgements I wish to express my deep gratitude to Prof. FURUKAWA, Dr. KOCHIBE and Dr. KISHI for their kindness to supply me with strain of B. fulminans, bacterial somatic antigen polysaccharide, 2’-fucosyllactose and lacto-N-tetraose. A report of this work was read at the 21st Congress of the Japan Society of Blood Transfusion, Sapporo 1973.
References 1 BIRD,G. W. G. and WINGHAM, J. : Anti-H from Cerastium tomentosum seeds. A comparison with other seeds anti-H agglutinins. Vox Sang. 19: 132-139 (1970). 2 CAZAL,P. et LALAURIE, M. : Recherches sur quelques phyto-agglutinines spkcifiques des groupes sanguins ABO. Acta haemat. 8:73-82 (1952). 3 EISLER,M.: Uber ein gemeinsames Antigen in den Zellen des Menschen und in Shigabazillen. Z. Immunforsch. 67:38-48 (1930). 4 EISLER,M. : Weitere Untersuchungen iiber das Menschenblutantigen in Shigabakterien und sein Verhaltnis zu deren Forssman Antigen. Z. Immunforsch. 70 :48-57 (1931). 5 FLORY,L. L.: Differences in the H antigen on human buccal cells from secretor and non-secretor individuals. Vox Sang. I 1 :137-156 (1966). 6 FUIINUMA, F. : Studies on group-specific phytohemagglutinin from seed of Momordica charanria. (In Japanese.) Acta crim. jap. 27:20-32 (1961). 7 GOLDSMITH, K. : Papain-treated red cells in the detection of incomplete antibodies. Lancet i:76-77 (1955). 8 HERZOG, P. :SpezifischePhytagglutinine aus Ononis spinosa-Wurzeln. Z. Immunforsch. 117: 53-59 (1959). 9 Iseki, S. and MASAKI,S.: Chemical actions of 0- and A-specific enzymes on the respective blood group substances. Gunma J. med. Sci. 4~105-116(1955). 10 ISEKI, S.; SAEKI,N., and FURUKAWA, K. : Immunochemical studies on bacterial blood
group substances. I. Absorption of the blood group specific antibodies in bacterial antisera by simple sugars. Jap. J. Microbiol. 3 :455-459 (1959). H., and TAKIZAWA, H. : Immunological properties of ‘Bombay’ 11 ISEKI,S. ; TAKIZAWA, phenotype. Proc. Jap. Acad. 46: 803-807 (1970). 12 ISEKI, S. and TSUNODA, S.: On a bacterial enzyme which specifically decomposes 0 substance. Proc. Jap. Acad. 28:370-373 (1952). 13 ISEKI,S.; YAMAMOTO, S., and FURUKAWA, K.: A new O(H) substance-decomposing enzyme produced by an aerobic bacterium. 11. Proc. Jap. Acad. 38:57-62 (1962). 1. and OSAWA,T.: Purification and characterization of an anti-H(0) 14 MATSUMOTO, phytoagglutinin of UIex europaeus. Biochim. biophys. Acta 194 :184-1 89 (1969). 15 MATSUMOTO, 1. and OSAWA,T.: Purification and characterization of a Cytisus-type anti-H(0) phytohemagglutinin from Ulex europaeus seeds. Archs Biochem. Biophys. 140:484-491 (1970). 16 MATSUMOTO,I. and OSAWA.T.: On the specificity of various heterologous anti-H hemagglutinins. Vox Sang. 2 1 :548-557 (1971).
On the Specificity of Mushroom Plewotus ostreutus.. .
227
17 MORGAN,W. T. J. and WATKINS,W. M. : The inhibition of the hemagglutinations in plant seeds by human blood group substances and simple sugars. Br. J. exp. Path. 34:94-103 (1953). K. 0.: Studies on hemagglutinins present in seeds of some representatives 18 RENKONEN, of the family of Leguminosae. AM. Med. exp. Fenn. 26:6&72 (1948). 19 SAINT-PAUL, M. : Les hkmagglutinines vkgktales. Transfusion 4 : 3-37 (1961). 20 VOAK,D. and LODGE,T. W.: The demonstration of anti-HI/HI-H activity in seed anti-H reagents. Vox Sang. 20:3&45 (1971). 21 WATKINS, W. M. and MORGAN,W. T. J.: Neutralization of the anti-H agglutinin in eel serum by simple sugars. Nature, Lond. 169 :825-826 (1952). W. M. and MORGAN,W. T. J.: Further observations on the inhibition of 22 WATKINS, blood-group specific serological reactions by simple sugars of known structure. Vox. Sang. 7:129-150 (1962). H. : Enzyme actions to the oligosaccharides isolated from soluble blood 23 YAMAMOTO, group substances by treatment with triethylamine. (In Japanese.) Kitakanto med. J 19:575-600 (1969).
Dr. TADAHISA KOGURE,Department of Legal Medicine, School of Medicine, Gunma University, Muebushi (Japan)
On the Specificity of Mushroom Pleurotus ostreatus and Pleurotus spodoleucus Extracts TADAHISA KOGURE Division of Blood Transfusion, Gunma University Hospital, Maebashi
Abstract. Extracts of mushrooms Pleurotus ostreatus and Pleurotus spodoleucus contain hemagglutinins with anti-H specificity similar to those found in Cytisus sessilifotius and Laburnum alpinurn.
Introduction Many reports on phytohemagglutinin or lectin, showinganti-H specificity, have been published up to date. RENKONEN [18] for the first time discovered anti-H agglutinin in seed extracts of Cytisus sessilifolius, Laburnum alpinum and Lotus tetragonolobus. CAZALand LALAURIE [2] found this agglutinin in seed extracts of Ulex europaeus L. (Colmbra), Ulex provincialis Lois (Montpellier), Ulex Jussiae Webb. (Montpellier) and Tetragonolobus purpureus Webb. (Montpellier). Besides, anti-H agglutinins were found in the roots of Ononis spinosa [8], the mushroom Xylariapolymorpha [19] and the seed extracts of Momordica charantia, and Cerastium tomentosum [6, 11. EISLER[3, 41 assumed the presence of a common antigen in Shigella dysenteriae and human red cells because he found in goat serum immunized with this bacillus an agglutinin against human red cells of each group. He called it heterophile human red cell antigen. ISEKIand TSUNODA [I21 isolated, from the earth, Bacillus fulminans, which produced an enzyme which specifically decomposed 0 substance (H substance or EISLER-KAGAYA’S heterophile antigen), and ISEKIand MASAKI[9] reported that when 0
Received: December 4, 1974; accepted: December 17, 1974.
222
KOGURE
substance was acted upon by the B. fulminans enzyme, it lost the H activity, at the same time liberating L-fucose. WATKINS and MORGAN1211 found that anti-H(0) in eel serum was inhibited by a-methyl-L-fucopyranoside and L-fucose and later, MORGAN and WATKINS[I71 reported that anti-H(0) agglutinin in seed extract of Lotus tetragonolobus was inhibited by L-fucose, and especially strongly by a-methyl-L-fucopyranoside, but that anti-H(0) agglutinin in L. alpinum and Cytisus sessilifolius was not inhibited by simple sugars. ISEKIet al. [lo] reported that agglutination of human red cells against agglutinin in Shigella dysenteriae-immunized chickin serum was inhibited and MORGAN1221 observed that lectins from by D-galactose, and WATKINS Laburnum alpinum and Cytisus sessilifolius were most strongly inhibited by N,N’-diacetylchitobiose(O-~-N-acetyl-D-glucosaminyl-(I ---t 4)-N-acetyl-~glucosamine). The present paper reports on anti-H agglutinins which were found in extracts from Pleurotus ostreatus and Pleurotus spodoleucus.
Materials and Methods Extracts. Commercial mushrooms Pleurotus ostreatus and Pleurotus spodoleucus were each pulverized in saline solution with the homogenizer, centrifuged, and the supernatant, which lysed human red cells, but which, after heating at 56°C for 30 min, agglutinated them, was subjected to fractionation. The fraction which precipitated at 30-60% ammonium sulfate saturation, showed the activity. Hereafter this is referred to as P. ostreatus extract. Red cells. Human 0,A,, A,, B, AB and O h red cells were used. O h red cells, frozen in glycerol were supplied by the Department of Legal Medicine, School of Medicine, Gunma University [ll]. Preparation of H-decomposing enzyme and its application. B. fulminans [12, 131 was cultured in ordinary agar at 37°C for 24 h, the cells were suspended in saline solution, and the suspension was incubated at 37°C for 24 h with frequent shaking to disrupt them. The solution was centrifuged at a high speed, and the fraction obtained from the supernatant at 30-60% ammonium sulfate saturation was used as a crude preparation of Hdecomposing enzyme. 1 vol of 0.2 M phosphate buffer (pH 6.5) was mixed with 5 vol saline solution, and in 3 vol of this mixture human 0 red cells to 2% were suspended. To this suspension was further added 1 vol of the H-decomposing enzyme solution. The mixture was incubated at 37°C for 4 h with occasional shaking. Then the 0 red cells in the mixture were washed with saline solution four times to be examined by agglutination and agglutination inhibition tests.
On the Specificity of Mushroom Pleurotus ostreatus.. .
223
Sugars. In agglutination inhibition tests, sugars used were D-galactose, D-glucose, L-fucose, D-mannose, L-rhamnose, D-galactosamine, D-glucosamine, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine,lactose, melibiose, salicin, 2 '-fucosyllactose
(Fuc -a(1+2) ...-.+ Gal !.?; Gluc) and lacto-N-tetraose
Since P. spodoleucus extract showed the same specificity as P. ostreatus extract, only the properties of the latter are described below.
Results As shown in table I, P. ostreatus extract agglutinated 0 red cells most strongly, followed by A,, B, A, and A,B red cells in descending order. It did not agglutinate Oh (Bombay) red cells. P. ostreatus extract was inhibited by secretor-type 0 saliva and S. dysenteriae antigen polysaccharide, but not by nonsecretor-type 0 saliva or Diplococcus pneumoniae type XIV polysaccharide. Agglutination inhibition test with sugars revealed that P. ostreatus extract was inhibited most strongly by 2'-fucosyllactose, less strongly by D-galactose and lactose, still less by melibiose, D-galactosamine, N-acetylD-galactosamine, L-rhamnose and lacto-N-tetraose, but not by L-fucose (Table 11). It was found out that H-decomposing enzyme (a-L-fucosidase) from B. fubninans, removed from 0 red cells and secretor-type 0 saliva not only the H activity against anti-H eel serum but also the activity against P. ostreatus extract.
Discussion P . ostreatus extract agglutinated 0 red cells most strongly, A,, B and A, red cells less strongly, and A,B red cells slightly. Since it was inhibited by secretor-type 0 saliva but not by nonsecretor type 0 saliva, it is considered to be anti-H agglutinin. FLORY [5] found that the fraction precipitated from extracts of the seeds of U. europaeus by 30 and 50% ethanol was readily inhibited by glucosides, such as cellobiose and salicin, while the fraction precipitated by 70% ethanol
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224
Table I. Agglutination titers of human red cells by P. ostreatus extract Blood group
Agglutination titers
0 Le(a+b-) 0 Le(a-b+) A1
Aa B AB
Oh
before papain treatment
after papain treatment
1:32 1:32 1: 8 1:16 1:16 1: 2 0
1:256 1:256 1: 64 1 :128 1 :128 1: 16 0
Treatment of erythrocytes with papain was performed using the method of GOLDSMITH [7].
Table IZ. Inhibition of P. ostreatus extract with sugars
Sugars
Agglutination inhibition titer'
D-Galactose D-Glucose L-Fucose D-Mannose L-Rhamnose D-Gdactosamine D-Glucosamine N-acety 1-D-galactosamine N-acety 1-D-glucosamine Lactose Melibiose Salicin 2 '-Fucosyllactose Lacto-N-tetraose
1: 128 0 0 0 1: 32 1: 64 0 1: 64 0 1: 128 1: 64 0 1:2,048 1: 32
Untreated 0 red cells were used as indicator cells with three hemagglutinating doses of P.ostreatus extract. Agglutination inhibition titer indicates maximum dilution of 10% sugar solution which showed complete inhibition of P.ostreatus extract.
On the Specificity of Mushroom Pleurotus osfreafus...
225
was inhibited by L-fucose. MATSUMOTO and OSAWA[I41 elucidated that two fractions were obtained from seed extract of U. europaeus: one is a group of eel serum-type hemagglutinins, which was obtained at 40% ammonium saturation, and which was inhibited by L-fucose, and the other is a group of Cytisus-type hemagglutinins, which was obtained at 40-70% ammonium sulfate saturation, and which was inhibited by N,N’-diacetylchitobiose. YAMAMOTO [23] isolated, from human O(H) substance, H-active oligosaccharide, containing fucose and observed that when it was treated with a-(I-+2)-fucosidase from B. fulminans to liberate fucose, it lost not only the activity to inhibit eel anti-H antibody but also the activity to inhibit antihuman red cell antibody in s. dysenteriae-immunized chickin serum. MATSUMOTO and OSAWA[15] reported that the treatment of human 0-erythrocytes with H-decomposing enzyme (a-L-fucosidase) from B. fulminans destroyed the agglutinability of the cells not only by eel serum-type hemagglutinins but also by Cytisus-type hemagglutinins of U.europaeus. VOAKet al. [20] reported that lectins from C.sessilifolius and L. alpinum contained anti-HI-H agglutinin, the action of which was strongly inhibited by lactose. The present agglutination inhibition test revealed that Pleurotus extract was most strongly inhibited by 2‘-fucosyllactose, and less strongly by lactose and D-galactose. And when 0 red cells were treated with H-decomposing enzyme (a-L-fucosidase) from B. fufminuns, they lost agglutinability not only against anti-H eel serum but also against Pleurotus extract. MATSUMOTO and OSAWA[161 reported that lactose, lacto-N-tetraose and lacto-N-neotetraose were not inhibitory against the eel serum-type hemagglutinins, eel serum and Ufex hemagglutinin I, whereas these oligosaccharides gave weak but definite inhibition against the Cytisus-type hemagglutinins, C. sessififofius, L. alpinum and UIex hemagglutinin I1 and that 2’-fucosyllactose and lacto-N-fucopentaose I were good inhibitors against both types of hemagglutinins. According to them, the comparison of the inhibiting activities of lactose and 2’-fucosyllactose revealed that the presence of fucosyl residue was the cause of the enhancement of the inhibiting activity against Cytisus-type hemagglutinin, and this suggests that a-L-fucosyl residue at C-2 of D-galactose gives rise to a conformational change of the D-galactose residue so as to make the /I-D-galactosyl linkage more accessible to the hemagglutinin. Consequently extracts of mushrooms P. ostreatus and P. spodoleucus contain haemagglutinins with anti-H specificity similar to those found in C. sessilifolius and L. alpinum.
226
KOGURE
Acknowledgements I wish to express my deep gratitude to Prof. FURUKAWA, Dr. KOCHIBE and Dr. KISHI for their kindness to supply me with strain of B. fulminans, bacterial somatic antigen polysaccharide, 2’-fucosyllactose and lacto-N-tetraose. A report of this work was read at the 21st Congress of the Japan Society of Blood Transfusion, Sapporo 1973.
References 1 BIRD,G. W. G. and WINGHAM, J. : Anti-H from Cerastium tomentosum seeds. A comparison with other seeds anti-H agglutinins. Vox Sang. 19: 132-139 (1970). 2 CAZAL,P. et LALAURIE, M. : Recherches sur quelques phyto-agglutinines spkcifiques des groupes sanguins ABO. Acta haemat. 8:73-82 (1952). 3 EISLER,M.: Uber ein gemeinsames Antigen in den Zellen des Menschen und in Shigabazillen. Z. Immunforsch. 67:38-48 (1930). 4 EISLER,M. : Weitere Untersuchungen iiber das Menschenblutantigen in Shigabakterien und sein Verhaltnis zu deren Forssman Antigen. Z. Immunforsch. 70 :48-57 (1931). 5 FLORY,L. L.: Differences in the H antigen on human buccal cells from secretor and non-secretor individuals. Vox Sang. I 1 :137-156 (1966). 6 FUIINUMA, F. : Studies on group-specific phytohemagglutinin from seed of Momordica charanria. (In Japanese.) Acta crim. jap. 27:20-32 (1961). 7 GOLDSMITH, K. : Papain-treated red cells in the detection of incomplete antibodies. Lancet i:76-77 (1955). 8 HERZOG, P. :SpezifischePhytagglutinine aus Ononis spinosa-Wurzeln. Z. Immunforsch. 117: 53-59 (1959). 9 Iseki, S. and MASAKI,S.: Chemical actions of 0- and A-specific enzymes on the respective blood group substances. Gunma J. med. Sci. 4~105-116(1955). 10 ISEKI, S.; SAEKI,N., and FURUKAWA, K. : Immunochemical studies on bacterial blood
group substances. I. Absorption of the blood group specific antibodies in bacterial antisera by simple sugars. Jap. J. Microbiol. 3 :455-459 (1959). H., and TAKIZAWA, H. : Immunological properties of ‘Bombay’ 11 ISEKI,S. ; TAKIZAWA, phenotype. Proc. Jap. Acad. 46: 803-807 (1970). 12 ISEKI, S. and TSUNODA, S.: On a bacterial enzyme which specifically decomposes 0 substance. Proc. Jap. Acad. 28:370-373 (1952). 13 ISEKI,S.; YAMAMOTO, S., and FURUKAWA, K.: A new O(H) substance-decomposing enzyme produced by an aerobic bacterium. 11. Proc. Jap. Acad. 38:57-62 (1962). 1. and OSAWA,T.: Purification and characterization of an anti-H(0) 14 MATSUMOTO, phytoagglutinin of UIex europaeus. Biochim. biophys. Acta 194 :184-1 89 (1969). 15 MATSUMOTO, 1. and OSAWA,T.: Purification and characterization of a Cytisus-type anti-H(0) phytohemagglutinin from Ulex europaeus seeds. Archs Biochem. Biophys. 140:484-491 (1970). 16 MATSUMOTO,I. and OSAWA.T.: On the specificity of various heterologous anti-H hemagglutinins. Vox Sang. 2 1 :548-557 (1971).
On the Specificity of Mushroom Plewotus ostreutus.. .
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17 MORGAN,W. T. J. and WATKINS,W. M. : The inhibition of the hemagglutinations in plant seeds by human blood group substances and simple sugars. Br. J. exp. Path. 34:94-103 (1953). K. 0.: Studies on hemagglutinins present in seeds of some representatives 18 RENKONEN, of the family of Leguminosae. AM. Med. exp. Fenn. 26:6&72 (1948). 19 SAINT-PAUL, M. : Les hkmagglutinines vkgktales. Transfusion 4 : 3-37 (1961). 20 VOAK,D. and LODGE,T. W.: The demonstration of anti-HI/HI-H activity in seed anti-H reagents. Vox Sang. 20:3&45 (1971). 21 WATKINS, W. M. and MORGAN,W. T. J.: Neutralization of the anti-H agglutinin in eel serum by simple sugars. Nature, Lond. 169 :825-826 (1952). W. M. and MORGAN,W. T. J.: Further observations on the inhibition of 22 WATKINS, blood-group specific serological reactions by simple sugars of known structure. Vox. Sang. 7:129-150 (1962). H. : Enzyme actions to the oligosaccharides isolated from soluble blood 23 YAMAMOTO, group substances by treatment with triethylamine. (In Japanese.) Kitakanto med. J 19:575-600 (1969).
Dr. TADAHISA KOGURE,Department of Legal Medicine, School of Medicine, Gunma University, Muebushi (Japan)
