Biomedical subjects

G B Pier

Publications and source records attributed to G B Pier.

At least 145 records · Page 8Linked to original sources

PubMed1983

Characterization of the human immune response to a polysaccharide vaccine from Pseudomonas aeruginosa.

Sera from humans vaccinated with a high-molecular-weight polysaccharide vaccine to Pseudomonas aeruginosa immunotype 1 (IT-1) were analyzed for duration of the immune response, specificity for the IT-1 determinant, and by assessing the immunoglobulin classes elicited. The ability of purified IgG, IgM, and IgA to interact with peripheral blood leukocytes, as well as purified polymorphonuclear neutrophils or mononuclear cells, was also examined in an opsonophagocytosis assay. Levels of antibody to IT-1 remained significantly (P less than 0.001) elevated 21 months after immunization. Responses were generally specific to the IT-1 serotype determinant. Some vaccinees also responded to immunotype 2 and immunotype 5 determinants. IgG, IgM, and IgA serum antibodies were all elicited by vaccination. IgG and IgA were effective opsonins for P aeruginosa. IgM-mediated opsonophagocytosis required complement. Serum IgA was highly effective in conjunction with mononuclear cells in opsonophagocytosis of P aeruginosa, suggesting that these immune components may be capable of protecting neutropenic hosts.

Adult↗

PubMed1983

Safety and immunogenicity of a group 29E meningococcal capsular polysaccharide vaccine in adults.

A serogroup 29E Neisseria meningitidis capsular polysaccharide vaccine at a dose of 50 micrograms was injected subcutaneously into 10 adult human volunteers. One vaccinate experienced a mild systemic reaction; one complained of moderate to severe local pain and tenderness. The vaccine induced significant homologous binding and bactericidal antibody by 2 weeks and significant binding antibody against the heterologous serogroup Z capsular polysaccharide by 4 weeks. Although binding antibody rose during the first 4 weeks and then declined slowly over the subsequent 4 months, bactericidal antibody response declined substantially by 4 and 8 weeks for both polysaccharides. The increase in group 29E bactericidal activity was no longer significant at 4 and 8 weeks; loss of bactericidal activity against group Z was significant by 8 weeks. Bactericidal activity again rose between 8 and 26 weeks, becoming significantly increased over prevaccination levels for group 29E and increased, though not significantly (P = 0.085), over prevaccination levels for group Z. As a result of the failure of induced antibody to increase bactericidal activity, only 40% of vaccinates achieved a greater than 2 log2 increase in lytic activity against group 29E, and none achieved this large an increase in lytic activity against group Z.

Adolescent↗

PubMed1983

Characterization of the antibody response in inbred mice to a high-molecular-weight polysaccharide from Pseudomonas aeruginosa immunotype 1.

We explored the genetic basis for the differing immune responses observed in inbred strains of mice to a high-molecular-weight polysaccharide (PS) from Pseudomonas aeruginosa immunotype 1 (IT-1). Previous studies have shown that C3H mice immunized with this antigen produce only immunotype-specific antibody. BALB/c mice immunized with IT-1 PS produce both anti-IT-1 PS antibody and antibody cross-reactive with PS from P. aeruginosa immunotype 2 (IT-2). In the current study, we observed that, in addition, these two strains differ in their ability to respond to low immunizing doses of IT-1 PS. C3H mice generated a protective antibody response after a 1-microgram immunization, whereas BALB/c mice failed to produce protective antibody after receiving 1 microgram of PS. Both strains generated protective levels of antibody after a 50-micrograms immunization. Genetic analysis of these response patterns indicates that the ability to produce cross-reactive antibody and the ability to respond to a 1-microgram immunization are independently inherited traits. In addition, the responsiveness of C3H mice to a 1-microgram immunization with the production of protective levels of antibody is not linked to the mouse major histocompatibility (H-2) complex, to sex-linked genes, or to a single gene outside the H-2 complex.

Animals↗

PubMed1983

Pseudomonas aeruginosa isolates from patients with cystic fibrosis: a class of serum-sensitive, nontypable strains deficient in lipopolysaccharide O side chains.

Twenty-six Pseudomonas aeruginosa strains from patients with cystic fibrosis were typed by the Fisher immunotyping scheme. Only 6 strains were agglutinated by a single typing serum, whereas 15 strains were agglutinated with more than one serum and 5 were not agglutinated by any serum. Neither the polyagglutinable nor the nonagglutinable strains were typable by hemagglutination inhibition or immunodiffusion, suggesting that these polyagglutinable strains did not express multiple serotype antigens, but were instead being agglutinated by antibody to nonserotype determinants. Four typable isolates were resistant to pooled normal human serum, whereas the 12 polyagglutinable and nonagglutinable isolates studied were very sensitive to normal human serum. The outer membranes of 16 strains were isolated and characterized. The data suggested, in general, strong conservation of outer membrane protein patterns. Lipopolysaccharides (LPS) were purified by a new technique which allowed isolation of both rough and smooth LPS in high yields. Three of four typable, serum-resistant strains examined had amounts of smooth, O-antigen-containing LPS equivalent to our laboratory wild type, P. aeruginosa PAO1 strain H103. In contrast, 10 of 12 polyagglutinable or nonagglutinable, serum-sensitive strains had very little or no smooth, O-antigen-containing LPS, and the other two contained less smooth LPS than our wild-type strain H103. In agreement with this data, five independent, rough, LPS O-antigen-deficient mutants of strain H103 were nontypable and serum sensitive. We suggest that the LPS defects described here represent a significant new property of many P. aeruginosa strains associated with cystic fibrosis.

Bacterial Outer Membrane Proteins↗

PubMed1983

Further purification and characterization of high-molecular-weight polysaccharide from Pseudomonas aeruginosa.

Previously published reports on high-molecular-weight polysaccharides from immunotype 1 and 2 of Pseudomonas aeruginosa indicated the presence of high levels of mannose in these preparations. This mannose has been found to be due to the presence of a yeastlike mannan in high-molecular-weight polysaccharide preparations. The source of the mannan was found to be the tryptic soy broth used to grow the bacteria. Mannan could be removed from the polysaccharide preparations by chromatography over columns of concanavalin A-Sepharose. The resulting polysaccharides had the same serological reactivity against rabbit antisera and the same immunogenic properties in mice as did the mannan-containing polysaccharides. Comparison of mannan-depleted polysaccharide with preparations of high-molecular-weight polysaccharide obtained from either ultrafiltered tryptic soy broth or a chemically defined medium showed that these polysaccharides were immunologically and chemically similar. Human immune responses to mannan-depleted polysaccharide from the immunotype 1 strain of P. aeruginosa were comparable with those previously seen in humans receiving mannan-containing polysaccharides. Thus, we found that P. aeruginosa high-molecular-weight polysaccharides prepared in either tryptic soy broth and then subjected to concanavalin A-Sepharose chromatography, ultrafiltered tryptic soy broth, or a chemically defined medium were immunologically and chemically comparable.

Animals↗

PubMed1982

Lipopolysaccharide and high-molecular-weight polysaccharide serotypes of Pseudomonas aeruginosa.

The serotype distribution of bacteremic and nonbacteremic clinical isolates of Pseudomonas aeruginosa in relation to the Fisher immunotyping scheme, the International Antigenic Typing System (IATS), and high-molecular-weight polysaccharide determinants was investigated. Of 281 bacteremic isolates, 273 (97.2%) were serotyped by one of the seven IATS specificities that correspond to a Fisher lipopolysaccharide/high-molecular-weight polysaccharide specificity. In contrast, these seven serotypes accounted for only 68.5% of the 124 nonbacteremic clinical isolates. Review of the reported serotype distribution of P. aeruginosa isolates in Europe further supported the finding of a limited serotype distribution among bacteremic clinical isolates. Fifteen of the 17 IATS serotypes were found among all of the strains of P. aeruginosa serotyped, an indication that most of the IATS serotypes are present in the United States. Thus, only certain lipopolysaccharide immunotypes of P. aeruginosa occur as clinical bacteremic isolates, and a multivalent, high-molecular-weight polysaccharide vaccine directed at the lipopolysaccharide type determinants of P. aeruginosa has potential usefulness.

Agglutination Tests↗

PubMed1982

Immunological basis of serum resistance of Neisseria gonorrhoeae.

The immunological basis for resistance of certain strains of Neisseria gonorrhoeae to the bactericidal action of normal human serum was studied by investigating the potential role of factors which are known to interfere with each of the sequential steps that result in immune lysis of Gram-negative bacteria. Strains of N. gonorrhoeae were characterized as serum-sensitive (sers) or serum-resistant (serr) on the basis of their sensitivity to lysis by the sera of six normal individuals. Neither intrinsic resistance to the lytic action of activated human complement nor inaccessibility of the cell membrane to C5b accounted for serr. Outer membrane lipopolysaccharide (LPS) was the target antigen for lytic antibody in normal human sera. The gross chemical composition and molecular size of the LPS of the strains were heterogeneous and no consistent patterns of differences between those extracted from serr and from sers strains were found. Neither IgA nor IgG 'blocking' antibody in normal human serum was responsible for serr. We conclude that serr results from the absence from the LPS of the strains of antigenic loci for the lytic antibody in most normal human sera, or, expressed as a function of the host, the absence from the sera of most normal humans of lytic antibody directed against LPS antigenic loci for immune lysis.

Antibodies, Bacterial↗

PubMed1982

Cross-protection by Pseudomonas aeruginosa polysaccharides.

High-molecular-weight polysaccharide from Pseudomonas aeruginosa immunotypes 1 and 2 gave cross-protection in outbred CD-1 mice challenged with the heterologous immunotype organism. Both active immunization with 50 micrograms of polysaccharide, as well as passive transfer of immune serum were effective. The basis for this cross-protection is the ability of high doses of polysaccharide to induce antibody formation to both homologous and heterologous immunotype determinants.

Animals↗

PubMed1982

Safety and immunogenicity of high molecular weight polysaccharide vaccine from immunotype 1 Pseudomonas aeruginosa.

The safety and immunogenicity of a high molecular weight polysaccharide from immunotype 1 Pseudomonas aeruginosa were tested in a dose response fashion in adult volunteers. The vaccine lacked toxicity and pyrogenicity for experimental animals. Doses of 50, 75, 150, or 250 microgram were given to groups of individuals as a single dose subcutaneous injection. Doses of 150 and 250 microgram were associated with a significant rise in binding and opsonic antibody at 2 wk postimmunization. Titers remained unchanged for up to 6 mo. The vaccine was almost devoid of toxicity, eliciting no more than a slightly sore and tender arm at the site of injection. High molecular weight polysaccharide antigen appears to induce a good immune response following vaccination that is effective in mediating opsonophagocytic killing of live P. aeruginosa organisms.

Animals↗

PubMed1982

Induction in mice of cell-mediated immunity to Pseudomonas aeruginosa by high molecular weight polysaccharide and vinblastine.

The effect of the cytotoxic drug vinblastine on the development of immunity to high m.w. polysaccharide (PS) isolated from culture supernates of Pseudomonas aeruginosa was investigated. One microgram of PS, a normally nonimmunogenic, nonprotective dose, plus 75 micrograms of vinblastine were administered to BALB/c mice, and afforded protection to live organism challenge with the homologous strain. The kinetics and serotype specificity of the immune response indicated an active immunization had occurred. Analyses of serum antibody levels of mice given the PS-drug regimen in a sensitive, radioactive antigen-binding assay (RABA) failed to show development of antibody to the immunizing PS. Immunity could be passively transferred with spleen cells but not by serum from PS-drug-immunized animals, and the effector cell was removed by antisera to the Thy-1.2 antigen. Nu/nu mice were also protected against challenge after immunization with PS and vinblastine, but this protection was observed in association with the development of serum antibody to PS in these mice, as measured in the RABA. Protective immunity could not be elicited in the BALB/c mice by PS plus cyclophosphamide. These data suggest that under certain conditions, PS antigens can elicit T cell-dependent immune phenomena, and this T cell-dependent immunity can protect mice from live organism challenge against an extracellular bacterial pathogen.

Animals↗

PubMed1981

Mucoid Escherichia coli in cystic fibrosis.

Patients with cystic fibrosis commonly harbor in their lungs strains of Pseudomonas aeruginosa that have a mucoid coating considered virtually pathognomonic for the disease. We found that strains of Escherichia coli with a morphologically similar mucoid coating were present in the respiratory tracts of eight (11.8 per cent) of 68 patients with cystic fibrosis whose sputum cultures yielded Esch. coli, as compared with none of 89 patients without cystic fibrosis who had Esch. coli in sputum. Mucoid strains of Esch. coli were also recovered from the stools of five (11.1 per cent) of 45 patients with cystic fibrosis, as compared with one (0.7 per cent) of 150 patients without cystic fibrosis. The mucoid substances purified from Esch. coli were biochemically and antigenically distinct from those of P. aeruginosa. We conclude that the respiratory tract in cystic fibrosis offers an environment conducive to the production of a mucoid coating not only by P. aeruginosa but by other gram-negative bacilli as well.

Bacteriological Techniques↗

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

Safety and immunogenicity of group Y and group W135 meningococcal capsular polysaccharide vaccines in adults.

Serogroup Y and W135 Neisseria meningitidis capsular polysaccharide vaccines were tested as monovalent and divalent preparations in groups of 10 adult human volunteers at a dose of 50 (monovalent) or 100 micrograms (divalent) injected subcutaneously. Reactogenicity was low for the group Y vaccine and the group Y-W135 combined vaccine; 3 of 10 volunteers developed systemic reactions after group W135 vaccination. All three vaccines induced significant homologous and heterologous binding and bactericidal antibody. Except for group W135 bactericidal antibody, homologous responses exceeded heterologous responses, and divalent and monovalent vaccines induced equivalent homologous responses. Homologous bactericidal antibody responses were maintained for 4 weeks in 85% of group W135 vaccinates and in 100% of group Y vaccinates. Bactericidal antibody was induced in 11 of 11 group Y and 12 of 15 group W135 volunteers without preexisting respective bactericidal activities, regardless of which vaccine they received. For all three vaccines, antibody levels declined only slightly over 6 months. Prevaccination antibody levels positively affected postvaccination binding antibody levels, but not bactericidal levels.

Antibodies, Bacterial↗

PubMed1981

Correlation of the biologic responses of C3H/HEJ mice to endotoxin with the chemical and structural properties of the lipopolysaccharides from Pseudomonas aeruginosa and Escherichia coli.

The basis of the biologic responses of C3H/HeJ mice to endotoxin administration in relation to the structural linkages in the lipid A portion of the lipopolysaccharide (LPS) of Pseudomonas aeruginosa and Escherichia coli were investigated. P. aeruginosa LPS was found to be immunogenic, mitogenic, and toxic, but not lethal, in C3H/HeJ mice. The observed mitogenicity in spleen cells was directed toward immunoglobulin- (Ig) bearing cells, was present in response to isolated and solubilized lipid A, and was inhibitable by polymixin B. The P. aeruginosa LPS was chemically analyzed in order to define its composition and exclude the presence of contaminating proteins being responsible for the biologic responses of C3H/HeJ mice that were observed. Structural analysis of the linkages of the fatty acids to the glucosamine backbone in the lipid A of P. aeruginosa and E. coli revealed similarities in terms of the ratio of hydroxy fatty acids to straight chain fatty acids and the way in which these 2 types of fatty acids were linked to the backbone. Differences were seen in the carbon chain length of the fatty acid substituents, and the substituent on the hydroxy fatty acid that is directly ester linked to the glucosamine backbone. These data indicate that the refractivity of C3H/HeJ mice to the biologic effects after the administration of Gram-negative endotoxins may be limited to enterobacterial LPS. Those differences we found in the chain length and/or linkages of the fatty acid substituents in the lipid A portion of the LPS between P. aeruginosa and E. coli may be sufficient to render C3H/HeJ mice responsive to the biologic effects of nonenterobacterial endotoxins.

Animals↗

PubMed1980

Elaboration of both the group W135 and group Y capsular polysccharides by a single strain of Neisseria meningitidis.

A single strain (8021) of Neisseria meningitidis, isolated from a child with disseminated meningococcal disease, was found to elaborate two serogroup-specific capsular polysaccharides-Y and W135. The original isolate as well as the progeny of ten single colony sub-isolates each agglutinated with both group Y and group W135 serogrouping antisera. The capsular polysccharide of strain 8021 contained the chemical constituents of both the W135 and Y capsular polysaccharides in a ratio of about 2.5:1. The patient responded immunologically to both capsular polysaccharides with haemagglutinating antibodies. Analysis by double diffusion in agar revealed that the capsular polysaccharide of strain 8021 contained individual molecules of group W135 and group Y capsular polysaccharides as well as a mosaic molecule containing both antigenic determinants.

Antigens, Bacterial↗

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↗

PubMedDate not supplied

[Pseudomonas aeruginosa immunoglobulin in experimental pneumonia].

A guinea pig model of experimental Pseudomonas aeruginosa pneumonia was used to evaluate factors affecting the efficacy of passive immune therapy with Psomaglobin N, a hyperimmune P. aeruginosa globulin. Animals treated 2 h after infection with a single intravenous infusion of Psomaglobin N, 500 mg/kg, demonstrated 33% survival. Lower dosages were less effective and no survivors occurred among albumin-treated controls. Treatment with Psomaglobin N was effective if given 2 h or 8 h after infection but not if delayed until 24 h after infection. Animals rendered neutropenic with cyclophosphamide did not survive if treated with Psomaglobin N alone. However, when Psomaglobin N was added to tobramycin treatment, a significant increase in survival occurred (86%) as compared to that observed with tobramycin alone (43%) (p less than 0.05). We conclude that Psomaglobin N may offer a useful therapeutic option in management of P. aeruginosa pneumonia.

Animals↗