Delayed hypersensitivity cross-reactions among Sporothrix species in sporotrichotic patients.

Cutaneous delayed hypersensitivity to antigens prepared from Sporothrix schenckii, S. schenckii var. luriei (atypical S. schenckii), S. curziconia, S. inflata and S. ghanensis was tested in eleven cutaneous sporotrichotic patients. Cross-reactions were observed in various degrees. Five normal controls were not reactive.

Carbohydrate composition of the phenol-soluble lipopolysaccharides of Citrobacter freundii.

Phenol-soluble lipopolysaccharides were obtained from the interphase and phenol phase fractions of 44% aqueous phenol-extracted Citrobacter species. Upon detailed investigation of C. freundii 8090, the two lipopolysaccharide fractions were found to contain different amounts of lipid A, although qualitative composition was similar. Both contained lipid A, 2-keto-deoxyoctonic acid, heptose, phosphate, d-glucose, galactose, rhamnose, 2-acetamido-2 deoxy-d-glucose, 3-acetamido-3,6-dideoxy-d-glucose, O-acetyl, and trace amino acids. Partially purified phenol-phase lipopolysaccharide partitioned into the phenol-soluble phase when refractionated with 44% aqueous phenol, and was further found to be soluble in 88% phenol, 95% ethyl alcohol, and chloroform-methanol (2:1).

Polysaccharides of type 6 Klebsiella.

Water-extractable type 6 Klebsiella antigens were separated into a type 6-specific acidic polysaccharide and a neutral polysaccharide. The neutral polymer was devoid of type 6 activity although it was serologically active. The type 6-specific polymer contained fucose, glucose, and mannose, and pyruvic, galacturonic, and possibly glucuronic acids. The neutral polymer contained glucose, galactose, and mannose.

Occurrence of glucomuramic acid in gram-positive bacteria.

Analyses of 48 gram-positive bacteria indicate only glucomuramic acid and no galactomuramic acid in cell walls.

Serological cross-reactivity between Sporothrix schenckii and various unrelated fungi.

The serological cross-reactivity of Sporothrix schenckii with various unrelated fungi was investigated by use of immunodiffusion tests. A rabbit anti S. schenckii serum was obtained, which reacted with Cladosporium werneckii, C. carrionii, C. bantianum, Coccidioides immitis, Phialophora jeanselmei, P. gougerotii, P. dermatitidis, Fonsecaea pedrosoi, Aspergillus fumigatus, Histoplasma capsulatum and Trichophyton mentagrophytes, but not with Saccharomyces cerevisiae antigens. The serological determinants responsible for the cross-reactions were suggested to be D-galactosyl residues.

Enhancement of macrophage-mediated tumor cell killing by bacterial outer membrane proteins (porins).

Various microbial products are known to influence the function of mouse peritoneal macrophages. Lipopolysaccharide (LPS) and certain lipid A-associated proteins are known to enhance the tumoricidal effects of macrophages. The purpose of this study was to determine whether porins (outer membrane proteins) of Salmonella typhimurium G30/C21 would influence the activity of macrophages from lipid A-responsive and -unresponsive mice. Porins, extracted by a combined sodium dodecyl sulfate-EDTA method from cell walls, were free of LPS as determined by Limulus amebocyte lysate assay and appeared as a band at approximately 36,000 molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In tumor cell killing assays done under LPS-free conditions, the porins in doses of 1 to 10 ng/ml enhanced the tumoricidal effect of macrophages from bacillus Calmette-Guérin-infected C3H/HeN or C3H/HeJ mice. Protein-free LPS enhanced the tumoricidal activity of macrophages from bacillus Calmette-Guérin-infected C3H/HeN but not C3H/HeJ mice. The tumoricidal-enhancing activity of protein-free LPS was blocked by the lipid A-binding antibiotic polymyxin B sulfate, but the effects of porins were not altered by the polymyxin B sulfate. These results suggest that porins, proteins known to alter membrane function, may alter macrophage function by interaction with macrophage membranes.

Strain variation in composition and molecular size of the capsular polysaccharide of Cryptococcus neoformans serotype A.

The capsule of Cryptococcus neoformans is an important virulence factor. In this investigation capsular polysaccharides (CPSs) were isolated by ethanol precipitation from culture filtrates of C. neoformans serotype A strains 6, 15, 98, 110, and 145. Capsule sizes on India ink examination ranged from barely perceptible (strain 15) to greater than the diameter of the yeast cell (strain 6); the others were intermediate in size. On ion-exchange chromatography on DEAE-cellulose each CPS eluted at 0.2 M NaCl; CPS of strain 15 had two major peaks, designated III and IV. On gel-permeation chromatography CPSs of strains 6, 98, 110, and 145 eluted at the void volume of Sepharose CL-2B in the presence or 0.1 M EDTA, while the CPS of strain 15 eluted in two peaks. Sephacryl S-1000 resolved CPSs of all five strains in the following order, from largest to smallest molecular size: 145 greater than 110 greater than 98 greater than 6 much greater than 15. All five CPSs contained mannose, xylose, and glucuronic acid, while the carboxyl-reduced CPS of strain 110 also contained a large percentage of an inositol-like compound. The CPS of strain 110 contained approximately 30% uronic acid by weight, while the others had 15 to 20%. The composition of peak IV from the CPS of strain 15 resembled those of the other strains; peak III of strain 15 contained a substantial amount of galactose. Each CPS contained less than 0.2% protein by weight. The significant differences in molecular size and sugar composition among CPSs of these strains of C. neoformans serotype A may partially explain strain differences in virulence and biological properties of the organism.

Occurrence of antigenic (species-specific?) partially 3-O-methylated heteromannans in cell wall and soluble cellular (nonwall) components of Coccidioides immitis mycelia.

Skin test-active, phenol-soluble, water-soluble (PSWS) extracts of Coccidioides immitis whole, defatted mycelia were compared with skin test-active, alkali-soluble, water-soluble (ASWS) extracts of mycelial cell walls. Both PSWS and ASWS extracts contained partially 3-O-methylated mannan. Composition analysis of both PSWS and ASWS extracts indicated mannose and glucose as major components, whereas 3-O-methylmannose and galactose were minor constituents. These heteromannans and glucans could be obtained from cell walls by extraction with either mild acid or alkali, but not with aqueous phenol. We therefore conclude that the PSWS extracts which were obtainable from whole mycelia but not from cell walls represent cellular (i.e., periplasmic, membrane associated, or cytoplasmic), nonwall polymers. Both PSWS and ASWS mycelial extracts and their gel filtration fractions reacted with rabbit antispherule serum, but with little cross-reactivity. However, alkali treatment of higher-molecular-weight PSWS extract polymers yielded products that cross-reacted with serological identity with low-molecular-weight components of ASWS extracts of mycelial walls, indicating that cell walls and cellular polymers share antigenic determinants. Finally, gel filtration of PSWS extracts on Sephacryl S-200 and Sepharose CL 4B yielded fractions that reacted with serological identity with one or more components of mycelial cytosol, mycelial culture filtrate and autolysate coccidioidins, and spherulin.

Occurrence of pyruvic acid in capsular polysaccharides from various Klebsiella species.

Capsular polysaccharide materials from several different klebsiella serotypes were demonstrated to contain an alpha-keto acid characterized as pyruvic acid. Linkage to the capsular polysaccharides was shown to be acid labile and alkali stable, suggesting ketosidic rather than ester linkages.

Characterization of 3-O-methylmannose from Coccidioides immitis.

3-O-methylmannose isolated from Coccidioides immitis was characterized by gas chromatographic comparison of derivatives with those of synthetic 3-O-methylmannose. Gas chromatographic behavior of three derivatives of the natural and synthetic sugars was identical.

Fractionation and composition studies of skin test-active components of sensitins from Coccidioides immitis.

Coccidioidin skin-test activities from mycelial culture filtrates and autolysates were partially purified. Major chemical constituents included 3-O-methylmannose, mannose, and amino acids.

Cellular localization of uridine 5'-diphosphate-N-acetyl-D-glucosamine oxidoreductase activity in Citrobacter freundii.

The uridine 5'-diphosphate-N-acetyl-d-glucosamine oxidoreductase activity of Citrobacter freundii ATCC 10053 was found to be located in the soluble cytoplasmic fraction of lysed spheroplasts.

Degradation of 14C-labeled streptococcal cell walls by egg white lysozyme and lysosomal enzymes.

The resistance of native and trypsin-treated [14C] glucose-labeled cell walls to degradation by lysozyme and human lysosomal enzymes was confirmed. In contrast, chemically N-acetylated cell walls undergo significant degradation by these enzymes in the pH range of 4.5 to 5.5 without prior removal of the group-specific carbohydrate. N-acetylation after removal of the group A carbohydrate by formamide extraction renders the cell walls considerably more susceptible to these enzymes than by formamaide extraction alone. It appears, therefore, that unless N-acetylation can occur in vivo, streptococcal cell walls are minimally degraded, if at all, by human peripheral blood leukocytes or lysozyme. Examination of leukocyte extracts from normal subjects and patients with post-streptococcal syndromes revealed no qualitative differences in ability to dissolve streptococcal cell walls.

Serological cross-reactivity between group B Streptococcus and Sporothrix schenckii, Ceratocystis species, and Graphium species.

Serological cross-reactivity of a group B Streptococcus (H36B) with Sporothrix schenckii and 39 different Ceratocystis and Graphium species was investigated by double immunodiffusion. Rabbit anti-H36B serum reacted with antigens from S. schenckii and from 36 of 39 Ceratocystis and Graphium species. It is speculated that low-titer agglutinins to S. schenckii in normal sera are due to antibodies raised against various bacteria which share common antigens with S. schenckii.

Identification of products of the uridinediphospho-N-acetyl-D-glucosamine oxidoreductase system from Citrobacter freundii ATCC 10053.

Uridinediphospho-N-acetyl glucosamine is converted to one or more uridine-diphospho-4-keto-6-deoxy-2-acetamidohexose derivatives by an enzyme from Citrobacter freundii ATCC 10053. Borohydride reduction of reaction products followed by acid hydrolysis yielded several amino sugars. Two of these were identified as fucosamine and quinovosamine by chromatography and ninhydrin degradation.