Analysis of parameters affecting the hemagglutination activity of Escherichia coli possessing colonization factor antigens: improved medium for observing erythrocyte agglutination

Sedlock, David M.; Bartus, H F; Zajac, Ian; Actor, Paul

doi:10.1128/jcm.13.2.301-308.1981

Cited by 9 publications

(8 citation statements)

References 26 publications

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“…The different purified P fimbriae did not readily agglutinate human erythrocytes. To improve hemagglutination, the low-ionic-strength buffer of Sedlock et al (30) was applied, but without success. However, after treatment of the erythrocytes with neuraminidase, the Pl and p2k erythrocytes became agglutinable by all four P fimbriae preparations.…”

Section: Resultsmentioning

confidence: 99%

P-antigen-recognizing fimbriae from human uropathogenic Escherichia coli strains

Korhonen¹,

Väisänen²,

Saxén³

et al. 1982

Infect Immun

151

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P-antigen-recognizing fimbriae (P fimbriae) from four pyelonephritogenic Escherichia coli strains and type 1 fimbriae from an E. coli strain and a Salmonella typhimurium strain were purified. The P fimbriae were morphologically similar to type 1 fimbriae. The purified P fimbriae agglutinated neuraminidase-treated human P1 and P2k erythrocytes but not p erythrocytes, which lack all P-blood group-specific glycosphingolipids. However, coating of neuraminidase-treated p erythrocytes with globoside rendered such erythrocytes agglutinable by the P fimbriae. The hemagglutinations were in all instances fully inhibited by the synthetic alpha-D-Galp-(1-4)-beta-D-Galp-1-O-Me glycoside. The binding specificity of the P fimbriae could also be demonstrated by using fimbriae coated onto latex particles and nontreated erythrocytes. It was thus concluded that the P fimbriae recognize and bind to the alpha-D-Galp-(1-4)-beta-D-Galp carbohydrate sequence occurring in the series of P-blood group antigen-specific glycosphingolipids. In contrast to both type 1 fimbriae, all four P fimbriae preparations showed multiple bands in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Antisera were raised in rabbits against the various E. coli fimbriae. In enzyme-linked immunosorbent assays each one of the antisera to the P fimbriae reacted to titers of log 4 to 7 with both the homologous and the heterologous P fimbriae, but not with the type 1 fimbriae of E. coli. In a reciprocal fashion, the antiserum to the type 1 fimbriae of one E. coli strain reacted only with the homologous type 1 but not with any of the P fimbriae preparations.

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Section: Resultsmentioning

confidence: 99%

P-antigen-recognizing fimbriae from human uropathogenic Escherichia coli strains

Korhonen¹,

Väisänen²,

Saxén³

et al. 1982

Infect Immun

151

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“…Since most bacterial attachment and hemagglutination reflect the microorganism's ability to overcome repulsive (negative) electrostatic surface changes (12), it is not surprising that Sedlock et al (29) have found that bacterial hemagglutination assays can be made more sensitive by lowering the pH and ionic strength of the buffering medium used. Failure to recognize and control the above-mentioned variables may account for some of the reported discrepancies in B. bronchiseptica's hemagglutinating ability.…”

Section: Sismentioning

confidence: 99%

Hemagglutination by Bordetella bronchiseptica

Bemis

Plotkin

1982

J Clin Microbiol

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A total of 53 isolates of Bordetella bronchiseptica from dogs and pigs were tested for their ability to agglutinate chicken, horse, sheep, dog, pig, and guinea pig erythrocytes. No differences in hemagglutinating activity were attributed to the animal origin of the bordetella isolates. Horse and dog erythrocytes consistently resulted in the strongest hemagglutination reactions, whereas only 4% of the B. bronchiseptica isolates produced weak agglutination of chicken erythrocytes. A total of 85% of the isolates agglutinated horse, sheep, dog, pig, and guinea pig erythrocytes. One canine isolate with hemagglutinating activity, strain 110H, was examined to determine the nature of the hemagglutinin(s) involved. Hemagglutination was always accompanied by hemadsorption, as determined by dark-field or phase-contrast microscopy. Treatment of cells and cell extracts with heat or protease K inhibited the hemagglutination reaction. Sonicated bacterial cells had a greater hemagglutinating ability than did unsonicated live bacteria. The hemagglutination reaction was not inhibited by any of 17 sugars nor by N - acetylglucosamine or ethylene glycol-bis-(β-aminoethyl ether)- N , N -tetraacetic acid. Hemagglutinins were not detected in sonic extracts nor in several bacterial subunit fractions, including isolated pili. Antigens in some of these preparations were, however, detectable by indirect hemagglutination with anti- B. bronchiseptica serum. Isolated pili could not be detected on the erythrocyte surface by electron microscopy; however, serial sections of erythrocytes agglutinated by the live Bordetella organisms showed that the bacterial outer membrane and the erythrocyte surface were separated by a space of approximately 20 nm. This study provided additional circumstantial evidence that B. bronchiseptica pili or at least heat-labile surface proteins which extend some distance from the bacterial surface are involved in hemagglutination. Multiple hemagglutinins are likely to exist within this species since one isolate lacking pili also agglutinated canine erthyrocytes. The hemagglutinins of B. bronchiseptica need to be isolated and characterized before the hemagglutination reaction can be applied to studies of attachment.

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“…Both human (Community Blood and Plasma, Folcroft, Pa.) and bovine erythrocytes (GIBCO Diagnostics, Madison, Wis.) were used in this study. The erythrocyte suspensions were prepared in a low-ionic-strength, isotonic buffer, the properties of which have been described elsewhere (14). The buffer consisted of 200 mM D-sorbitol, 50 mM D-mannose and 25 mM 2-(Nmorpholino)ethanesulfonic acid (Calbiochem, La Jolla, Calif.).…”

Section: Methodsmentioning

confidence: 99%

Use of a hemadsorption technique to evaluate the stability of the hemagglutination reaction of Escherichia coli cultures possessing human colonization factor antigens

Sedlock¹,

Bartus²,

Zajac³

et al. 1982

J Clin Microbiol

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Two strains of Escherichia coli, producing different colonization factor antigens (CFA), were monitored for the population density of CFA-producing bacteria after repeated subculture. The production of CFA was estimated by flooding agar plates containing isolated colonies with suspensions of human or bovine erythrocytes. The erythrocytes were suspended in a low-ionic-strength buffer and were fixed to CFA-positive colonies with a 1.0% tannic acid solution. Strain H-10407, possessing CFA/I fimbriae, showed a rapid loss of the hemagglutinin when subcultured, whereas strain CL-9699, producing CFA/II, was very stable. By using the hemadsorption assay, we could rapidly and easily distinguish CFApositive colonies from the CFA-negative variants. A survey of additional E. coli strains demonstrated the utility and specificity of the hemadsorption technique used.

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Analysis of parameters affecting the hemagglutination activity of Escherichia coli possessing colonization factor antigens: improved medium for observing erythrocyte agglutination

Cited by 9 publications

References 26 publications

P-antigen-recognizing fimbriae from human uropathogenic Escherichia coli strains

P-antigen-recognizing fimbriae from human uropathogenic Escherichia coli strains

Hemagglutination by Bordetella bronchiseptica

Use of a hemadsorption technique to evaluate the stability of the hemagglutination reaction of Escherichia coli cultures possessing human colonization factor antigens

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