Production of monoclonal antibodies against serotype strains of Pseudomonas aeruginosa

Lam, Joseph S.; MacDonald, Leslie A.; Lam, M. Y. C.

doi:10.1128/iai.55.11.2854-2856.1987

Cited by 27 publications

(11 citation statements)

References 11 publications

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“…Clinical strains of P. aeruginosa isolated from patients with CF and sera from patients were obtained from the State Serum Institute, Rigshospitalet, Copenhagen, Denmark. These strains were typed by using both the Pseudomonas typing sera (Difco Laboratories, Detroit, Mich.) and a set of 17 serotype-specific MAbs produced in our laboratory (16,17). A set of serial isolates, strains 49 to 58, was obtained from one patient with CF over a 1-month period, with samples being taken every 2 to 3 days.…”

Section: Methodsmentioning

confidence: 99%

Occurrence of a common lipopolysaccharide antigen in standard and clinical strains of Pseudomonas aeruginosa

et al. 1989

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The lipopolysaccharide (LPS) of Pseudomonas aeruginosa PAO1 contains two species of O polysaccharide termed A and B bands. The high-molecular-weight B-band LPS determines the O specificity of the bacterium, while the antigenically distinct A-band LPS consists of only shorter-chain polysaccharides. Seven hybridomas secreting A-band-specific monoclonal antibodies were produced and used to study the LPS of standard and clinical strains. Although A-band antibodies did not agglutinate any of the serotype strains presently in the International Antigenic Typing Scheme, Western immunoblots revealed that Il of the 17 serotype strains possessed A-band LPS. In a group of 250 clinical isolates from patients with cystic fibrosis, 170 (68%) had A-band LPS on the basis of agglutination tests, but in silver-stained gels all were shown to be deficient in O-antigen-containing B band. Investigation of serial isolates from a single patient revealed a pattern of antigenic variation. During the course of the infection, serotypeable isolates became nontypeable, and the O antigen was replaced with A band as the major LPS antigen. These results suggest that A-band LPS may be the major LPS antigen in nontypeable clinical isolates and a common antigen among other P. aeruginosa strains.

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Section: Methodsmentioning

confidence: 99%

Occurrence of a common lipopolysaccharide antigen in standard and clinical strains of Pseudomonas aeruginosa

et al. 1989

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“…In our laboratory, we use the IATS (149,212) designations for 20 reference serotype strains. MAb which differentiate between the first 17 of the 20 IATS serotypes have been generated (132,133), while the remaining 3 IATS serotypes can be distinguished by using adsorbed polyclonal sera. In a recent review (212), P. aeruginosa B-band LPS was divided into 31 chemotypes that include the 20 IATS serotypes and subtypes (i.e., strains that react with an MAb for a particular IATS serotype but have additional epitopes).…”

Section: Genetics Of B-band O-antigen Biosynthesis Identification Andmentioning

confidence: 99%

Genetics of O-Antigen Biosynthesis inPseudomonas aeruginosa

Rocchetta

Burrows

Lam

1999

Microbiol Mol Biol Rev

317

223

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SUMMARY Pathogenic bacteria produce an elaborate assortment of extracellular and cell-associated bacterial products that enable colonization and establishment of infection within a host. Lipopolysaccharide (LPS) molecules are cell surface factors that are typically known for their protective role against serum-mediated lysis and their endotoxic properties. The most heterogeneous portion of LPS is the O antigen or O polysaccharide, and it is this region which confers serum resistance to the organism. Pseudomonas aeruginosa is capable of concomitantly synthesizing two types of LPS referred to as A band and B band. The A-band LPS contains a conserved O polysaccharide region composed of d-rhamnose (homopolymer), while the B-band O-antigen (heteropolymer) structure varies among the 20 O serotypes of P. aeruginosa. The genes coding for the enzymes that direct the synthesis of these two O antigens are organized into two separate clusters situated at different chromosomal locations. In this review, we summarize the organization of these two gene clusters to discuss how A-band and B-band O antigens are synthesized and assembled by dedicated enzymes. Examples of unique proteins required for both A-band and B-band O-antigen synthesis and for the synthesis of both LPS and alginate are discussed. The recent identification of additional genes within the P. aeruginosa genome that are homologous to those in the A-band and B-band gene clusters are intriguing since some are able to influence O-antigen synthesis. These studies demonstrate that P. aeruginosa represents a unique model system, allowing studies of heteropolymeric and homopolymeric O-antigen synthesis, as well as permitting an examination of the interrelationship of the synthesis of LPS molecules and other virulence determinants.

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“…Various efforts have been made to serotype l? aeruginosa (5,12,21,24,25,39,43). Different serological typing schemes based on 0antigens describe from five ( 3 ) up to 25 0groups (45) of the bacteria, and a distinct 0serotype in one system may contain two or more type-specific epitopes in other typing systems (19,27).…”

Section: Discussionmentioning

confidence: 99%

Antibodies from chronically infected cystic fibrosis patients react with lipopolysaccharides extracted by new micromethods from all serotypes of Pseudomonas aeruginosa

Fomsgaard

Shand

Freudenberg

et al. 1993

APMIS

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N. Antibodies from chronically infected cystic fibrosis patients react with lipopolysaccharides extracted by new micromethods from all serotypes of Pseudomonas aeruginosa. APMIS 101: 101-1 12, 1993. Micromethods were developed to extract lipopolysaccharide (LPS, endotoxin) from single bacterial colonies of the 20 recognized Pseudomonas aeruginosa type strains. The appearance of these LPSs in polyacrylamide gel electrophoresis (PAGE) and their reactivity with serum ofcystic fibrosis (CF) patients chronically infected with f! aeruginosa was studied. Silver staining of LPS after PAGE showed that 13 of the R aeruginosa LPSs had high numbers of 0-repeating units arranged in 1 4 clusters of banding. Low-molecular-weight LPS fractions were more prominent in six of the serotype strains, of which 0:7 and 0: 14 appeared semi-rough. Corresponding immunoblots using the CF sera showed LPS patterns very similar to the silver-stained appearance, indicating an immune reaction to all P aeruginosa LPS including that from the newly discovered 0:18, 0:19 and 0:20. This was unexpected since only a few serotype strains (mostly 0:3,0:6 and 0:9) had been isolated from the patients. Absorption experiments using purified and chemically defined P aeruginosa rough LPS and smooth LPS suggested these immune reactions were due to antibodies cross-reactive to corellipid A as well as to lower molecular weight 0polysaccharides or "A-bands". However, in some cases 0:3, 0 : 6 , and 0:9 LPSs were also found to contain additional distinct 0-epitopes. Immune recognition of various polyagglutinable II aeruginosa LPSs seemed also to be caused by cross-reactive antibodies. The described microextraction methods, followed by PAGE and silver staining or immunoblotting, are easy and convenient techniques with which to study antibodies against LPS epitopes and to screen for LPS phenotypic appearance using only a few bacterial colonies from larger numbers of Gram-negative bacterial strains.

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Production of monoclonal antibodies against serotype strains of Pseudomonas aeruginosa

Cited by 27 publications

References 11 publications

Occurrence of a common lipopolysaccharide antigen in standard and clinical strains of Pseudomonas aeruginosa

Occurrence of a common lipopolysaccharide antigen in standard and clinical strains of Pseudomonas aeruginosa

Genetics of O-Antigen Biosynthesis inPseudomonas aeruginosa

Antibodies from chronically infected cystic fibrosis patients react with lipopolysaccharides extracted by new micromethods from all serotypes of Pseudomonas aeruginosa

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