bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Escherichia coli and Bacteroides fragilis on peritoneal host defenses
Authors: Dunn D L , Barke R A , Ewald D C , Simmons R L
Published: January 2020

Escherichia coli and Bacteroides fragilis are common copathogens in clinical intra-abdominal sepsis, yet it is unclear how they interact synergistically in vivo. We sought to determine whether E. coli and B. fragilis, in combination but not alone, could exert a detrimental effect on the peritoneal host defenses of translymphatic absorption and bacterial phagocytosis. Our data indicated that nonviable E. coli (O18ab:K56/K7:- and O111:B4), Klebsiella pneumoniae, B. fragilis, and Bacteroides thetaiotaomicron were handled in a similar fashion by both host defenses of the peritoneal cavity. The use of 2 X 10(8) nonviable radiolabeled E. coli as a tracer and either 2 X 10(9) B. fragilis or 2 X 10(9) E. coli (either viable or nonviable) as a competing agent to inhibit host defenses demonstrated that although clearance and phagocytosis could be inhibited, the inhibition occurred to a similar degree with either E. coli or B. fragilis. Thus, B. fragilis did not compete to any greater extent than E. coli did for peritoneal clearance or opsonization and phagocytosis in vivo. These data indicate that bacterial synergy probably does not occur on the basis of reduced peritoneal clearance or by a reduction in the opsonization and phagocytosis of either organism by the copathogen. These results provide indirect support for the hypothesis that in bacterial synergy, one organism directly stimulates the growth of the other, perhaps by providing a growth factor.
Composition and antigenic activity of the oligosaccharide moiety of Haemophilus influenzae type b lipooligosaccharide
Authors: Inzana T J , Seifert W E , Williams R P
Published: January 2020

The oligosaccharide moiety of the lipooligosaccharide of Haemophilus influenzae type b strain Eag was isolated from the lipid component by mild acid hydrolysis and purified by gel filtration. Fast atom bombardment-mass spectrometry indicated that the lipid-free oligosaccharide had a basic molecular weight of 1,768; polysaccharides comparable to high-molecular-weight O side chains were not found. Glucose, galactose, galactosamine, heptose, 3-deoxy-D-manno-2-octulosonic acid (KDO), ethanolamine, and phosphate were identified in the lipid-free oligosaccharide by colorimetric assays, gas chromatography-mass spectrometry, or an amino acid analyzer. The presence of KDO was not clearly established by a thiobarbituric acid assay or by growth inhibition by a diazaborine derivative thought to block KDO synthesis. However, the semicarbizide assay and gas chromatography-mass spectrometry confirmed the presence of KDO. Lectin precipitation by Eag lipooligosaccharide in gels indicated that beta-D-galactose was present and that some of this monosaccharide was a terminal, nonreducing residue linked to N-acetyl-D-galactosamine. The lipid-free oligosaccharide was antigenic and completely inhibited lipooligosaccharide antibody (predominantly immunoglobulin G [IgG] and IgM) in an enzyme-linked immunosorbent assay, whereas the solubilized lipid A moiety did not. H. influenzae type b lipid-free oligosaccharide differed from core oligosaccharide of Salmonella lipooligosaccharide by the presence of galactosamine and a smaller percentage of heptose and KDO.
Role of Escherichia coli type 1 pilus in colonization of porcine ileum and its protective nature as a vaccine antigen in controlling colibacillosis
Authors: Jayappa H G , Goodnow R A , Geary S J
Published: January 2020

This study was designed to evaluate the role of Escherichia coli type 1 pili in adherence of the organism to porcine small intestines and the efficacy of pili as a vaccine antigen in controlling neonatal colibacillosis. Our results demonstrated that an E. coli phase cloned to express type 1 pili readily attached to the small intestines of colostrum-deprived newborn pigs. Immunofluorescent staining of intestine sections revealed the presence of E. coli expressing type 1 pili only on the brush border, suggesting involvement of type 1 pili in the colonization process. Administration of anti-type 1 serum to newborn pigs prior to challenge reduced the level of gut-associated E. coli sixfold compared with controls. Purified type 1 pilus vaccine induced significant protection against colibacillosis in newborn pigs following challenge with E. coli expressing type 1 pili. Pigs born to vaccinated gilts scoured less and gained more weight than pigs born to control gilts. Our results demonstrate that type 1 pili are a virulence factor, as well as an effective vaccine antigen.
Adaptation of Ehrlichia sennetsu to canine blood monocytes: preliminary structural and serological studies with cell culture-derived Ehrlichia sennetsu
Authors: Holland C J , Ristic M , Huxsoll D L , Cole A I , Rapmund G
Published: January 2020

Ehrlichia sennetsu, the causative agent of human sennetsu rickettsiosis, was successfully propagated in primary canine blood monocyte cultures. The growth cycle of this organism appears to be similar to that of Ehrlichia canis. The antigen derived from our E. sennetsu cultures was used to develop an indirect fluorescent antibody test for detection and titration of serum antibodies to the organism. Using this test system, we found that five human serum samples obtained from patients clinically diagnosed as having sennetsu rickettsiosis were positive for anti-E. sennetsu antibodies. In addition, 29% of the serum samples obtained from 200 patients having a fever of unknown origin and residing in various regions of Malaysia were also serologically positive. All sera from apparently healthy individuals were negative in the test. Dogs inoculated with cell culture-adapted E. sennetsu developed a significant specific antibody titer to E. sennetsu, and the organism was subsequently isolated from their blood. These animals showed no clinical evidence of disease. The possibility of a higher prevalence of human sennetsu rickettsiosis in Southeast Asia and the potential usefulness of the canine model for studies of human sennetsu rickettsiosis are discussed.
Nonimmune binding of equine immunoglobulin by the causative organism of contagious equine metritis, Taylorella equigenitalis
Authors: Widders P R , Stokes C R , Newby T J , Bourne F J
Published: January 2020

This study identifies nonimmune binding of equine immunoglobulin by the causative organism of contagious equine metritis. Immunoglobulin binding to the bacterium was strongest for immunoglobulin G (IgG) and less for IgM; IgA was not bound. Binding of equine IgG was inhibited by human IgG, but not by IgG of domestic animals. Immunoglobulin binding by the bacterium appeared to be directed towards an epitope in the hinge region of the immunoglobulin molecule.
Serotypes, hemolysin production, and receptor recognition of Escherichia coli strains associated with neonatal sepsis and meningitis
Authors: Korhonen T K , Valtonen M V , Parkkinen J , Väisänen-Rhen V , Finne J , Orskov F , Orskov I , Svenson S B , Mäkelä P H
Published: January 2020

Sixty-three Escherichia coli strains isolated from neonatal sepsis or meningitis were studied and compared with previous data on fecal or urinary pyelonephritis-associated isolates from children. Characteristics significantly associated with neonatal infection were capsular type K1 (54%), O group 18 (27%), rough-type lipopolysaccharide together with K1 capsule (19%), and S fimbriae (29%). Within the neonatal infection group, the K1 capsule and rough lipopolysaccharide were most common among the youngest infants (0 to 21 days old) and in meningitis. Hemolysin production, P fimbriae, and X adhesions (adhesions not identifiable as type 1, P, or S) were significantly more common in the two materials from infections as compared with the fecal isolates. One large clone of 11 strains (O18:K1:H7, with both type 1 and S fimbriae) and three smaller ones (O7:K1:H1 and O6:K2:H1, both with type 1 and P fimbriae and X adhesions; and R:K1:H33 with no adhesions) were identified among the strains from neonatal infections. Only O6:K2:H1 strains were also common among the strains from pyelonephritis.
Antigenic heterogeneity of lipopolysaccharides from Campylobacter jejuni and Campylobacter fetus
Authors: Perez G I , Hopkins J A , Blaser M J
Published: January 2020

The lipopolysaccharide (LPS) structure of Campylobacter spp. can be visualized with polyacrylamide gel electrophoresis by examining proteinase K-treated whole cell lysates. Polyacrylamide gel electrophoresis LPS profiles of C. jejuni strains are rough type with low concentrations of low-molecular-weight polysaccharide side chains, serum-resistant C. fetus strains have smooth-type LPS, and serum-sensitive C. fetus strains have rough-type LPS. We electroblotted the proteinase K-treated whole cell lysates of 17 C. jejuni and 9 C. fetus strains from polyacrylamide gel electrophoresis to nitrocellulose paper to examine antigenicity to immune rabbit sera. There was virtually no antigenic cross-reactivity of C. jejuni and C. fetus LPS. Among C. jejuni strains, core LPS structures were cross-reactive, but the O-polysaccharide side chains were best recognized by homologous antisera. Antisera to several serum-resistant C. fetus strains recognized only the polysaccharide side-chain regions of serum-resistant strains and no part of the LPS from the sensitive strain. Antiserum raised against a serum-sensitive C. fetus strain but not homologous antisera recognized the core region of the LPS of the serum-resistant C. fetus strains. These findings suggest that core LPS antigens are widely shared within C. fetus subsp. fetus strains but that in the serum-resistant strains this core region is not surface exposed and therefore not immunogenic to rabbits infected with whole cells.
Effect of subgingival scaling on systemic antibody responses to oral microorganisms
Authors: Ebersole J L , Taubman M A , Smith D J , Haffajee A D
Published: January 2020

The effects of scaling and root planing treatment on systemic antibody responses were studied in patients with periodontal disease and in normal subjects. Immunoglobulin G antibody in serum to a battery of oral microorganisms was assessed in an enzyme-linked immunosorbent assay before and after treatment in 31 individuals. The majority (96%) of the diseased patients exhibited elevated antibody to one or more of the microorganisms before the scaling regime. Significant increases in antibody levels in serum were noted in 16 of 19 patients after scaling, whereas only 2 of 12 nonscaled subjects showed similar changes during monitoring intervals of up to 3 years. The bacterial specificities of the increases were found to differ among the patients; however, a significant correlation to preexisting elevated antibody levels was observed. Peak levels of responses were noted at approximately 2 to 4 months posttreatment; antibody returned to pretreatment levels by 8 to 12 months. The predominant organisms for which changes were noted included the black-pigmented Bacteroides spp., Eikenella corrodens, Campylobacter concisus, and Actinobacillus actinomycetemcomitans. In 18 of 19 instances, the homologous microorganism was detected in the subgingival plaque when elevated antibody was present after treatment. These findings indicated that specific changes in host systemic responses accompany scaling and root planing treatment of periodontal disease patients. These alterations in the host response may provide an additional means by which successful therapy can be accomplished.
Enhanced nasopharyngeal colonization of rats by piliated Haemophilus influenzae type b
Authors: Anderson P W , Pichichero M E , Connor E M
Published: January 2020

Piliated Haemophilus influenzae type b strains display an enhanced adherence to human epithelial cells in vitro. However, clinical isolates, even from mucosal sites, are seldom piliated, although piliated populations can be selected from them. Experiments with rats have led some authors to suggest that piliation does not implement colonization by H. influenzae type b. Piliated populations were obtained from 35 strains by selection for adherence to human erythrocytes. One strain, H. influenzae H305, simultaneously acquired an increased adherence to rat erythrocytes and buccal epithelial cells. In contrast to other strains, H. influenzae H305 in piliated form was more effective than in nonpiliated form in the colonization of rats by intranasal inoculation. After the piliated inoculum, however, the colonies cultured from the nasal washes were negative for erythrocyte adherence. Thus, piliated H. influenzae type b strains have an apparent advantage to initiating colonization in the rat model but may give rise to nonpiliated progeny that are more readily cultivable from the mucosal surface.
Effect of calcium chloride on experimental infection of mice with Pseudomonas aeruginosa
Authors: Tamura Y , Tanaka S
Published: January 2020

Concomitant administration of Pseudomonas aeruginosa and calcium chloride to mice enhanced the virulence of some strains. The 50% lethal dose of P. aeruginosa 5 decreased by more than three orders of magnitude, regardless of the challenge route (intramuscular, subcutaneous, and intraperitoneal injection), while the 50% lethal dose of strain N10 did not decrease. When challenged intramuscularly with 10(4) organisms of strain 5 or strain N10 mixed with calcium chloride, both strains multiplied at the local site of injection. Strain 5 subsequently caused systemic infection, while strain N10 did not. The virulence-enhancing effect of calcium chloride can be successfully applied in protection assays for immunized mice challenged with strain 5.
Ribosomal protein phosphorylation induced during Q fever or by lipopolysaccharide: in vitro translation is stimulated by infected liver ribosomes
Authors: Hickey M J , Gonzales F R , Paretsky D
Published: January 2020

Q fever, as well as the lipopolysaccharide prepared from the rickettsial agent Coxiella burnetii, stimulates the phosphorylation of guinea pig liver ribosomal protein S6. In vitro mRNA and ribosome-dependent rabbit reticulocyte lysate translation systems reconstituted with ribosomes and mRNAs from infected animal livers were more active than those with mRNAs and ribosomes from uninfected animals. Treatment of ribosomes with a ribosomal supernatant phosphatase reduced the in vitro translation activities; the largest decreases occurred in systems with ribosomes and mRNAs from infected liver. These experiments provide a basis for explaining the increased hepatic protein synthesis during Q fever and demonstrate, perhaps for the first time, the phosphorylation of ribosomal protein in response to lipopolysaccharide. The implications of these observations are discussed in the context of previous studies on stimulated transcription and translation during Q fever.
Complement activation and attack on autologous cell membranes induced by streptolysin-O
Authors: Bhakdi S , Tranum-Jensen J
Published: January 2020

Streptolysin-O damages mammalian membranes through generation of large transmembrane channels formed by membrane-inserted polymers of the toxin (S. Bhakdi et al., Infect. Immun. 47:52-60, 1985). We here report that the native toxin binds naturally occurring human serum immunoglobulin G antibodies to form immune complexes with potent complement-activating capacity. Nanomolar concentrations of toxin added to antibody-containing serum cause rapid consumption of C4 and C5 hemolytic activity and 30 to 90% C3 conversion within 10 to 60 min at 37 degrees C. After binding to target membranes, streptolysin-O polymers serve as foci for antibody-dependent complement activation, which proceeds to completion with the formation of terminal C5b-9 complexes on the autologous cells. The binding and insertion of a primarily water-soluble bacterial product into a host cell membrane has thus been shown to generate a stable and hyperactive focus for activation of and self-attack by the complement system. We suggest that this process perpetuates local tissue damage, deviates host complement action away from the invading bacteria, and may possibly play a role in the pathogenesis of poststreptococcal disease.
Evaluation of BW942C, a novel antidiarrheal agent, against enterotoxins of Escherichia coli and Vibrio cholerae
Authors: Morgan D R , Sellin J , Gutierrez L , DuPont H L , Wood L V
Published: January 2020

BW942C, an enkephalin-like pentapeptide with anti-diarrheal activity, was tested against crude toxins of Escherichia coli and Vibrio cholerae in the Y-1 adrenal cell assay, rabbit ileal loop assay, and suckling mouse assay. The effects of BW942C on in vitro ion transport were measured in rabbit ileum mounted in Ussing chambers. In vitro, BW942C decreased basal short-circuit current (2.26 and 3.15 mueq cm-2 h-1 in experimental samples and controls, respectively; n = 7, P less than 0.05) and increased basal net Cl absorption (1.59 and 0.50 mueq cm-2 h-1 in experimental samples and controls, respectively; P less than 0.025). Net Na absorption was also increased, but not significantly. BW942C did not block the secretory response to a maximal dose of purified heat-stable toxin. BW942C directly enhanced intestinal fluid absorption. In the Y-1 adrenal cell assay, 5 mg of BW942C per ml inhibited the cytopathic effect caused by cholera toxin or heat-labile enterotoxin of E. coli. In the rabbit ileal loop assay, E. coli heat-stable toxin, E. coli heat-labile enterotoxin, and cholera toxin were inhibited 35 to 70% by administration of BW942C. With the suckling mouse model, the fluid accumulation caused by E. coli heat-stable toxin was ablated by prior treatment with BW942C. The drug is currently being evaluated in patients with acute secretory diarrhea to determine its effect on clinical symptoms.
Cross-reacting human and rabbit antibodies to antigens of Histoplasma capsulatum, Candida albicans, and Saccharomyces cerevisiae
Authors: Kumar B V , Medoff G , Kobayashi G S , Sieling W L
Published: January 2020

Using Western blots of electrophoretically separated antigens, we show that human antibodies react most frequently to antigens shared by three fungi (Histoplasma capsulatum, Candida albicans, and Saccharomyces cerevisiae). Reactivity to antigens specific for individual fungi was relatively uncommon. The pattern of reactivity could not distinguish infected patients from uninfected controls. Rabbits immunized with extracts of each fungus also produced antibodies to cross-reactive or shared antigens of the other two fungi. Furthermore, preimmune sera showed similar but lower reactivity with the same fungal antigens. We believe that the preimmunization antibodies, which probably resulted from earlier fungal colonization or inapparent infections, predisposed the immune responses elicited by the vaccinations. A similar mechanism likely explains the results with human sera.
Adhesion of enterotoxigenic Escherichia coli to human small intestinal enterocytes
Authors: Knutton S , Lloyd D R , Candy D C , McNeish A S
Published: January 2020

An improved enterocyte adhesion assay has been used to examine a collection of 44 strains of enterotoxigenic Escherichia coli (ETEC) for their ability to adhere to the brush border of isolated human duodenal enterocytes. Fourteen strains showed good adhesion; in each case the ability to adhere correlated with the production of colonization factor antigen I or II (CFA/I or CFA/II) fimbriae. CFA/II-positive producing coli surface antigens 1 and 3 (CS1 and CS3), coli surface antigens 2 and 3 (CS2 and CS3), and only coli surface antigen 3 (CS3) each showed good adhesion. CS3-mediated brush border attachment of CFA/II-positive ETEC was demonstrated by electron microscopy with monospecific antibody and an immunogold labeling technique. One CFA/I-positive ETEC strain was nonadherent in the assay, as were ETEC producing type 1 somatic fimbriae. Five animal ETEC strains producing K88, K99, F41, and 987P fimbriae were slightly more adhesive than control strains, but adhesion was significantly less than that of CFA-positive ETEC. Twenty five human ETEC strains that lacked CFA/I and CFA/II were nonadherent, suggesting either that the surface antigens responsible for adhesion to human intestinal mucosa in these strains were not being produced or that mucosal receptors for these strains are present in regions of the small intestine other than the duodenum.
Suppression of lymphocyte proliferation by Pseudomonas aeruginosa: mediation by Pseudomonas-activated suppressor monocytes
Authors: Issekutz T B , Stoltz J M
Published: January 2020

Pseudomonas aeruginosa has been shown to suppress cell-mediated immunity in experimental animals, but recent reports have also demonstrated that there is a strong T-cell response to this bacteria. Our studies of human peripheral blood mononuclear cells showed a great variation in the in vitro proliferative response to killed P. aeruginosa, so we examined the interaction of the different mononuclear cells in cultures with this bacteria. P. aeruginosa stimulated the proliferation of T lymphocytes, specifically the surface-immunoglobulin-negative, T8- subset, which are felt to be T helper cells. P. aeruginosa added in coculture experiments to peripheral blood mononuclear cells stimulated with Staphylococcus aureus, Streptococcus pyogenes, or tetanus toxoid suppressed the proliferation to these latter antigens. This proliferation was not affected by the depletion of adherent monocytes from peripheral blood mononuclear cells, and the suppression was restored when monocytes were added back to these cultures. Moreover, monocytes pulsed with P. aeruginosa but not with S. aureus suppressed the antigen-induced proliferation of peripheral blood mononuclear cells. This monocyte suppression was not inhibited by indomethacin and was unlikely to be the result of prostaglandin synthesis by these cells. Thus, P. aeruginosa can induce monocytes to suppress antigen-stimulated T-lymphocyte proliferation in vitro, and these suppressor cells may facilitate the growth of this organism in disorders such as cystic fibrosis.
Antibody-independent and -dependent opsonization of group B Streptococcus requires the first component of complement C1
Authors: Levy N J , Kasper D L
Published: January 2020

The role of the classical complement pathway and specifically the first component, C1 in antibody-independent opsonization of type Ia group B Streptococcus (GBS) was investigated. For these studies a radiolabeled bacterial uptake assay was developed that was dependent on time and bacterial concentration and that required an intact classical complement pathway. To directly investigate the role of C1 in opsonization of type Ia GBS, C1 was isolated by chromatography on an immunoglobulin G (IgG) affinity column and further purified by molecular sieve chromatography on an Ultrogel AcA 22 column. When normal human serum was absorbed with 10(9) CFU of type Ia or III GBS, the serum opsonic capacity diminished (33 to 34%) for type Ia GBS compared with unadsorbed serum. Preincubation of the bacteria with purified C1 (10(4)U of C1 per ml) restored the opsonizing capacity of the adsorbed serum. A C1-depleted serum was prepared from the nonadherent fractions of the CH-sepharose 4B IgG column which only contained 5 U of C1 per ml. Substitution of C1-depleted reagent for normal serum in the uptake assay resulted in dramatic decreases in the opsonization of type Ia GBS, but opsonization could be restored by preincubation of the bacteria with purified C1. Heat-inactivated C1 depleted serum did not support opsonization of type Ia GBS, even with the addition of C1. Preincubation of type Ia GBS with heat-inactivated hyperimmune sera did not result in opsonization of type Ia GBS in the presence of C1-depleted serum. However, opsonization could be restored by the addition of C1, and the effects of C1 and antibody were additive. These results indicate the critical role of C1 in direct activation of the classical complement pathway by type Ia GBS and in antibody-mediated opsonization of the bacteria.
Role of resident macrophages, peripheral neutrophils, and translymphatic absorption in bacterial clearance from the peritoneal cavity
Authors: Dunn D L , Barke R A , Knight N B , Humphrey E W , Simmons R L
Published: January 2020

Microbial pathogens within the peritoneal cavity are thought to encounter three categories of host defense mechanisms: (i) removal mechanisms, which occur via diaphragmatic lymphatic absorption; (ii) killing mechanisms, in which host phagocytes act as effector cells; and (iii) sequestration mechanisms due to fibrin trapping and the formation of adhesions between visceral surfaces. We sought to define and quantitate the relative role of the first two components in an experimental rat model of Escherichia coli peritonitis in which fibrinous adhesions do not form. Intraperitoneal challenge with greater than or equal to 2 X 10(8) CFU of viable E. coli led to an initial decline in bacterial numbers followed by ongoing proliferation and greater than 50% mortality. With inocula of less than or equal to 5 X 10(7) CFU, elimination of bacteria occurred after moderate initial proliferation, and no mortality ensued. Nonviable, radiolabeled E. coli organisms were utilized to examine bacterial clearance via translymphatic absorption and phagocytosis. Both processes were extremely rapid, serving to eliminate free bacteria rapidly within the peritoneal cavity. Although macrophages and polymorphonuclear leukocytes within the peritoneal cavity demonstrated similar phagocytic capacities, the predominance of macrophages at the time of the initial bacterial insult led to the conclusion that these cells, in addition to translymphatic absorption, represent the first line of host defenses, acting to eliminate bacteria in the incipient stages of infection.
Enhanced oxidative burst in immunologically activated but not elicited polymorphonuclear leukocytes correlates with fungicidal activity
Authors: Brummer E , Sugar A M , Stevens D A
Published: January 2020

Polymorphonuclear neutrophils (PMN) induced locally in immune mice by intraperitoneal injection of antigen exhibit enhanced fungicidal activity compared with PMN elicited with thioglycolate. The mechanism of the differences in these PMN populations was studied. Sublethal infection was used to produce immunity to Blastomyces dermatitidis. A correlation was sought between the ability of PMN to kill, or not kill, B. dermatitidis and the production of the oxidative burst, as measured by luminol-enhanced chemiluminescence (CL). Although elicited PMN cocultured with Candida albicans produced a burst of CL and were candidacidal, killing did not occur when PMN were cocultured with B. dermatitidis. Lack of killing of B. dermatitidis by elicited PMN correlated with lack of stimulation of a brisk oxidative burst. In contrast to elicited PMN, PMN induced by B. dermatitidis antigen responded to this fungus with a burst of CL and a significant reduction of inoculum CFU (80%). Furthermore, these PMN when cocultured with C. albicans produced an enhanced burst of CL, and killing was enhanced compared with that by elicited PMN, e.g., 86 versus 58%. The CL burst and killing of B. dermatitidis by antigen-induced PMN was abrogated in the presence of catalase, implying a critical role for hydrogen peroxide. Partial but significant depression of CL and killing in the presence of dimethyl sulfoxide, a hydroxyl radical scavenger, identified hydroxyl radical, or its metabolites, as a toxic product(s) responsible for a significant fraction of fungicidal activity. These results indicate that the metabolic activity and microbicidal activity of PMN can be altered (enhanced) at the site of an immunological reaction and thus could constitute an important factor in resistance.
Genetic diversity in relation to serotype in Escherichia coli
Authors: Caugant D A , Levin B R , Orskov I , Orskov F , Svanborg Eden C , Selander R K
Published: January 2020

The extent of chromosomal-gene diversity among 261 isolates of Escherichia coli sharing single O, K, or H antigens and various combinations thereof was estimated by multilocus enzyme electrophoresis, which detects allelic variation in structural genes. The results of this study indicate that the genetic diversity among isolates sharing single antigenic determinants can approach or equal that observed among randomly chosen strains; that the magnitude of the diversity varies among antigens; and that the genetic diversity is reduced, but not eliminated, among isolates sharing two antigenic determinants. With one exception, isolates of the same O:K:H serotype were of identical or closely related electrophoretic types (ETs). Isolates of the same ET generally shared the same combination of antigenic determinants, but some ETs included isolates of different serotypes. The implications of these findings for epidemiological research and the clone hypothesis of population structure are discussed, and possible evolutionary mechanisms causing antigenic divergence and convergence are considered.
Isotype commitment of B cells and dissemination of the primed state after mucosal stimulation with Mycoplasma pulmonis
Authors: Rose F V , Cebra J J
Published: January 2020

Live Mycoplasma pulmonis organisms were used to examine the immune response in the bronchus-associated lymphoid tissue after primary and secondary challenge with M. pulmonis, to study the dissemination of the primed state to distal tissues (i.e., spleen, peripheral blood, and Peyer's patches), and to determine whether the chronic antigenic stimulation accompanying infection influences the isotype potential and commitment of the primed B cells recovered from the various tissues. We have shown that exposure to M. pulmonis by a variety of routes results in a generalized rise in frequency of T-dependent, antigen-sensitive B cells in all lymphoid tissues. The route of secondary exposure to M. pulmonis was found to markedly increase the frequency of M. pulmonis-specific B cells in the bronchus-associated lymphoid tissue relative to that in the Peyer's patches after intraduodenal but not intratracheal challenge. A substantial rise in the number of M. pulmonis-sensitive B cells in the peripheral blood suggests that the dissemination of the primed state, at least in part, is due to B-cell migration via lymph and blood from local sites exposed to M. pulmonis. The majority of T-dependent clones generated by M. pulmonis-specific B cells secrete exclusively immunoglobulin G1 (IgG1). We have demonstrated that the exaggerated IgG1 response was not due to the accompanying viable donor T cells in the inoculum. The predominance of IgG1 was also demonstrated in clones from the bronchus-associated lymphoid tissue of athymic BALB/c mice that were primed with M. pulmonis. Thus, we can infer that functional T cells are not required for the development of specific B cells with the potential for IgG1 expression at the time of in vivo priming. When anti-trinitrophenyl- and anti-M. pulmonis-specific clones were generated in the same splenic fragment cultures stimulated by trinitrophenylated M. pulmonis, only the M. pulmonis-specific clones showed exaggerated IgG1 expression. Therefore, we conclude that the exaggerated IgG1 response accompanying M. pulmonis infection of euthymic mice seems to be dependent, at least in part, on an intrinsic property of the B cells that develop during this antigenic stimulation.
Enhancement of enterotoxin production by carbon dioxide in Vibrio cholerae
Authors: Shimamura T , Watanabe S , Sasaki S
Published: January 2020

We found that Vibrio cholerae 569B produced much more cholera enterotoxin in the presence of added carbon dioxide than in its absence. An atmosphere of 10% carbon dioxide was optimal for maximal enterotoxin production.
Detection of Mycoplasma pneumoniae adhesin (P1) in the nonhemadsorbing population of virulent Mycoplasma pneumoniae
Authors: Kahane I , Tucker S , Baseman J B
Published: January 2020

Mycoplasma pneumoniae organisms possessing a hemadsorbing-negative (HA-) phenotype comprise more than 50% of the population of virulent M. pneumoniae cultures. Monoclonal antibody to P1, the major adhesin of M. pneumoniae reacts with this HA- mycoplasma fraction based upon radioimmunoprecipitation and immunoblotting. Demonstration of P1 in the entire mycoplasma population suggests that topological organization of this adhesin in the membrane or the physiological state of the mycoplasmas may determine hemadsorbing capabilities.
Heterogeneity of Trichomonas vaginalis and discrimination among trichomonal isolates and subpopulations with sera of patients and experimentally infected mice
Authors: Alderete J F , Suprun-Brown L , Kasmala L , Smith J , Spence M
Published: January 2020

The antibody response in trichomoniasis patients was examined with a variety of methodologies including enzyme-linked immunosorbent assays, indirect immunofluorescence, immunoblotting, and radioimmunoprecipitation-electrophoresis-autoradiography. Based on enzyme-linked immunosorbent assay recognition of trichomonal isolates, sera from patients with trichomoniasis were categorized into reactive class I (IA, IB, and IC) and nonreactive class II sera. A diminished ability to precipitate antibody-binding trichomonad membrane proteins by the whole cell radioimmunoprecipitation assay was noted from class IA to class II sera. The antigenic distinctions among various Trichomonas vaginalis isolates appeared due to high-molecular-weight protein antigens detected by class IA sera in a whole cell radioimmunoprecipitation assay. The heterogeneity in antigenic patterns was confirmed among the isolates with sera from experimental animals. Also, live T. vaginalis cells appear to have only a few of the entire repertoire of major immunogenic surface proteins accessible to antibody binding. Immunoblotting demonstrated that the high-molecular-weight proteins responsible for trichomonal isolate heterogeneity are present in all isolates. The data suggest that trichomonads of a given isolate express only a subset of internally synthesized protein antigens on their surface. Importantly, the presence of these protein antigens on T. vaginalis membranes correlated with antibody production in subcutaneously challenged mice. Finally, indirect immunofluorescence studies with highly reactive, pooled sera from either patients or mice revealed a subpopulation of nonstaining trichomonads. These data support the view that heterogeneity among T. vaginalis is dependent upon the surface disposition of highly immunogenic protein antigens. Strategies may now be developed not only for studying potential vaccine reagents, but also for examining possible antigenic phenotypic variations in this experimental model.
Relationships between levels of lysozyme, lactoferrin, salivary peroxidase, and secretory immunoglobulin A in stimulated parotid saliva
Authors: Rudney J D , Smith Q T
Published: January 2020

Recent studies suggest that salivary lysozyme (Lz), lactoferrin (Lf), peroxidase (Spx), and secretory immunoglobulin A (sIgA) may interact in a common antimicrobial system. A multiple protein approach therefore may be needed to determine the role of this system in oral health and ecology. In the present study we investigate the relationships between levels of Lz, Lf, Spx, and sIgA (adjusted for flow rate and total protein) in stimulated parotid saliva from 44 dental students. Principal components analysis was used to determine major patterns of intercorrelation between variables; cluster analysis was used to identify groups of subjects with similar salivary profiles for Lz, Lf, Spx, and sIgA. Spx tended to vary independently of Lz and Lf, which, in turn, tended to vary together. sIgA showed a weak negative relationship with Spx and a weak positive relationship with Lz and Lf. Six major clusters of subjects with similar antimicrobial protein profiles were found. These were significantly different at P less than 0.0001. Spx was the most important determinant of cluster membership followed (in order of importance) by Lz, Lf, and sIgA. Cluster profiles were Spx-, sIgAmu, Lf-, Lz-; Spx-, sIgA+, Lfmu, Lz+; Spxmu, sIgAmu, Lfmu, Lzmu; Spx+, sIgA-, Lf-, Lz-; Spx+, sIgAmu, Lf+, Lz-; and Spx+, sIgAmu, Lf+, Lz+ (-, mu, and + refer to the position of the cluster mean each variable relative to the overall mean for that variable). Results suggest that clusters may be a product of independent variation in the secretory activity of acinar and intercalated duct cells.
Relationship between gingival crevicular fluid and serum antibody titers in young adults with generalized and localized periodontitis
Authors: Tew J G , Marshall D R , Burmeister J A , Ranney R R
Published: January 2020

The objective of the present study was to determine the relationship between concentrations of antibodies in serum and those in gingival crevicular fluid (GCF) of patients with juvenile periodontitis and severe periodontitis. Most antigens used to quantitate antibodies were obtained from a panel of bacteria associated with juvenile periodontitis or severe periodontitis. We further investigated variation in antibody titer among different periodontal sites and the extent to which antibody in GCF is locally derived. Titers of antibody, total immunoglobulin G (IgG), and human serum albumin were determined with sensitive radioimmunoassays. The relationship between serum and GCF antibody was complex. Both person-to-person variability and marked variability within the same subject were found among different sites of similar clinical status. The site-to-site variability was found not only for antibody reactive with periodontal organisms, but also for antitetanus toxoid, total IgG, and even human serum albumin. Generally the variability was in the degree of depression of the level in GCF relative to that in serum. However, anti-Bacteroides gingivalis and anti-Actinobacillus actinomycetemcomitans in GCF often exceeded the level in serum. When antibody titers in serum and GCF were calculated per milligram of human serum albumin, most of the apparent depressions of antibody in GCF disappeared. The ratio of antibody in serum to that in GCF approached unity for all organisms except B. gingivalis and A. actinomycetemcomitans Y4, which were markedly elevated. Furthermore, the level of IgG per milligram of human serum albumin in GCF was about twice the level in serum. We believe that human serum albumin reflects serum contribution to the GCF, and we therefore attribute the increased level of IgG per milligram of albumin in GCF to local synthesis. It appears that anti-B. gingivalis and anti-A. actinomycetemcomitans represent an important portion of this local antibody synthesis, since most seropositive patients with severe or juvenile periodontitis had at least one site elevated, and the magnitudes of the elevations were large in many sites. Those sites yielding elevated antibody exhibited no obvious differences in clinical parameters of probeable depth or attachment level as compared with sites in which antibody levels in GCF were similar to serum levels. Elevated antibody in GCF may relate to changes in disease activity that are not detectable by usual clinical measures.
Causative ehrlichial organisms in Potomac horse fever
Authors: Rikihisa Y , Perry B D
Published: January 2020

An ehrlichia was consistently isolated from the peripheral blood leukocyte fraction of ponies that had been experimentally infected with Potomac horse fever by whole blood transfusion from naturally infected horses. The organism was propagated in a human histiocyte cell line for 3 to 5 weeks and then inoculated intravenously or intradermally into healthy adult ponies. Clinical signs of Potomac horse fever, which varied in the degree of severity, occurred 9 to 14 days post-inoculation in all of the ponies. One pony died 20 days post-inoculation. The ehrlichial organism was reisolated in the human histiocyte cell line from the blood leukocyte fraction of all of the experimental ponies on each day that samples were examined (days 9, 10, 11, 19, and 39). These organisms were identical to those originally detected in the wall of the intestine of ponies with clinically diagnosed Potomac horse fever when compared by light and electron microscopy and an immunofluorescence labeling technique. The immunofluorescent antibody titer became positive in a pony at 20 days postinjection. These results indicate that the ehrlichial organisms is the causative agent of Potomac horse fever.
Host response to infection with a temperature-sensitive mutant of Salmonella typhimurium in a susceptible and a resistant strain of mice
Authors: Nauciel C , Vilde F , Ronco E
Published: January 2020

The inoculation of a temperature-sensitive mutant of Salmonella typhimurium induced a long-lasting infection in susceptible (C57BL/6) and resistant (A/J) mice. During week 1 of infection, the number of bacteria in the spleens was similar in both mouse strains. Then, the decrease of bacteria was more rapid in the resistant strain. Splenomegaly and granulomatous hepatitis were more severe in the susceptible strain. The immune response induced by this infection was studied. In both mouse strains delayed-type hypersensitivity to Salmonella antigens was present, and resistance to reinfection with a virulent strain of S. typhimurium or with Listeria monocytogenes appeared with the same kinetics. Thus, it does not seem that the gene(s) controlling natural resistance to S. typhimurium act(s) on acquired immunity.
Purification and partial characterization of outer membrane proteins P5 and P6 from Haemophilus influenzae type b
Authors: Munson R S , Granoff D M
Published: January 2020

The major outer membrane proteins of Haemophilus influenzae type b (Hib), designated P5 and P6 (R.S. Munson, Jr., J.L. Shenep, S.J. Barenkamp, and D.M. Granoff, J. Clin. Invest. 72:677-684, 1983), were purified to homogeneity and partially characterized. P5 was insoluble in octylglucoside-NaCl and could be extracted with 1% sodium dodecyl sulfate (SDS) in 20 mM phosphate (pH 7.5). Solubilized P5 was further purified on hydroxylapatite in 0.1% SDS. The purified protein had an apparent molecular weight of 27,000 as determined by SDS-polyacrylamide gel electrophoresis after sample preparation at room temperature. The protein migrated with an apparent molecular weight of 35,000 after heating for 30 min at 100 degrees C in the presence of 10% beta-mercaptoethanol (beta ME). Rabbit antisera prepared against the purified preparation immunoprecipitated solubilized protein P5 but had no protective activity in the infant rat bacteremic model. The SDS-insoluble residue was further extracted with 1% SDS-0.5 M NaCl-0.1% beta ME at 37 degrees C. A single outer membrane protein, designated P6, with an apparent molecular weight of 16,000, remained insoluble under these conditions. Antiserum prepared against this insoluble fraction contained antibodies which, after removal of anti-lipopolysaccharide antibody, immunoprecipitated P6 and protected infant rats challenged with Hib. Protein P6 could be released from the insoluble cell wall in the presence of SDS-NaCl-beta ME at 60 degrees C. Thus, proteins P5 and P6 could be purified from the cell envelope of Hib. Based on the results from infant rat passive protection experiments, antigens in the P6-cell wall fraction merit further investigation as possible vaccine components. In contrast, epitopes on protein P5 did not appear to elicit protective antibody.
Identification of chitin as a structural component of Giardia cysts
Authors: Ward H D , Alroy J , Lev B I , Keusch G T , Pereira M E
Published: January 2020

The intestinal parasite Giardia lamblia is a significant cause of diarrheal disease, which is perpetuated by the infective cyst form of the parasite. Although a rational approach to the control of giardiasis would be to inhibit cyst formation, nothing is known of the chemical composition of the cyst wall or of its biosynthesis. In these studies, we have shown that chitin is a major structural component of G. lamblia and G. muris cyst walls. This conclusion is based on the finding that chitinase specifically destroys the cyst wall, as revealed by electron microscopy. The presence of chitin was also shown directly by lectin binding studies. Of 12 lectins with diverse carbohydrate recognition specificity, only the N-acetylglucosamine-specific lectins wheat germ agglutinin, succinylated wheat germ agglutinin, and tomato lectin bound to cyst walls, as shown by fluorescence microscopy and cytochemistry. Wheat germ agglutinin binding was completely abolished by treatment of the cysts with purified chitinase. This effect was specific since it could be prevented by incubating the enzyme with chitin before treatment of the cysts. Treatment of cysts with N-acetyl-beta-glucosaminidase partially inhibited wheat germ agglutinin binding, whereas other glycosidases and proteases had no effect. These findings indicate that chitin is a major structural component of Giardia cyst walls and raise the possibility that inhibitors of chitin synthesis may be of use in preventing encystation and thus controlling spread of the disease.
Ecology of Candida albicans gut colonization: inhibition of Candida adhesion, colonization, and dissemination from the gastrointestinal tract by bacterial antagonism
Authors: Kennedy M J , Volz P A
Published: January 2020

Antibiotic-treated and untreated Syrian hamsters were inoculated intragastrically with Candida albicans to determine whether C. albicans could opportunistically colonize the gastrointestinal tract and disseminate to visceral organs. Antibiotic treatment decreased the total population levels of the indigenous bacterial flora and predisposed hamsters to gastrointestinal overgrowth and subsequent systemic dissemination by C. albicans in 86% of the animals. Both control hamsters not given antibiotics and antibiotic-treated animals reconventionalized with an indigenous microflora showed significantly lower gut populations of C. albicans, and C. albicans organisms were cultured from the visceral organs of 0 and 10% of the animals, respectively. Conversely, non-antibiotic-treated hamsters inoculated repeatedly with C. albicans had high numbers of C. albicans in the gut, and viable C. albicans was recovered from the visceral organs of 53% of the animals. Examination of the mucosal surfaces from test and control animals indicated further that animals which contained a complex indigenous microflora had significantly lower numbers of C. albicans associated with their gut walls than did antibiotic-treated animals. The ability of C. albicans to associate with intestinal mucosal surfaces also was tested by an in vitro adhesion assay. The results indicate that the indigenous microflora reduced the mucosal association of C. albicans by forming a dense layer of bacteria in the mucus gel, out-competing yeast cells for adhesion sites, and producing inhibitor substances (possibly volatile fatty acids, secondary bile acids, or both) that reduced C. albicans adhesion. It is suggested, therefore, that the indigenous intestinal microflora suppresses C. albicans colonization and dissemination from the gut by inhibiting Candida-mucosal association and reducing C. albicans population levels in the gut.
Soluble non-cross-linked peptidoglycan polymers stimulate monocyte-macrophage inflammatory functions
Authors: Gold M R , Miller C L , Mishell R I
Published: January 2020

Soluble non-cross-linked peptidoglycan polymers are released by gram-positive bacteria when beta-lactam antibiotics are administered to humans. In this report, we show that this type of peptidoglycan can stimulate monocyte-macrophage functions that cause inflammation. Non-cross-linked peptidoglycan polymers from penicillin-treated Streptococcus faecium were purified and shown to stimulate the production of interleukin 1 by human monocytes and of colony-stimulating factors by a murine macrophage cell line. In addition, the release of plasminogen activator by human monocytes was inhibited by the soluble peptidoglycan. These in vitro results suggest that prolonged treatment with beta-lactam antibiotics, by causing the production of soluble peptidoglycan, may result in interleukin 1-mediated inflammatory reactions, excessive production of monocytes and granulocytes, and increased fibrin deposition.