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J C Sadoff

Publications and source records attributed to J C Sadoff.

At least 109 records · Page 6Linked to original sources
PubMed1981

Production of exoenzyme S by clinical isolates of Pseudomonas aeruginosa.

Exoenzyme S differs from toxin A and diphtheria toxin in that it does not adenosine diphosphate (ADP)-ribosylate elongation factor-2, but rather catalyzes the transfer of the ADP-ribose moiety of nicotinamide adenine dinucleotide to a number of different proteins in extracts of eucaryotic cells. Polyoma-transformed BHK-21 cells were isolated which were resistant to diphtheria toxin and toxin A. Extracts from these cells are ADP-ribosylated by exoenzyme S but not toxin A or diphtheria toxin, providing an assay which distinguishes between S and A activities. A total of 124 clinical isolates of P. aeruginosa were analyzed for production of toxin A and exoenzyme S. Exoenzyme S production was detected in 38% of the strains, whereas 80% of the strains produced toxin A.

ADP Ribose Transferases↗
PubMed1981

High-molecular-weight polysaccharide antigen from Pseudomonas aeruginosa immunotype 2.

Previously, we isolated a high-molecular-weight immunogenic polysaccharide (designated PS) from Pseudomonas aeruginosa immunotype 1 (IT-1). The method which we used was modified to permit the isolation of a similar PS from P. aeruginosa IT-2. This antigen was composed primarily of carbohydrate, had a complex monosaccharide composition, including sugars not found in the lipopolysaccharide, and was nonpyrogenic in rabbits and nontoxic in mice at high doses. This material protected mice from challenges with live homologous cells. P. aeruginosa IT-2 PS gave a line of identity with the O side chain of the lipopolysaccharide, but different from this polysaccharide in molecular weight, chemical composition, and ability to immunize mice actively. Lipopolysaccharide from P. aeruginosa IT-2 contained an immunological determinant not found on P. aeruginosa IT-2 PS, which was detected due to its stability during treatment with dilute alkali. Thus, we recovered a high-molecular-weight PS antigen from P. aeruginosa IT-2, which was serologically identical to the lipopolysaccharide O side chain but was chemically and physically distinct. Also, like P. aeruginosa IT-1 strains, P. aeruginosa IT-2 contains an alkali-stable immunodeterminant on the lipopolysaccharide that may represent a core-like antigen.

Epitopes↗
PubMed1981

Gonococcal pilus vaccine. Studies of antigenicity and inhibition of attachment.

A gonococcal pilus vaccine or placebo was injected subcutaneously or intramuscularly into 71 human volunteers. The vaccine was found to be safe. The principal adverse reaction was a complaint of a sore arm, which was caused, at least in part, to the volume of material injected. 6 of 64 (9%) volunteers receiving the larger doses also complained of malaise. The vaccine was found to be antigenic. All of the volunteers developed an immunoglobulin class-specific antibody response as measured by a solid phase radioimmunoassay. The antibody was capable of blocking the attachment of gonococci to epithelial cells. A slight antibody response was also demonstrated to gonococcal lipopolysaccharide but the antibody responsible for blocking attachment of gonococci was directed entirely at the pilus protein. The stimulated antibodies were shown to crossreact with isolated pili of heterologous gonococcal strains and to block the attachment of heterologous gonococci. Absorption of immune sera by a heterologous pilus reduced the inhibition of attachment antibodies to pre-immune level, suggesting that the immune response was directed at a common pilus determinant.

Antibodies, Bacterial↗
PubMed1980

Binding of diphtheria toxin to phospholipids in liposomes.

Diphtheria toxin bound to the phosphate portion of some, but not all, phospholipids in liposomes. Liposomes consisting of dimyristoyl phosphatidylcholine and cholesterol did not bind toxin. Addition of 20 mol% (compared to dimyristoyl phosphatidylcholine) of dipalmitoyl phosphatidic acid, dicetyl phosphate, phosphatidylinositol phosphate, cardiolipin, or phosphatidylserine in the liposomes resulted in substantial binding of toxin. Inclusion of phosphatidylinositol in dimyristol phosphatidylcholine/cholesterol liposomes did not result in toxin binding. The calcium salt of dipalmitoyl phosphatidic acid was more effective than the sodium salt, and the highest level of binding occurred with liposomes consisting only of dipalmitoyl phosphatidic acid (calcium salt) and cholesterol. Binding of toxin to liposomes was dependent on pH, and the pattern of pH dependence varied with liposomes having different compositions. Incubation of diphtheria toxin with liposomes containing dicetyl phosphate resulted in maximal binding at pH 3.6, whereas binding to liposomes containing phosphatidylinositol phosphate was maximal above pH 7. Toxin did not bind to liposomes containing 20 mol% of a free fatty acid (palmitic acid) or a sulfated lipid (3-sulfogalactosylceramide). Toxin binding to dicetyl phosphate or phosphatidylinositol phosphate was inhibited by UTP, ATP, phosphocholine, or p-nitrophenyl phosphate, but not by uracil. We conclude that (a) diphtheria toxin binds specifically to the phosphate portion of certain phospholipids, (b) binding to phospholipids in liposomes is dependent on pH, but is not due only to electrostatic interaction, and (c) binding may be strongly influenced by the composition of adjacent phospholipids that do not bind toxin. We propose that a minor membrane phospholipid (such as phosphatidylinositol phosphate or phosphatidic acid), or that some other phosphorylated membrane molecule (such as a phosphoprotein) may be important in the initial binding of diphtheria toxin to cells.

Cardiolipins↗
PubMed1980

Evidence for the role of toxin A in the pathogenesis of infection with Pseudomonas aeruginosa in humans.

Levels of antibody to toxin A of Pseudomonas aeruginosa were determined by a solid-phase radioimmunoassay. Mean (+/- SEM) peak levels of IgG in 24 normal soldiers were 2.6 +/- 0.5 microgram/ml, white mean peak levels in 12 patients colonized with and 13 patients infected in sites other than the blood with toxin A-producing strains were 16.7 +/- 7.0 and 17.1 +/- 4.4 microgram/ml, respectively. Levels of IgG were determined in 52 patients with pseudomonas bacteremia, and those surviving and those dying of bacteremia due to toxin A-producing strains had mean peak levels of 25.8 +/- 5.5 and 4.6 +/- 2.0 microgram/ml, respectively. The antitoxin response in sequential bacteremic sera began shortly after onset of bacteremia and decreased gradually, but antitoxin could be recalled promptly upon reinfection with Pseudomonas. Death from pseudomonas bacteremia was significantly associated with infection by a toxin A-producing strain, presence of underlying disease, hypotension, and antitoxin level of < 2 microgram/ml.

Adult↗
PubMed1980

Toxin A-deficient mutants of Pseudomonas aeruginosa PA103: isolation and characterization.

An immunological assay utilizing double-diffusion principles was developed for identification of Pseudomonas aeruginosa mutants deficient in toxin A. Mutations were chemically induced, and mutants were isolated from P. aeruginosa strain PA103. Quantitative assays, both enzymatic and immunological, indicated that five mutants produced toxin A in vitro at levels of 0.3% or less of parental strain levels. Characterization indicated that the mutants fell into four classes and suggested that multiple genes are involved in the regulation of toxin A yields. Classes 1 to 3 produced less than 1% of parental levels of extracellular toxin A. Class 1 mutants are apparently specific for toxin A. Class 2 mutants are pleotropic and produced toxin A, protease, and other extracellular proteins at reduced yields. Class 3 mutants are pleotropic and in addition have relatively high levels of cell-bound toxin A. Class 4 mutants produce toxin A at levels greater than 1% of parental yields. Of 16 toxin A-deficient mutants examined, only 1 was a class 1 mutant. This mutant (PA103-29) was shown to be identical to the parental strain in all respects tested except for its marked deficiency in toxin A. The suitability of this class 1 mutant for use in virulence studies is discussed.

Bacterial Proteins↗
PubMed1979

Effects of pseudomonas toxin A, diphtheria toxin, and cholera toxin on electrical characteristics of turtle bladder.

Rapidly developing changes in the short-circuiting current (Isc), conductance (G), and potential (PD) of turtle bladders in Na-rich or Na-free media are seen after the mucosal addition, at 10 nM, of each of three toxins that contain ADP-ribosylation activity: Pseudomonas aeruginosa toxin A, diphtheria toxin, and cholera toxin. Toxin A irreversibility decreased the Isc, PD, and G of bladders in Na-rich media and the Isc and PD of bladders in Na-free media. Diphtheria or cholera toxin reversibly increased Isc and PD (not G), but only in Na-free media. The effects of toxin A in the turtle bladder, like those in other host cell systems, were eliminated by preexposure of this toxin to heat, specific antitoxin, or dithiothreitol and urea. Because exposure to this last condition increases the ADP-ribosylation activity of toxin A, it is suggested that the proenzyme is the required transport-inhibiting form of toxin A. The effects of all three toxins occurred rapidly, possibly before any of the possible intracellular ADP-ribosylation reactions are initiated. Whereas a recognition binding of toxin of toxin to receptors on the apical membrane completely accounts for the reversible effects of diphtheria or cholera toxin, this and additional toxin-membrane interactions (e.g., translocation) are needed to account for the irreversible effects of toxin A.

Animals↗
PubMed1978

Effect of iron on yields of exotoxin A in cultures of Pseudomonas aeruginosa PA-103.

The yields of exotoxin A in Pseudomonas aeruginosa cultures were influenced by the concentration of iron in the culture media. When the iron concentration of the culture media was increased from 0.05 to 1.5 microgram/ml, there was at least a 90% decrease in exotoxin A (measured both by enzymatic activity and by mouse lethality) and a slight increase in the growth of the bacteria. The addition of iron as late as 13 h after initiation of growth repressed further measurable increases of exotoxin A within 3 h. Intracellular toxin levels were also reduced by increasing the iron concentrations of the culture media. The addition of 3.0 microgram of iron per ml did not significantly alter either the enzyme activity of preformed crude or purified exotoxin A or the mouse toxicity of the pure toxin. Thus it appears that either the rate of production or the rate of intracellular degradation of exotoxin A is regulated by the concentration of iron in the culture media.

Bacterial Toxins↗
PubMed1978

Isolation and characterization of a high-molecular-weight polysaccharide from the slime of Pseudomonas aeruginosa.

A procedure is described for isolating a high-molecular-weight polysaccharide (PS) from the slime of Pseudomonas aeruginosa immunotype 1. The resultant material, obtained from the void volume of a Sephadex G-100 column, was composed of carbohydrate and water. No lipopolysaccharide (LPS), 2-keto-3-deoxyoctonoate, heptose, phosphate, or protein was detectable, and nucleic acid contamination was generally below 1%. The carbohydrate composition of the PS was glucose, rhamnose, galactose, arabinose, and mannose. PS had a molecular weight of between 100,000 and 350,000 and did not disaggregate when chromatographed in the presence of sodium deoxycholate. An antigen immunologically indistinguishable from PS could be obtained from LPS by either acetic acid hydrolysis and column chromatography or by allowing solutions of LPS to stand at room temperature for 3 days. Some of this LPS-associated polysaccharide eluted as the void volume of a G-100 column but differed from PS by its lack of galactose and arabinose. LPS also contained an immunodeterminant not shared with PS that was detected by its stability to dilute alkali treatment (0.1 N NaOH, 37 degrees C, 2 h). PS was destroyed by alkali treatment. PS appeared to represent a form of LPS polysaccharide side chain that contains galactose and arabinose and is of a high molecular weight.

Antigens, Bacterial↗
PubMed1978

Protective immunity induced in mice by immunization with high-molecular-weight polysaccharide from Pseudomonas aeruginosa.

A high-molecular-weight alkali-labile polysaccharide (PS) isolated from the slime of immunotype 1 Pseudomonas aeruginosa was tested for its ability to protect mice from lethal challenge with the live, homologous organism. Intraperitoneal (i.p.) injection of 10 to 25 microgram of the PS protected 60 to 70% of the mice against challenge with up to 50 50% lethal dose units. Although single immunization of mice with up to 250 microgram of PS effected protective levels of only 70%, two successive immunizations provided 100% protection. Subcutaneous and intravenous immunization with PS also provided protection to i.p. challenges with immunotype 1 P. aeruginosa, but not to i.p. challenge with immunotype 4 P. aeruginosa. Although lipopolysaccharide (LPS) was found to be more immunogenic than PS in out studies, contamination of the alkali-labile PS with LPS did not account for the protection seen. Alkali treatment (0.1 N NaOH, 37 degrees C, 2 h) of the PS destroyed its protective effectiveness, while similarly treated LPS retained its capacity for inducing immunity in mice. Adsorption and passive protection studies with sera raised to either PS or a mixture of PS and LPS indicated that antibody directed to the alkali-labile PS antigen was capable of contributing to the protection of mice against challenge with P. aeruginosa.

Animals↗
PubMed1977

Pyocin sensitivity of Neisseria gonorrhoeae and its feasibility as an epidemiological tool.

Pyocin inhibition of Neisseria gonorrhoeae and its feasibility as a gonococcal typing scheme were examined. Mitomycin C-induced pyocin lysates of Pseudomonas aeruginosa were able to selectively inhibit the growth of gonococcal strains. The particles associated with the inhibitory activity were non-dialyzable, heat labile, Pronase sensitive, trypsin resistant, and of large molecular weight by membrane and gel filtration techniques. The inhibitory activity was shown to be specific by absorption with sensitive and insensitive strains of N. gonorrhoeae and P. aeruginosa. Partial purification of pyocin lysates by ammonium sulfate precipitation followed by ultracentrifugation revealed phagelike particles consistent with high-molecular-weight R-type pyocines. These particles were associated with increased inhibitory activity and could be seen associated with the gonococcal cell surface. One hundred and six gonococcal strains could be differentiated on the basis of their sensitivity of 23 pyocin extracts. Thirty different patterns of pyocin inhibition were seen. Isolates from different body sites from the same patient could generally be identified as being similar strains. Strains isolated from known consorts had the same patterns. In general, agreement between pyocin typing and available epidemiological information was good.

Bacteriocins↗
PubMed1977

Production of a capsule of Neisseria gonorrhoeae.

Neisseria gonorrhoeae can be induced to form a large capsule that can be visualized with India ink. Encapsulated gonococci appeared to be resistant to non-antibody-mediated phagocytosis compared with unencapsulated gonococci of the same strain and colony type.

Neisseria gonorrhoeae↗
PubMed1977

Incidence of exotoxin production by Pseudomonas species.

Pseudomonas aeruginosa exotoxin A has been shown to catalyze the transfer of the adenosine 5'-diphosphate (ADP)-ribose moiety of nicotinamide adenine dinucleotide onto elongation factor 2, resulting in the inhibition of mammalian protein synthesis. The enzymatic activity (ADP-ribosyl [ADPR]-transferase) is thought to account for the toxicity of exotoxin A. The distribution of the expression of exotoxin A within Pseudomonas species was examined. Laboratory strains as well as clinical isolates of Pseudomonas aeruginosa were tested. The production of exotoxin A was determined by assaying for ADPR-transferase activity in dialyzed frozen (-20 degrees C) and thawed cell-free supernatants from 22-h cultures or in 10-fold-concentrated supernatants. In addition, toxin production was detected immunologically using a modified Elek test. Exotoxin A production was detected in approximately 90% of the 111 isolates of P. aeruginosa. In contrast, none of the other species of Pseudomonas examined produced exotoxin A detectable by either ADPR-transferase activity or immunological reactivity.

Epitopes↗
PubMed1977

Variability of the lytic susceptibility of Neisseria gonorrhoeae to human sera.

The bactericidal activity of human sera for Neisseria gonorhoeae was studied. Sera were obtained from a group of patients with gonococcal infections who had acute urethritis, acute pelvic inflammatory disease, disseminated gonococcal infection, or who were asymptomatic carriers. The homologous and heterologous strains were tested with these sera. The development of serum bactericidal antibodies was not a universal event. With few exceptions, the susceptibility of a particular strain to human antibody and complement appeared to be largely independent of the particular person from whom the serum was obtained and was due instead to antigenic properties intrinsic to each individual strain. Lipopolysaccharide appeared to be the predominant antigen against which bactericidal antibodies were directed. The principal bactericidal antibody class was IgM. Blocking antibodies were not found to account for the lack of lytic activity. A correlation of bactericidal antibodies with protection from developing gonococcal infection could not be demonstrated in three pateints.

Antibodies, Bacterial↗