Avian P1 Antigens Inhibit Agglutination Mediated by P Fimbriae of Uropathogenic Escherichia coli
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INFECTION AND IMMUNITY, Feb. 1992, p. 578-583 Vol. 60, No. 2
0019-9567/92/020578-06$02.00/0
Copyright © 1992, American Society for Microbiology
Avian P1 Antigens Inhibit Agglutination Mediated by P Fimbriae of
Uropathogenic Escherichia coli
JAMES R. JOHNSON,'* JANE L. SWANSON,2 AND MARGUERITE A. NEILL3
Departments of Medicine' and Laboratory Medicine and Pathology,2 University of Minnesota, Minneapolis,
Minnesota 55455, and Department of Medicine, Brown University, Providence, Rhode Island 029123
Received 26 August 1991/Accepted 18 November 1991
Whole egg white from pigeon, dove, and cockatiel eggs, as well as the ovomucoid fraction of pigeon egg white,
exhibited strong P1 antigenic activities and inhibited agglutination of human P1 erythrocytes and of
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digalactoside-coated latex beads by P-fimbriated Escherichia coli strains. In contrast, chicken egg white
exhibited only weak P1 antigenic activity and had little impact on P-fimbrial agglutination. These preparations
did not affect hemagglutination by E. coli strains expressing mannose-resistant adhesins other than P fimbriae,
i.e., Dr, F1845, and S adhesins. Human anti-P1 serum diminished the P-fimbrial inhibitory activities of pigeon
egg white and pigeon ovomucoid. Pigeon ovomucoid was equipotent on a molar basis with globoside, and the
pigeon, dove, and cockatiel egg white preparations were equipotent with each other in P-fimbrial inhibition.
Incubation of p erythrocytes in whole egg whites or in pigeon ovomucoid did not render them agglutinable by
P-fimbriated bacteria, whereas incubation in globoside did. These data demonstrate that whole egg whites (and
their ovomucoid fraction) from members of the families Columbidae (pigeons and doves) and Psittacidae
(parrots) specifically and potently inhibit P-fimbrial agglutination, probably by providing P1 antigen as a
receptor for the P-fimbrial adhesin. Avian egg white preparations may facilitate adhesin characterization of
wild-type uropathogenic strains and may be useful in preventing upper urinary tract infections due to
P-fimbriated E. coli.
Pyelonephritogenic Escherichia coli strains are able to P1 antigenic activity has been identified in the egg white,
colonize the upper urinary tract (29) and to cause pyelone- blood, and/or excreta of members of two avian families,
phritis (12) in part because of their expression of P fimbriae. Columbidae (i.e., pigeons [2, 11, 30] and doves [6, 7]) and
P fimbriae are so named because they bind to the digalacto- Psittacidae (i.e., budgerigars [2, 6]), but has been reported to
side-containing antigens of the human P blood group system, be absent from the tissues and products of other birds (2, 6).
such as the P1 antigen (5, 14, 16, 17, 20, 21, 25). Digalacto- We sought to determine whether P1-antigen-containing egg
side-containing glycolipids and oligosaccharides have been whites from members of the Columbidae and Psittacidae
used to block P-fimbrial binding in the laboratory character- families could serve as inhibitors of P-fimbrial adherence and
ization of adhesin specificity among uropathogenic E. coli to characterize the specificity, potency, and mechanism of
strains (14, 16, 17, 20, 21) and in the prevention of experi- such inhibition. We also sought to discover whether P1-
mental upper urinary tract infection due to P-fimbriated E. antigen-containing egg white preparations share with globo-
coli (15, 31, 32). Such compounds are prohibitively expen- side the property of attaching to human cells and increasing
sive and thus are impractical for general use. An additional the density of receptors for P fimbriae.
problem in their use to prevent pyelonephritis is that gly-
colipids such as globoside can insert into host cell mem-
branes and can thereby increase the P-fimbrial receptor MATERIALS AND METHODS
density and potentially promote infection rather than pre- Plasmids and strains used. Two strains expressing cloned
vent it (9). P-fimbrial determinants from strain J96, i.e., HU849 (29) and
Alternative approaches to the adhesin characterization of JJ48, were used as P-fimbrial controls. To obtain P-fimbrial
uropathogenic E. coli include agglutination tests with diga- expression greater than that found with strain HU849, strain
lactoside-coated latex beads (4) or human erythrocytes that JJ48 was created by subcloning the P-fimbrial gene cluster
express P blood group system antigens (i.e., P1, P2, pk, and from pRHU845 (26) into pUC19 (35) at the EcoRI site and
P2 erythrocytes) or do not express P antigens (i.e., p transforming this construct (pJJ48) into host strain DH5-ao by
erythrocytes) (5, 14, 16, 17, 20, 25). However, latex bead standard techniques (24).
agglutination combined with hemagglutination of P+ eryth- E. coli strains expressing MR adhesins other than P
rocytes does not allow the identification of mannose-resis- fimbriae (non-P mannose-resistant [NPMR] adhesins) were
tant (MR) adhesins other than P fimbriae when P fimbriae are used to evaluate the specificity of agglutination inhibition by
also present. Furthermore, the limited availability and re- egg white preparations. Dr hemagglutinin and S fimbriae, the
stricted shelf life of p erythrocytes limit their use to special- two major NPMR adhesins of uropathogenic E. coli (12),
ized laboratories. Thus, an inexpensive inhibitor that specif- were each represented by a wild-type strain expressing only
ically blocks P-fimbrial binding and does not attach to host the NPMR adhesin of interest; a recombinant sfa+ strain
cells would be useful both for adhesin characterization and was also used for S fimbriae (Tables 1 and 2). Strain
for prevention of pyelonephritis.
HU968-115, which was isolated from the urine of a patient in
Houston, Tex., expresses sfa determinants and lacks pap
determinants (lOa). An F1845-fimbriated recombinant strain
*
Corresponding author. was included (Tables 1 and 2), since F1845 fimbriae are
578VOL. 60, 1992 AVIAN PI ANTIGENS INHIBIT P-FIMBRIAL AGGLUTINATION 579
TABLE 1. Plasmids used in this study
Plasmid Genotype Selectiona
replicatorand Reference
pRHU845 pap' recombinant plasmid (from pyelonephritis isolate J96) Tetr, pACYC184 10, 26, 29
pJJ48 pap+ recombinant plasmid (pap region from pRHU845 in pUC19) Ampr, pUC19 This study
pSSS1 F1845+ recombinant plasmid (from diarrheal isolate C1845) Ampr, pUC8 1
pJFK102 pap-2+ recombinant plasmid (from pyelonephritis isolate J96) Ampr, pBR322 18, 19
pANN801-13 sfa+ recombinant plasmid (from uroisolate 536) Ampr, pBR322 8
a
The resistance phenotype which was selected for is given.
closely related to the Dr hemagglutinin (28, 33) but recognize Preparation of inhibitors. The P antigen, containing an
a different portion of the Dr blood group antigen (28). The internal digalactoside moiety, is present on human cells as
F1845-fimbrial plasmid pSSS1 (2) was transformed into the glycosphingolipid globoside (25). Globoside (estimated
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P-fimbrial recombinant strain HU849 to create a strain molecular weight, 1,339) in chloroform (Matreya, Inc.,
expressing both P and NPMR adhesins (strain JJ105). Pleasant Gap, Pa.) was evaporated under a stream of nitro-
A recombinant strain expressing pap-2-encoded fimbriae gen and resuspended in PBS to give an estimated concentra-
(18, 19), which are a variant of P fimbriae with binding tion of 2.5 mM (3.3 mg/ml). This globoside stock solution
specificities for SSEA-4 and the Luke (LKE) human blood was sonicated briefly to form micelles (20) and stored at 4°C.
group antigen (extended derivatives of globoside), was also Chicken eggs were purchased from a supermarket. Cock-
included. This adhesin was first identified in a uropathogenic atiel, dove, and racing (homing) pigeon eggs were gifts from
strain and has been variously reported to mediate (19) or not local bird owners. Chicken egg white (CEW), cockatiel egg
to mediate (23) MR hemagglutination of human erythro- white (CLEW), dove egg white (DEW), and racing (homing)
cytes. HB101 (24), a nonfimbriated and MR-adhesin-nega- pigeon egg white (PEW) were aspirated from eggs, homog-
tive E. coli K-12 derivative (3), was used as a negative enized in a blender, sonicated for 3 h to permit microfiltra-
control in agglutination studies. tion, passed through a 0.2-,um-pore-size filter (Nalge/Sybron
Erythrocytes and P-antigen-coated latex beads. Erythro- Corp., Rochester, N.Y.), and stored at -20°C. Working
cytes obtained from one of the authors (J.R.J.) were con- stocks were kept at 40C.
firmed as type P1 by agglutination with human anti-P1 serum Racing pigeon ovomucoid (POM) was purified from whole
(Amtec Diagnostics International, Inc., Conroe, Tex.) as egg white according to the method of Lineweaver and
described by the manufacturer. Human P2, Pk, and p Murray (22). Briefly, whole egg white was extracted with 0.5
erythrocytes from storage in liquid N2 were obtained from M trichloroacetic acid-acetone (1:2, vol/vol) at 40C and then
the University of Minnesota Blood Bank and from John precipitated with acetone. The precipitate was dialyzed
Moulds (Gamma Biologicals, Inc., Houston, Tex.). Eryth- extensively against distilled water and passed through a
rocytes were prepared as a 5% (vol/vol) suspension in P-100 gel filtration column (Bio-Rad Laboratories, Rich-
phosphate-buffered saline (PBS) (Dulbecco's PBS without mond, Calif.) to separate POM from residual nonprotein
calcium or magnesium; Celox Corp., Minnetonka, Minn. species. Fractions containing P1 activity were pooled, dia-
[pH 7.1]) plus 3 or 10% (wt/vol) alpha-D-mannose for bacte- lyzed, and lyophilized. Reduction with 3-mercaptoethanol
rial agglutination reactions. Latex beads coated with the (by incubating purified native POM in 0.3% P-mercaptoeth-
Gal(al-4)Gal minimal receptor for P fimbriae (P beads) anol for 1 h and adding 0.3% ,B-mercaptoethanol to the
(Latex-Pl; Chembiomed, Edmonton, Alberta, Canada) were running buffer of the gel filtration column) resulted in a
used to detect expression of digalactoside-specific adhesins 10-fold increase in the P1 antigenic activity of the purified
(4, 13). The digalactoside moiety on P beads is chemically POM. As little as 100 ng of purified reduced POM inhibited
identical with the terminal digalactoside portion of the P1 P1 erythrocyte agglutination by high-titer anti-P1 serum.
antigen (25). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis
TABLE 2. Bacterial strains used in this study
Strain Recombinant MR-adhesin Selectionb Reference
plasmid content phenotype' or source
HU849 pRHU845 P Tetr 26, 29
JJ48 pJJ48 P Ampr This study
HB101/pSSS1 pSSS1 F1845 Ampr 1
JJ105 pRHU845, pSSS1 P, F1845 Tetr Ampr This study
IH11128 Nonec Dr None 27
P678-54/pJFK102 pJFK102 pap-2 Ampr 18, 19
HU968-115 None' S None This study
536-21 pANN801-13 S Ampr 8
DH5-at Noned None None BRLe
HB101 Noned None None 3
a p, P fimbriae; F1845, F1845 fimbriae; pap-2, pap-2-encoded fimbriae; S, S fimbriae; Dr, the Dr hemagglutinin.
b The resistance phenotype which was selected for is given.
Wild-type uroisolate.
d E. coli K-12 or K-12 derivative.
BRL, Bethesda Research Laboratories, Gaithersburg, Md.580 JOHNSON ET AL. INFECT. IMMUN.
of both native and reduced POM followed by Coomassie TABLE 3. Agglutination and agglutination inhibition patterns of
R-250 staining showed two bands migrating at 48 and 52 kDa. bacterial strains with PBS, CEW, CLEW, DEW, PEW, POM,
Western blot (immunoblot) showed that both bands bound and globoside
purified Shiga-like toxin I, which is known to bind via its Agglutination patteMa of MR adhesins
subunit to the Gal(otl-4)Gal linkage present in human P1 Type of agglutination and
inhibitor pb NPMR P plus NPMR
antigen. adhesinsc adhesinsd
Agglutination reactions. Bacterial suspensions (approxi-
mately 5 x 1010 CFU/ml) were prepared in PBS from P bead agglutination
PBS + - +
plate-grown bacteria (Luria agar, supplemented with antibi- CEW + - +
otics as necessary) (24) after incubation overnight at 37°C. CLEW - - -
Agglutination reactions were performed by combining 5 DEW - - _
volumes of bacterial suspension, 5 volumes of inhibitor PEW - - _
solution (or PBS), and 2 volumes of substrate (p beads or POM - - -
erythrocytes) on microscope slides over ice (S-fimbrial Globoside - -
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strains) or at room temperature (other strains). The intensity MR hemagglutination
of agglutination was scored on a scale from negative (-) to of P1 erythrocytes
most intense (+ + +). Serial twofold dilutions of each bacte- PBS + + +
rial suspension were tested for agglutination of erythrocytes CEW + + +
and P beads to determine the minimal agglutinating concen- CLEW - + +
tration (MAC [lowest bacterial concentration giving macro- DEW - + +
PEW - + +
scopic agglutination]) for each strain with each substrate. POM - + +
Agglutination inhibition experiments with each strain were Globoside - + +
performed at bacterial concentrations of four times the
strain's MAC. Inhibitors that completely blocked P-fimbrial MR hemagglutination of - + +
agglutination when tested undiluted were tested at serial p erythrocytes
twofold dilutions to determine the MICs for hemagglutina- " +, any agglutination (+, +, ++, or +++); -, agglutination absent even
tion and P bead agglutination by strain JJ48 (at four times its with undiluted bacterial suspension or with use of undiluted inhibitor solu-
MACs for these substrates). For each inhibitor, the lowest tions.
b p, P fimbriae (strains JJ48 and HU849).
concentration or dilution producing complete agglutination ' Similar results were obtained with F1845 fimbriae (strain HB101/pSSS1),
inhibition was designated the MIC10. The first dilution at the Dr hemagglutination (strain IH11128), and S fimbriae (strains HU968-115
which inhibition was no longer evident was designated the and 536-21).
d P fimbriae plus NPMR fibriae (strain JJ105).
MICo. The dilution giving 50% inhibition of agglutination (as
calculated by the geometric mean of the MIC10o and the
MICO) was designated the MIC50. Inhibitor concentrations, RESULTS
reflecting the final concentration in agglutination assays,
were expressed as inhibitor volume divided by total volume P1 antigenic activity of inhibitor preparations. Globoside
(for diluted whole egg white preparations) or as the molar did not inhibit agglutination of P1 erythrocytes by anti-P1
concentration (for ovomucoid and globoside). serum, demonstrating the absence of P1 antigenic activity. In
Incubation of erythrocytes with inhibitors. Human p eryth- contrast, the egg white preparations were potent inhibitors
rocytes were incubated for 1 h at room temperature or at of hemagglutination (MIC100s of 1:1.6 x 106 [CLEW and
37°C with PBS, the egg white preparations (undiluted CEW, PEW], 1:8 x 105 [DEW], and 50 pM [POM]), indicating
CLEW, DEW, and PEW or 500 ,uM POM), or globoside (2.5 high-level P1 antigenic activity. CEW exhibited weak hem-
,uM and serial twofold dilutions). The cells were then agglutination inhibition (MIC100, 1:6 x 103), indicating only
washed, resuspended in PBS with 10% (wt/vol) D-mannose, low-level P1 antigenic activity.
and used in agglutination assays with strain JJ48. Hemagglutinating activities of PEW and CEW. P1 erythro-
Neutralization of PEW agglutination inhibition by human cytes agglutinated in the presence of PEW and CEW inde-
anti-P1 serum. Serial dilutions of PEW and of POM were pendently of the presence of anti-P1 serum. This phenome-
prepared by using both commercial human anti-P1 and 5% non was not observed beyond a dilution of 1:3 (CEW) or 1:24
bovine serum albumin (BSA) in PBS and incubated over- (PEW) and appeared to be independent of the P blood group,
night at 4°C. These preparations were then tested as inhibi- since P1, P2, and p erythrocytes were agglutinated similarly.
tors of P bead agglutination by strain JJ48.
MR agglutination phenotypes of bacterial adhesins. In the
P1 antigenic activities of egg white preparations. The P1 presence of PBS, all control strains exhibited agglutination
patterns appropriate for their defined adhesin phenotypes
antigenic activities of the egg white preparations were quan- (Table 3). P-fimbrial strains JJ48 and HU849 demonstrated
tified by determining the MIC100s of these preparations for P-fimbrial adherence only (agglutination of P beads and of P1
agglutination of P1 erythrocytes by commercial anti-P1 se- but not p erythrocytes). The MAC of strain JJ48 for agglu-
rum. Equal volumes of inhibitor solution (prepared in serial tination of P beads or P1 erythrocytes was 60 to 120 times
twofold dilutions), anti-P1 serum, and a 2% (vol/vol) eryth- lower than the MAC of strain HU849, indicating a greater
rocyte suspension in PBS were incubated together at 4°C for P-fimbrial agglutinating capacity with the pUC derivative
15 min. After centrifugation at 1,000 x g for 15 s, cell pellets pJJ48 than with the pACYC184 derivative pRHU845. Strains
were gently resuspended and the intensity of agglutination with NPMR adhesins but no P fimbriae exhibited only
was noted. The MIC10o was defined as the lowest (final) NPMR adherence (agglutination of P1 and p erythrocytes but
concentration of inhibitor which still completely inhibited not of P beads), the hybrid strain with both P and NPMR
hemagglutination. fimbriae (JJ105) exhibited both P and NPMR adherenceVOL. 60, 1992 AVIAN P1 ANTIGENS INHIBIT P-FIMBRIAL AGGLUTINATION 581
TABLE 4. Comparative potencies of inhibitors of P-fimbrial in concentrations a low as 20 ,uM, whether incubated at
agglutination room temperature or at 37°C.
Dilution or concn (,iM) of following inhibitor":
Indicator" Endpoint DISCUSSION
CLEW DEW PEW POM Globoside
P1 RBC MIC50 1:64 1:54 1:45 30 10 We found that whole CLEW, DEW, and PEW, as well as
MICloo 1:19 1:14 1:14 80 50 purified POM, exhibited both potent P1 antigenic activities
and potent inhibitory activities against P-fimbrial agglutina-
P beads MIC50 1:45 1:45 1:43 20 20 tion. In contrast, CEW exhibited a 1,000-fold-lower level of
MICloo 1:14 1:14 1:12 90 130 P1 antigenic activity than the other whole egg whites and was
aP1 RBC, human P1 erythrocytes; P beads, digalactoside-coated latex a poor inhibitor of P-fimbrial adherence. This correlation of
beads. P1 antigenic activity with P-fimbrial inhibitory capacity,
b Values are final dilutions or final concentrations of inhibitors in aggluti- together with the partial neutralization of the P-fimbrial
nation assays (geometric mean of two to four determinations). POM and inhibitory activity of PEW and POM by anti-P1 serum,
globoside are purified inhibitors. Assays incorporated strain JJ48 at four times suggests that P1-antigen-containing egg whites inhibit P-fim-
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the MAC.
brial binding because of the P1 antigen present in their
ovomucoid fraction. These observations further suggest that
the P1 antigen present in avian ovomucoid is recognized not
(agglutination both of P beads and of P1 and p erythrocytes), only by anti-P1 serum but also by the receptor-binding site of
and the adhesin-negative strain (HB101) exhibited no agglu- the P-fimbrial adhesin molecule.
tination capacity at all (data not shown). The strain express- The MR adhesins of uropathogenic E. coli are commonly
ing pap-2-encoded fimbriae (P678-54/pJFK102) agglutinated considered in two categories, i.e., those with digalactoside
sheep erythrocytes strongly but human P1 erythrocytes binding specificity (P fimbriae), and the remainder, which
weakly and inconsistently, precluding use of this strain in recognize other receptors (Dr, AFA, S, G, and M adhesins
subsequent agglutination inhibition experiments with human and others of undetermined specificities) (12, 33). This
P1 erythrocytes. distinction is clinically significant because P fimbriae are
Agglutination inhibition experiments. Globoside com- associated with upper urinary tract colonization and pyelo-
pletely blocked agglutination of P beads and P1 erythrocytes nephritis (34), in contrast to NPMR adhesins, some of which
by the recombinant P-fimbriated strains but did not affect appear to play a more prominent role in lower urinary tract
agglutination of P1 or p erythrocytes by strains expressing infection (12). The laboratory detection of NPMR adhesins
NPMR adhesins (whether alone or together with P fimbriae) in the absence of P-fimbrial expression is straightforward
(Table 3). Of the egg white preparations, CEW failed to with the combined use of P bead agglutination and MR
inhibit agglutination completely with any adhesin. In con- hemagglutination assays: P beads are not agglutinated by
trast, CLEW, DEW, PEW, and POM fully inhibited all the NPMR adhesins, but human erythrocytes are. However,
P-fimbrial agglutination reactions inhibited by globoside. P-fimbriated strains agglutinate both P beads and P+ eryth-
Like globoside, these preparations had no effect on the rocytes, obscuring the possible presence of NPMR adhesins.
agglutination of P1 or p erythrocytes by strains expressing Use of an inhibitor of P-fimbrial hemagglutination such as
NPMR adhesins. None of the inhibitors blocked agglutina- globoside allows underlying NPMR-mediated hemagglutina-
tion of sheep erythrocytes by the strain expressing pap-2- tion to be observed with strains expressing both P and
encoded fimbriae (data not shown). NPMR adhesins. Unfortunately, globoside and similar diga-
Comparative agglutination inhibition capacities of inhibi- lactoside-containing glycolipids or oligosaccharides are pro-
tors. CLEW, DEW, and PEW were similarly potent inhibi- hibitively expensive ($100 to $500 for 5 mg is typical);
tors of P bead and P1 erythrocyte agglutination, giving furthermore, preliminary studies suggest that at least among
complete inhibition up to a final dilution of approximately wild-type E. coli strains from patients with urosepsis, glo-
1:14, and 50% inhibition at a dilution of approximately 1:50 boside is an unreliable inhibitor of P-fimbrial agglutination
(Table 4). POM and globoside also appeared approximately (12a). An alternative method for detecting NPMR adhesins
equipotent, giving complete inhibition up to a final concen- in the presence of P fimbriae by using MR agglutination of p
tration of from 50 to 130 ,uM and 50% inhibition at lt to 30 erythrocytes is impractical for general laboratory use be-
FLM (Table 4). cause of the restricted availability and the limited shelf life of
Neutralization of PEW and POM with human anti-P1 p erythrocytes. In this study, the P-fimbrial inhibitors
serum. After overnight incubation with human anti-P1 se- CLEW, DEW, PEW, and POM did not interfere with
rum, PEW and POM exhibited reduced inhibition of P bead hemagglutination mediated by NPMR adhesins, which per-
agglutination by strain JJ48, in comparison with control mitted identification of NPMR adherence in a strain express-
preparations of PEW and POM incubated overnight with 5% ing both P and NPMR adhesins (JJ105). This evidence
(wt/vol) BSA (data not shown). Neither of these PEW and suggests that CLEW, DEW, and PEW, as well as the
POM preparations agglutinated P beads in the absence of ovomucoid fractions thereof, could replace globoside as
bacteria. inhibitors of P-fimbrial binding and could obviate the need
Incubation of p erythrocytes with inhibitors. To determine for p erythrocytes in determining the presence or absence of
whether CLEW, DEW, PEW, or POM is incorporated onto NPMR adhesins in strains which also express P fimbriae.
the erythrocyte surface, p erythrocytes were incubated for 1 P1 antigenic activity has previously been identified in
h at room temperature or at 37°C with these egg white association with doves (6, 7), pigeons (2, 11, 30), and
substances, globoside, or PBS. Even after incubation at 37°C budgerigars (2, 6) but has been reported to be absent from
in egg white preparations above the MIC100 for inhibition of chickens, geese, turkeys, and ducks (2, 6). Our finding that
erythrocyte and P bead agglutination, p erythrocytes were CEW exhibited P1 antigenic activity, although at a much
not agglutinated by strain JJ48. In contrast, p erythrocytes lower level than CLEW, DEW, and PEW did, and the
were agglutinated by strain JJ48 after exposure to globoside finding that CEW decreased somewhat (but did not com-582 JOHNSON ET AL. INFECT. IMMUN.
pletely inhibit) P-fimbrial agglutination, suggest that previ- 13(Suppl.):S196-S200.
ously described differences between avian groups with re- 10. Hull, R. A., R. E. Gill, P. Hsu, B. H. Minshew, and S. Falkow.
spect to P1 antigen content may be a matter more of degree 1981. Construction and expression of recombinant plasmids
than of the absolute presence or absence of the antigen. encoding type 1 or D-mannose-resistant pili from a urinary tract
The abilities of globoside and other glycolipids to insert infection Escherichia coli isolate. Infect. Immun. 33:933-938.
10a.Hull, S. I. Personal communication.
into lipid membranes (17, 20), illustrated here by the finding 11. Inglis, G., A. G. Watt, and A. C. Munro. 1982. The production
that p erythrocytes were agglutinated by P-fimbriated bacte- of anti-P1 in rabbits immunised with pigeon ovomucoid. Med.
ria after but not before incubation with globoside, have been Lab. Sci. 39:179-181.
identified as a potential hazard in the use of glycolipids as 12. Johnson, J. R. 1991. Virulence factors in Escherichia coli
inhibitors of P-fimbrial binding in humans (9). More-severe urinary tract infection. Clin. Microbiol. Rev. 4:80-128.
urinary tract infections could result from the administration 12a.Johnson, J. R. Unpublished data.
of globoside if receptor density were to increase sufficiently 13. Johnson, J. R., P. L. Roberts, and W. E. Stamm. 1987. P-fim-
to offset the effect of adhesin blockade (9). We found that briae and other virulence factors in Escherichia coli urosepsis:
incubation with CLEW, DEW, PEW, or POM failed to association with patient's characteristics. J. Infect. Dis. 156:
225-229.
render p erythrocytes agglutinable by P-fimbriated bacteria, 14. Kallenius, G., R. Mollby, S. B. Svenson, J. Winberg, A. Lund-
Downloaded from http://iai.asm.org/ on February 6, 2021 by guest
probably because the Pl-antigen-containing ovomucoid mol- blad, S. Svensson, and B. Cedergren. 1980. The pk antigen as
ecule lacks a lipid moiety (corresponding with the ceramide receptor for the haemagglutinin of pyelonephritic Escherichia
portion of globoside) capable of inserting into the erythro- coli. FEMS Microbiol. Lett. 7:297-302.
cyte membrane (6, 7). These considerations suggest that 15. Kallenius, G., S. B. Svenson, H. Hultberg, R. Mollby, J. Win-
avian Pl-antigen-containing glycoproteins may be more suit- berg, and J. A. Roberts. 1983. P-fimbriae of pyelonephritogenic
able than digalactoside-containing glycolipids for studies of Escherichia coli: significance for reflux and renal scarring-a
pyelonephritis prevention in humans. hypothesis. Infection 1:73-76.
16. Kallenius, G., S. B. Svenson, R. Mollby, B. Cedergren, H.
Hultberg, and J. Winberg. 1981. Structure of carbohydrate part
ACKNOWLEDGMENTS of receptor on human uroepithelial cells for pyelonephritogenic
John Moulds provided information regarding the egg white-PI Escherichia coli. Lancet ii:604-606.
antigen connection, suggestions for the preparation of egg white 17. Kallenius, G., S. B. Svenson, R. Mollby, T. Korhonen, J.
solutions, and p erythrocytes. Sheila and Richard Hull (Department Winberg, B. Cedergren, I. Helin, and H. Hultberg. 1982. Carbo-
of Microbiology, Baylor College of Medicine, Houston, Tex.) pro- hydrate receptor structures recognized by uropathogenic E.
vided strains HU849 and HU968-115, Sima Bilge (Department of coli. Scand. J. Infect. Dis. 33:52-60.
Microbiology, University of Washington, Seattle, Wash.) provided 18. Karr, J. F., B. Nowicki, L. D. Truong, R. A. Hull, and S. I. Hull.
strain HB101/pSSS1, Bogdan Nowicki (Department of Obstetrics 1989. Purified P fimbriae from two cloned gene clusters of a
and Gynecology, University of Texas Medical Branch at Galveston, single pyelonephritogenic strain adhere to unique structures in
Galveston, Tex.) provided strain IH11128, and Harry L. T. Mobley the human kidney. Infect. Immun. 57:3594-3600.
provided strain 536-21. Denny Lyons, Melinda Walen, Bruce 19. Karr, J. F., B. J. Nowicki, L. D. Truong, R. A. Hull, J. J.
Redgate, and Steve Whiting provided pigeon, cockatiel, and dove Moulds, and S. I. Hull. 1990. pap-2-encoded fimbriae adhere to
eggs. Terriel Berggren constructed pJJ48, and both she and Allan the P blood group-related glycosphingolipid stage-specific em-
Ross assisted in the laboratory. Beth Wetak helped prepare the bryonic antigen 4 in the human kidney. Infect. Immun. 58:4055-
manuscript. 4062.
20. Leffler, H., and C. Svanborg Eden. 1980. Chemical identification
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