bluuebunny/crossref_metadata_2025 huggingface dataset:
Protection of mice against Vibrio vulnificus disease by vaccination with surface antigen preparations and anti-surface antigen antisera
Authors: Kreger A S , Gray L D , Testa J
Published: January 2020

Vaccination of mice with either Formalin-killed cells or cell extracts of a virulent strain and a weakly virulent strain of Vibrio vulnificus or with rabbit antisera (AS) against the Formalin-killed cells and cell extracts protected against the virulent strain of V. vulnificus. However, the virulent strain vaccines and AS elicited a significantly stronger immune response than the weakly virulent strain vaccines and AS. Adsorption of the AS with either the homologous or heterologous Formalin-killed cells significantly reduced the ability of the AS to protect mice. The major protective antigen(s) in the cell extracts migrated in the void volume of Sephacryl S-400 superfine, was not effectively sedimented by centrifugation at 100,000 X g for 2 h, had an isoelectric point of 3.8 to 4.2, and was sensitive to boiling or autoclaving for 15 min, periodate oxidation, and exposure to pH 12 but was resistant to 56 degrees C, trypsin, pronase, RNase, neuraminidase, and pH 4.5. Electron microscopy revealed that the virulent strain possessed a more dense ruthenium red-staining layer on its outer membrane and had a much smoother surface than the weakly virulent strain. Our results provide evidence that a major protective antigen and virulence factor of V. vulnificus is a heat-labile, acidic polysaccharide located on the bacterial surface.
Staphylococcal toxic shock toxin specifically binds to cultured human epithelial cells and is rapidly internalized
Authors: Kushnaryov V M , MacDonald H S , Reiser R , Bergdoll M S
Published: January 2020

Staphylococcal toxic shock syndrome toxin (TST) was labeled with 125I under mild conditions without apparent destruction of the molecule. [125I]TST bound specifically to human epithelial (Chang) cells in culture; the binding was inhibited by a 100-fold excess of unlabeled toxin. Scatchard analysis of the binding data indicated about 10(4) receptor sites per cell and a dissociation constant (Kd) of 4 X 10(-9) M. When cells pretreated with TST at 4 degrees C were swiftly transferred to 37 degrees C, the amount of surface-bound toxin rapidly declined, as determined by release of noninternalized label from the cell surface. Half-time (t1/2) of internalization was about 1.5 min. Ultrastructural studies showed that toxin labeled with ferritin-conjugated antibodies entered the cytoplasm via coated pits forming coated vesicles in the first 2 min of incubation at 37 degrees C. The coated vesicles coalesced with transport vesicles that are ultrastructurally unlike receptosomes. Thus, the unusual ultrastructural pattern of this internalization suggests that TST is initially internalized by receptor-mediated endocytosis and then enters an alternate pathway involving translocation in special transport vesicles, perhaps to other cells.
Serum sensitivity of a Pseudomonas aeruginosa mucoid strain
Authors: Schiller N L , Alazard M J , Borowski R S
Published: January 2020

The susceptibility of Pseudomonas aeruginosa 144M (a mucoid strain isolated from the sputum of a cystic fibrosis patient) to the bactericidal activity of pooled fresh normal human serum (FHS) was examined. FHS at concentrations of greater than or equal to 2.5% was capable of killing greater than 95% of strain 144M. Strain 144M was killed by FHS in a dose-dependent manner. Although either immunoglobulin M (IgM) or IgG was bactericidal in the presence of complement, IgM was about 10 times as effective as IgG. However, optimal killing activity required both IgM and IgG and complement, activated by the classical pathway. A role for lysozyme in the killing of 144M was demonstrated only when low concentrations of FHS were used. In contrast to 144M, P. aeruginosa strains 144NM and 144M(SR) were totally resistant to FHS at all of the concentrations tested (up to 50%). Neither the FHS susceptibility of 144M nor the FHS resistance of 144NM or 144M(SR) was altered by choice of growth medium, growth phase, or temperature of growth. Results of absorption studies with whole organisms, isolated outer membrane preparations, or lipopolysaccharide (LPS) from each strain suggest that the antigen(s) which binds the bactericidal immunoglobulins is accessible on the surface of 144M but not on the surface of 144NM or 144M(SR), is insensitive to trypsin treatment, and is believed to be LPS. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the three LPS preparations demonstrated that 144M LPS contained primarily lipid-A-core polysaccharide components, whereas the LPS from 144NM and 144M(SR) were heterogeneous, with various degrees of O-side-chain substitution. These results suggest that at least one target for bactericidal antibody on the surface of 144M is contained in the rough LPS of this strain.
Weak serum and intestinal antibody responses to Vibrio cholerae soluble hemagglutinin in cholera patients
Authors: Svennerholm A M , Levine M M , Holmgren J
Published: January 2020

A soluble hemagglutinin/protease from Vibrio cholerae has been suggested to be a putative virulence factor and protective antigen. However, clinical cholera infection gave rise to detectable serum antibody responses to soluble hemagglutinin in only 2 of 10 Bangladeshi patients or 1 of 17 cholera-infected North American volunteers. A gut mucosal immunoglobulin A antibody response to soluble hemagglutinin was seen in 4 of 8 Bangladeshi patients, but in 0 of 10 North American volunteers. These responses were much weaker than those to cholera toxin or lipopolysaccharide.
Antigenic cross-reactivity between Treponema pallidum and other pathogenic members of the family Spirochaetaceae
Authors: Baker-Zander S A , Lukehart S A
Published: January 2020

The antigenic cross-reactivity between Treponema pallidum and several pathogenic members of the family Spirochaetaceae was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting techniques. Blots of T. pallidum antigens were incubated with antiserum from rabbits infected or immunized with T. pallidum, Treponema paraluiscuniculi, Treponema hyodysenteriae (strains B204 and T22), Borrelia hermsii serotype 7, or Leptopsira interrogans serogroup Canicola. T. pallidum contained 22 antigenic molecules ranging from 85,000 to 12,000 daltons which were recognized by serum from rabbits infected with T. pallidum. Serum from rabbits infected with T. paraluiscuniculi cross-reacted with 21 of these molecules and faintly reacted with a band at 15,000 daltons which was not recognized by anti-T. pallidum serum. Antisera directed against strains B204 and T22 of T. hyodysenteriae cross-reacted with 11 and 10 antigens of T. pallidum, respectively. B. hermsii and L. interrogans serogroup Canicola antisera detected 11 and 10 treponemal antigens, respectively. Many of the T. pallidum antigens detected by antisera against T. hyodysenteriae, B. hermsii, or L. interrogans serogroup Canicola have been previously identified as containing moieties also found on the nonpathogenic Treponema phagedenis, biotype Reiter, and may therefore represent group antigens common to members of the family Spirochaetaceae.
Experimental Lyme disease in rabbits: spirochetes found in erythema migrans and blood
Authors: Kornblatt A N , Steere A C , Brownstein D G
Published: January 2020

In attempts to produce experimental Lyme disease, 33 rabbits were inoculated with Lyme spirochetes by tick feeding or from tick organ homogenates or cultures. Two rabbits developed erythema chronicum migrans at the site of inoculation, in one instance 2 days after injection of a tick organ homogenate and in the other instance, 17 days after feeding of infected Ixodes dammini ticks. Spirochetes were seen in skin biopsy specimens of the second lesion with Warthin-Starry and immunoperoxidase stains. Spirochetes were also recovered from blood cultures of two additional rabbits 2 weeks post-inoculation. These findings are characteristic of early Lyme disease in humans and give additional support for the spirochetal etiology of Lyme disease.
Comparison of effects of nicked and unnicked Escherichia coli heat-labile enterotoxin on Chinese hamster ovary cells
Authors: Tsuji T , Honda T , Miwatani T
Published: January 2020

The effect on nicking (proteolytic cleavage) of heat-labile enterotoxin (LT) on its activities on Chinese hamster ovary (CHO) cells was examined. LT nicked with trypsin induced morphological change (elongation) of CHO cells from round to spindle-shaped cells faster than did unnicked LT. The times for elongation of 50% of the CHO cells with nicked and unnicked LT were 3 and 6 h, respectively. The activity of nicked LT on CHO cells was not inhibited by antiserum 20 min after treatment, whereas that of unnicked LT was inhibited by antiserum until ca. 130 to 160 min after treatment. These data suggest that unnicked LT remains on the cell surface longer than nicked LT and thus induces morphological changes of CHO cells more slowly than does nicked LT.
Production and characterization of monoclonal antibodies to Rickettsia rickettsii
Authors: Lange J V , Walker D H
Published: January 2020

Five mouse ascitic fluids (MAFs) containing monoclonal antibody to Rickettsia rickettsii were produced from three original fusions by murine hybridoma technology. The five MAFs were fractionated and purified; each contained monoclonal antibody of the immunoglobulin G2a subclass. Each monoclonal antibody-containing MAF was titrated by indirect immunofluorescence against three R. rickettsii isolates from humans and four other spotted fever group rickettsiae. Each MAF was also titrated in the complement fixation, latex agglutination, microagglutination, and indirect hemagglutination tests. Two of the MAFs were examined for their ability to prevent fever and rickettsemia in susceptible guinea pigs after a 1:100 dilution of each was mixed with viable R. rickettsii, and all five MAFs were titrated in the mouse toxicity phenomenon assay. All MAFs had high indirect immunofluorescence titers to the three strains of R. rickettsii (1:200,000 to 1:800,000), reduced indirect immunofluorescence titers to R. montana, and were nonreactive with R. akari, R. sibirica, and R. conorii. Each MAF was able to fix complement in the presence of spotted fever group antigen reagent and agglutinate a suspension of purified R. rickettsii, and each was negative in both the latex agglutination and the indirect hemagglutination tests. The two MAFs which were tested proved to be capable of preventing rickettsemia and death in guinea pigs, and each MAF was able to prevent death in mice at dilutions ranging from 1:40 to 1:80.
Antigenic S-type lipopolysaccharide of Brucella abortus 1119-3
Authors: Caroff M , Bundle D R , Perry M B , Cherwonogrodzky J W , Duncan J R
Published: January 2020

Antigenic phenol-phase soluble lipopolysaccharide isolated from Brucella abortus 1119-3 by hot phenol-water extraction was shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, controlled hydrolysis, periodate oxidation, methylation, and 1H and 13C nuclear magnetic resonance studies to be an S-type lipopolysaccharide which could be cleaved to yield a lipid A and an O-chain polysaccharide identified as an unbranched linear homopolymer of 1,2-linked 4,6-dideoxy-4-formamido-alpha-D-mannopyranosyl residues. The serological reactivity of bovine antiserum to B. abortus 1119-3 with the lipopolysaccharides of Yersinia enterocolitica serotype O:9 and Vibrio cholerae species has now been related to the occurrence of 1,2-linked N-acylated 4-amino-4,6-dideoxy-alpha-D-mannopyranosyl units in the O-chain polysaccharides of their lipopolysaccharides.
Exclusive presence of lactose-sensitive fimbriae on a typical strain (WVU45) of Actinomyces naeslundii
Authors: Cisar J O , David V A , Curl S H , Vatter A E
Published: January 2020

Lactose-sensitive fimbriae were identified as the only fimbriae present on Actinomyces naeslundii WVU45 (ATCC 12104). A single antigen reactive with antiserum against WVU45 cells was detected by cross immunoelectrophoresis of isolated fimbriae, and a monospecific antiserum against this antigen reacted with all fimbriae observed on the bacterial surface by immunoelectron microscopy. Moreover, the loss of one cell surface antigen by a spontaneous mutant of A. naeslundii WVU45 (WVU45M), isolated by its failure to react with a monospecific antibody against the fimbriae, was associated with the loss of all fimbriae. The functional involvement of the fimbriae in lactose-sensitive bacterial adherence was demonstrated by the ability of WVU45, but not WVU45M, cells to agglutinate neuraminidase-treated erythrocytes and by the lactose-sensitive hemagglutinating activity of immune complexes formed with isolated fimbriae and monospecific antibody. Bacterial agglutination assays with different monospecific antibodies revealed an antigenic similarity between the fimbriae of A. naeslundii WVU45 and the lactose-sensitive fimbriae (type 2) of Actinomyces viscosus T14V. In contrast, cross-reactivity was not observed between the WVU45 fimbriae and type 1 fimbriae, the structures involved in lactose-resistant adherence of strain T14V to saliva-treated hydroxyapatite. Functional differences between the fimbriae of A. naeslundii and A. viscosus strains may be correlated with well-established differences in the in vivo distribution of these organisms: namely, the preference of typical A. naeslundii for epithelial surfaces and of A. viscosus for tooth surfaces.
In vitro neutralization of Chlamydia trachomatis with monoclonal antibody to an epitope on the major outer membrane protein
Authors: Peeling R , Maclean I W , Brunham R C
Published: January 2020

A murine monoclonal antibody, which binds to an epitope on the major outer membrane protein of Chlamydia trachomatis and with species specificity in the micro-immunofluorescent assay, effectively neutralized in vitro two antigenically distinct serovars of C. trachomatis. Optimal concentrations of both organism and antibody were required to produce maximal neutralization of the organism. Neutralization was less effective and more variable at lower dilutions of antibody than at higher dilutions, suggesting a prozone phenomenon. A radiolabeled attachment assay demonstrated that attachment of elementary bodies was unaffected by earlier treatment with antibody and that neutralization occurred at a step after attachment. The epitope to which this antibody is directed, on the major outer membrane protein of C. trachomatis, may have an important role in determining infectivity of the organism.
Inhibition of phagocytosis and chemiluminescence in human leukocytes by a lipid soluble factor in normal tissues
Authors: Huang T S , Hurd R E , Chopra I J , Stevens P , Solomon D H , Young L S
Published: January 2020

Homogenates of normal rat tissues inhibited several functional parameters of normal human peripheral blood leukocytes, including luminol-dependent chemiluminescence induced by both soluble (phorbol myristate acetate) and particulate (Escherichia coli) stimuli; in vitro uptake of radiolabeled E. coli; and in vitro phagocytosis and killing of E. coli. The doses of rat tissue protein that caused a 50% inhibition of leukocyte chemiluminescence were ca. 6.2 micrograms for small intestine, 83 micrograms for kidney; 100 micrograms for heart; 132 micrograms for liver, 190 micrograms for skeletal muscle, and 307 micrograms for brain. The putative phagocytosis inhibitor (PI) in rat liver was more plentiful in particulate fractions than in the cytosol. The PI activity in the original or Miranol-solubilized rat liver homogenate was nondialyzable, and it was reduced substantially by heating at 90 degrees C for 30 min but not at 56 degrees C for 30 min. It was unaffected by aprotinin, a potent inhibitor of proteolytic activity. Treatment of tissues with trypsin did not reduce PI activity, whereas treatment with phospholipase A2 clearly increased it. The bulk (up to 88%) of PI in rat liver or small intestine could be extracted by lipid solvents, e.g., diethyl ether. Purified fatty acids were potent inhibitors of leukocyte chemiluminescence; other lipids had little or no inhibiting activity. The various data suggest that (i) normal tissues contain a potent PI and (ii) that the PI is a lipid moiety.
Conservation of plasmids among Escherichia coli K1 isolates of diverse origins
Authors: Mercer A A , Morelli G , Heuzenroeder M , Kamke M , Achtman M
Published: January 2020

Escherichia coli K1 isolates of various O types were previously assigned to different clonal groups. Members of the two clones defined by membrane pattern 9 (MP9) and serotypes O18:K1 and O1:K1 had been found to be very similar to each other. The plasmid contents of these bacteria confirmed this conclusion. Both groups carried a self-transmissible plasmid of the FI incompatibility group that coded for colicin production and a major outer membrane protein called the plasmid-coded protein (PCP). The size of this plasmid varied from 76 to 96 megadaltons, but restriction endonuclease digestion and DNA heteroduplex analysis revealed that these plasmids were highly related. O18:K1 bacteria of MP6 had previously been determined to represent a subclone, related to but different from O18:K1 MP9 bacteria. These MP6 bacteria carried a different, smaller IncFI plasmid which did not code for colicin production or the PCP protein. This smaller plasmid was primarily related to the larger plasmid within the regions of DNA encoding incompatibility, replication, and conjugation. O1:K1 bacteria of MP5 contained other unrelated plasmids in agreement with the previous conclusion that they are unrelated to O1:K1 bacteria of MP9. The bacteria examined had been isolated from two continents over a time span of 38 years, and the results attest to conservative inheritance of plasmids within bacteria of common descent.
Pathogenicity of attaching effacing enteropathogenic Escherichia coli isolated from diarrheic suckling and weanling rabbits for newborn rabbits
Authors: Peeters J E , Charlier G J , Halen P H
Published: January 2020

The pathogenicity of six strains of Escherichia coli originating from different commercial rabbitries was tested in neonatal rabbits. Two strains isolated from healthy weaned rabbits (O7:H6 and O9:H?) did not induce any clinical sign or lesion. Two strains (O109:H2) isolated from diarrheic suckling rabbits caused yellow diarrhea 36 to 60 h after inoculation and high mortality between 60 and 72 h after infection. At 12 h after infection, light and electron microscopy showed attachment to epithelial cells and effacement of microvilli from proximal small intestine to colon. Bacteria were often present in the apical cytoplasm of epithelial cells. The two strains isolated from diarrheic weanling rabbits (O109:H2 and O15:H-) did not induce any clinical sign. Attachment to epithelial cells and effacement of microvilli was observed 48 h after inoculation in distal small intestine, cecum, and colon. These data are further evidence for the existence of two groups of attaching effacing enteropathogenic E. coli in rabbits, showing different preferences for age group and intestinal compartment.
Botulism in metronidazole- treated conventional adult mice challenged orogastrically with spores of Clostridium botulinum type A or B
Authors: Wang Y , Sugiyama H
Published: January 2020

Conventional adult mice were pretreated with metronidazole to make their intestinal tracts receptive to colonization by Clostridium botulinum. These mice, in groups of 10, were fed 0 (controls), 10(2), 10(3), 10(4), or 10(5) C. botulinum type B spores and were placed for observation in filter-lid cages whose screen floors minimized the amounts of feces available for coprophagy. The opportunity to eat feces was made equal for all mouse groups by placing one mouse of every group in each of 10 cages. Mice given a spore inoculum began to develop botulism after incubation periods of slightly less than 2.75 days. Morbidity rates, which reached maxima within 5 days of challenge, were related to inocula levels. Mortality rates were also dose related. Mice given 10(5) spores and then type B antitoxin intraperitoneally, a treatment not affecting intraintestinal toxin production, remained healthy. Morbidity among control mice was seldom more than 10% and could be ascribed to toxin ingested with feces. A C. botulinum type A spore suspension gave similar results, although morbidity and mortality rates were generally lower than after challenge with a comparable number of type B spores. Mice challenged with 10(2) or 10(5) spores had similar toxin levels in their large intestines 48 h later. Morbidity rates correlated better with toxin levels in the small intestines.
A plasmid-encoded outer membrane protein, TraT, enhances resistance of Escherichia coli to phagocytosis
Authors: Agüero M E , Aron L , DeLuca A G , Timmis K N , Cabello F C
Published: January 2020

The presence of the outer membrane protein TraT, encoded by plasmid R6-5, reduces the sensitivity of Escherichia coli cells to phagocytosis by macrophages. This effect is independent of the bacterial capsule and is more evident in the presence of adsorbed normal human serum. The property of inhibiting phagocytosis is specifically abolished by anti-TraT protein antiserum and anti-TraT immunoglobulin G but not by Fab fragments. These results indicate that the TraT protein is a passive inhibitor of phagocytosis. Inhibition of phagocytosis is produced because the TraT protein antagonizes opsonization by complement, such that C3 deposition is reduced and altered in distribution.
Effect of antigen form on local immunoglobulin A memory response of intestinal secretions to Shigella flexneri
Authors: Keren D F , McDonald R A , Scott P J , Rosner A M , Strubel E
Published: January 2020

An enhanced memory response, as shown by increased titers of specific immunoglobulin A (IgA), was seen in intestinal secretions from isolated Thiry-Vella loops in rabbits primed orally with live, locally invasive Shigella sp. X16 and challenged 60 days later with a single oral dose of the same antigen. Heat-killed shigella preparations, when used as either the priming or challenge antigen, did not elicit such a memory response in this system. In the present study, the role of antigen form and dosage in eliciting the enhanced local IgA response was investigated. A noninvasive strain, Shigella flexneri 2457-0, was capable of significantly enhancing the mucosal IgA memory response, whereas heat-killed Shigella sp. X16 was unable to augment the local IgA response, even when the priming dose was increased 100-fold. A proposed mucosal adjuvant, DEAE-dextran, given orally with live Shigella sp. X16, did not enhance the local IgA response. Viable, noninvasive shigellae were effective priming agents in enhancing the local IgA memory response. The poor mucosal response to heat-killed shigella preparations is thought to be related to an ineffective delivery of nonviable bacterial antigens into gut-associated lymphoid tissues. The ability of the live, noninvasive strain to elicit a vigorous local IgA memory response when given orally to rabbits was consistent with previous findings that live preparations elicit the best mucosal IgA response.
Suppression of in vitro lymphocyte proliferation in C57BL/10 ScN mice vaccinated with phase I Coxiella burnetii
Authors: Damrow T A , Williams J C , Waag D M
Published: January 2020

The effect of inactivated phase I and phase II Coxiella burnetii whole cell vaccine (WCV) on the response of murine spleen cells to mitogenic and antigenic stimuli was evaluated in C57BL/10 ScN endotoxin nonresponder mice with an in vitro lymphocyte proliferation assay. Intraperitoneal injection of phase I WCV into mice resulted in marked and persistent suppression of the proliferative response of spleen cells to concanavalin A, phytohemagglutinin, and pokeweed mitogen. This response was time and dose dependent and was not associated with decreased lymphocyte viability. By using a standard dose of 100 micrograms of phase I WCV, suppression of mitogenic responsiveness was first detected 3 days postinjection, attained maximum levels by day 14, and persisted for longer than 5 weeks. Suppression of mitogenic lymphocyte proliferation also was demonstrated after inoculation of animals with viable phase I organisms. The observed hyporesponsiveness of spleen cells from phase I WCV-injected animals was not either the result of a shift in the mitogenic dose optimum or due to a change in the day of in vitro peak response. Spleen cells from phase I WCV-injected mice were negatively regulated with homologous antigen. Investigation of the mechanism of action of phase I WCV, with a 51Cr-release assay, and trypan blue dye exclusion showed that phase I WCV was not directly cytolytic or cytotoxic to spleen cells from normal or vaccinated mice. Phase II WCV did not induce significant mitogenic hyporesponsiveness or negative modulation of spleen cells. These findings extend the observations of adverse host responses associated with the phase I WCV and underscore the need to develop a microbial fraction which possesses protective potency but which lacks the propensity to induce deleterious tissue reactions and immunosuppression.
Lipopolysaccharide characteristics of pathogenic campylobacters
Authors: Perez Perez G I , Blaser M J
Published: January 2020

Most Campylobacter jejuni strains are sensitive and most Campylobacter fetus strains are resistant to the bactericidal activity in normal human serum. We purified lipopolysaccharides from Campylobacter strains to determine whether their composition and structure relate to serum susceptibility. The lipopolysaccharide of two serum-sensitive strains was best isolated by the Galanos procedure, but for two serum-resistant strains a cold-ethanol extraction was optimal. For each lipopolysaccharide preparation, the ratio of 2-keto-3-deoxyoctonate to protein was increased by 100 to 1,000-fold over that of whole cells. For serum-resistant strains, total carbohydrates was a high proportion of lipopolysaccharide weight; for serum-sensitive strains, 2-keto-3-deoxyoctanate was a high proportion of total carbohydrates. By polyacrylamide gel electrophoresis, the lipopolysaccharide of serum-sensitive strains appeared rough, but for serum-resistant strains a smooth-type ladder was seen, with a minimal core region and several high-molecular-weight complexes. Proteinase K-treated whole-cell lysates showed polyacrylamide gel electrophoresis profiles similar to that of pure lipopolysaccharide. Proteinase K-treated whole-cell lysates from seven serum-sensitive C. jejuni strains all had rough profiles, and five serum-resistant C. fetus strains all had smooth profiles. These studies indicate that lipopolysaccharide composition may be an important determinant of serum susceptibility among Campylobacter species and that serum resistance is usually associated with a smooth-type lipopolysaccharide.
Candidacidal activity of myeloperoxidase: therapeutic influence of the enzyme in vivo
Authors: Wright C D , Nelson R D
Published: January 2020

Mice were injected intravenously with Candida albicans blastospores to establish chronic renal infection. Seventy-five percent of the animals inoculated with 10(6) blastospores died as a consequence of infection during the subsequent 60 days of observation. Intraperitoneal administration of 10 mU of partially purified human myeloperoxidase 1 day after injection of the pathogen increased survival of the mice from 25 to 80% over this time period. Administration of myeloperoxidase complexed with soluble C. albicans cell wall mannan abrogated the protective influence of the enzyme. These results demonstrate that exogenous myeloperoxidase has a therapeutic influence on murine renal candidiasis and suggest that exogenous myeloperoxidase may also be effective in the treatment of certain forms of candidiasis in humans. These results also demonstrate the important role of the mannan-binding function of myeloperoxidase for effective treatment of candidiasis and suggest a mechanism of inhibition of the candidacidal effect of free enzyme in vivo by mannan accumulating in tissue fluids.
Relationship between extracellular stimulation of intracellular killing and oxygen-dependent microbicidal systems of monocytes
Authors: Leijh P C , Nathan C F , van den Barselaar M T , van Furth R
Published: January 2020

Human monocytes require serum components immunoglobulin G, C3/C3b, and B/Bb to exert optimal microbicidal action against ingested microorganisms. The present study was performed to find out whether these factors act by enhancing oxygen-dependent antimicrobial mechanisms. Serum enhanced oxygen consumption and superoxide production by monocytes before phagocytosis, but did not further increase these processes in monocytes that had recently ingested bacteria. Furthermore, serum did not boost iodination during intracellular killing by monocytes. Phorbol myristate acetate, N-formyl-methyonyl-leucyl-phenylalanine, concanavalin A, and concanavalin A-Sephadex all stimulated the conversion of O2 to H2O2 by monocytes, but only concanavalin A augmented intracellular killing. Reactive oxygen intermediates generated by cell-free enzymes (xanthine oxidase or glucose oxidase) in concentrations comparable to those accumulating extracellularly during incubation of monocytes containing bacteria with phorbol myristate acetate did not promote intracellular killing. The presence of catalase during phagocytosis inhibited killing, but had no effect on killing in the postphagocytic state. Monocytes deprived of glucose for 24 h showed markedly impaired O2 consumption, O2- generation, and bacterial killing; all of these effects were rapidly reversed by restoration of glucose. It is concluded that both an intact respiratory burst and extracellular serum factors are necessary for optimal killing of intracellular Staphylococcus aureus by human monocytes. Serum does not appear to act by enhancing the respiratory burst, but rather to have a separate, synergistic role, the biochemical basis of which is unknown.
Differential inhibition of macrophage microbicidal activity by liposomes
Authors: Gilbreath M J , Swartz G M , Alving C R , Nacy C A , Hoover D L , Meltzer M S
Published: January 2020

In vitro culture of murine resident peritoneal macrophages with lymphokine (LK)-rich leukocyte culture fluids induces enhanced microbicidal activity against amastigotes of the protozoan parasite Leishmania tropica. Macrophages infected with Leishmania and treated with LKs after infection acquire the capacity to kill the intracellular parasite within 72 h. When compared with control macrophage cultures treated with medium lacking LKs, 80 to 90% fewer macrophages treated with LKs contained amastigotes. In experiments designed to test liposome delivery of LKs to infected macrophages, addition of multilamellar liposomes composed of phosphatidylcholine and phosphatidylserine (molar ratio, 7:3) completely abrogated LK-induced microbicidal activity. Liposomes containing only phosphatidylcholine were not inhibitory. Inhibition of LK activity by the liposomes occurred regardless of whether the liposomes contained LKs. Liposomal inhibition of activated macrophage effector activity was limited to intracellular killing; LK-induced macrophage extracellular cytolysis (i.e., tumor cytotoxicity) was not affected by liposome treatment. These data indicate that elucidation of the effects of liposome composition on acquired host defense mechanisms may be useful for the design of drug delivery systems that allow expression or augmentation of immunologically induced mechanisms for the intracellular destruction of infectious agents.
Blastogenic response of bovine lymphocytes to Brucella abortus lipopolysaccharide
Authors: Baldwin C L , Winter A J
Published: January 2020

Brucella abortus lipopolysaccharide was tested in a blastogenesis assay with unfractionated and nylon wool-separated peripheral blood lymphocytes of Brucella-naive cattle and cattle immunized with B. abortus. Our results indicated that in cattle the lipopolysaccharide of B. abortus is not a B-cell mitogen. In immunized animals it stimulated predominantly nylon wool-adherent cells. The lipopolysaccharide of Escherichia coli O128:B12, in contrast, induced a substantially greater proliferative response in circulating lymphocytes, predominantly those adherent to nylon wool, of the Brucella-naive cattle.
Immunity to Salmonella typhimurium infection in C3H/HeJ and C3H/HeNCrlBR mice: studies with an aromatic-dependent live S. typhimurium strain as a vaccine
Authors: Killar L M , Eisenstein T K
Published: January 2020

Immunization with avirulent Salmonella typhimurium strain SL3235, a smooth, aroA- derivative, was shown to induce high levels of resistance to challenge with virulent S. typhimurium in innately hypersusceptible C3H/HeJ mice and inherently resistant C3H/HeNCrlBR mice. Strain SL3235 is one of a class of avirulent aroA- derivatives made from various strains and species of Salmonella that are being considered as vaccine candidates for cattle and humans. This paper supports their efficacy and potential utility in this regard. In C3H/HeJ mice, immunity against over 1,000 50% lethal doses of virulent S. typhimurium was evident as early as 3 days after immunization and persisted for at least 7 months. Further, the vaccine was effective over a broad spectrum of doses, ranging from 10(4) to 10(6) organisms. Infection with SL3235 led to marked splenomegaly in both mouse strains. The relationship of splenomegaly to the growth kinetics and colonization by SL3235 in the spleens of infected C3H/HeJ and C3H/HeNCrlBR mice was followed. SL3235 initially multiplied slowly in the spleens of both mouse strains and then was rapidly cleared. Less multiplication was seen in the resistant C3H/HeNCrlBR mice than in C3H/HeJ mice. Maximum splenomegaly occurred after clearance of the organism had begun. Protection against virulent S. typhimurium persisted after virtually all of the SL3235 vaccine strain had been cleared from the spleen. Cross-protection against Listeria monocytogenes was evident, but had a later onset, waned by 21 days, and was not detectable by 1 month after vaccination. Demonstration of this cross-protection is consistent with the interpretation that SL3235 induces cellular immunity. One-week immune spleen cells adoptively transferred anti-S. typhimurium and anti-L. monocytogenes immunity. T cell-enriched fractions were ineffective in adoptive transfer, as were spleen cells taken 2 weeks or later after immunization. Protective capacity was in the adherent cell fraction and seemed to be associated with macrophages. Evidence for induction of a population of sensitized T cells was obtained by using a peritoneal exudate T-lymphocyte proliferation assay on peritoneal T lymphocytes collected 1 to 3 months after SL3235 infection.
Identification of a 68-kilodalton protective protein antigen from Bordetella bronchiseptica
Authors: Montaraz J A , Novotny P , Ivanyi J
Published: January 2020

A 68-kilodalton (kd) outer membrane protein antigen of Bordetella bronchiseptica has been identified by using monoclonal antibodies that recognized two nonoverlapping determinants. Antibody BB05 also reacted with homologous proteins from Bordetella pertussis and Bordetella parapertussis but not with another 12 organisms from various bacterial genera. Passive injection of BB05 antibody protected mice from aerosol infection with B. bronchiseptica as shown by reduced mortality and reduced pathology of turbinate bones. The 68-kd B. bronchiseptica antigen was purified by BB05-based affinity chromatography and evaluated for its potency to immunize mice actively against either intraperitoneal or aerosol challenge with B. bronchiseptica. Immunization with the 68-kd antigen in incomplete Freund adjuvant significantly reduced the levels of mortality in intraperitoneally challenged mice. In the aerosol infection model, injection of the 68-kd antigen with complete or incomplete Freund adjuvant or saponin reduced the bacterial counts in the lungs of infected mice. These results suggest that the 68-kd protein may represent a potential "protective" antigen of B. bronchiseptica.
Effects of iron and desferrioxamine on infections with Yersinia enterocolitica
Authors: Robins-Browne R M , Prpic J K
Published: January 2020

The effects of iron-dextran and the iron chelator desferrioxamine B mesylate (Desferal) on the course and outcome of experimental yersiniosis were investigated. Yersinia enterocolitica strains representing the three leading serogroups pathogenic for humans, O3, O8 and O9, were studied. In mice, iron-dextran reduced the median lethal dose of intraperitoneally administered Y. enterocolitica O3 and O9 ca. 10-fold, whereas Desferal reduced this value more than 100,000-fold. Experiments in which Y. enterocolitica was given orally to mice and intraconjunctivally to guinea pigs confirmed that Desferal markedly increased the susceptibility of animals to yersiniosis. Although serogroup O8 yersiniae were inherently more virulent for laboratory animals, they were less affected by Desferal than were O3 or O9 strains. In vitro experiments indicated that Desferal promoted growth of Y. enterocolitica under iron-limiting conditions and suggested that the enhanced virulence of O8 yersiniae may be due to their comparatively low requirement for iron. The adverse effect of Desferal on the course of experimental infection with Y. enterocolitica may partly explain the heightened susceptibility of iron-overloaded patients to systemic yersiniosis.
Generation of oxygen species and virulence of Listeria monocytogenes
Authors: Godfrey R W , Wilder M S
Published: January 2020

Listeria monocytogenes strains differing in virulence were examined for production of oxygen species. Virulent strains A4413 and JH-H exhibited maximal generation of hydrogen peroxide and superoxide, whereas the avirulent strain 9037-7R was least active. The virulence was confirmed by the in vitro assessment of the intracellular fate of the microorganisms in resident macrophages.
Large plasmids associated with virulence in Shigella species have a common function necessary for epithelial cell penetration
Authors: Watanabe H , Nakamura A
Published: January 2020

Large plasmids (120 to 140 megadaltons) associated with virulence of Shigella sonnei, S. flexneri 2a and S. dysenteriae 1 were transferred from each strain into Escherichia coli K-12 and avirulent S. flexneri 1b strains by ampicillin transposon (Tn1)-mediated conduction. Strains with the virulence plasmid could penetrate tissue culture cells irrespective of the original host of the plasmid.
Purification and characterization of an extracellular cytolysin produced by Vibrio vulnificus
Authors: Gray L D , Kreger A S
Published: January 2020

An extracellular cytolytic toxin produced by the halophilic bacterium Vibrio vulnificus was isolated free of detectable contamination with medium constituents and other bacterial products by sequential ammonium sulfate precipitation, gel filtration with Sephadex G-75, hydrophobic interaction chromatography with phenyl-Sepharose CL-4B, and isoelectric focusing in an ethylene glycol density gradient. The cytolysin is a heat-labile, hydrophobic protein that is inhibited by large amounts of cholesterol, is partially inactivated by proteases and trypan blue, has a molecular weight (estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by amino acid analysis) of ca. 56,000, and has an isoelectric point of ca. 7.1. The first 10 amino-terminal amino acid residues of the cytolysin are Gln-Glu-Tyr-Val-Pro-Ile-Val-Glu-Lys-Pro. Lysis of mouse erythrocytes by the purified cytolysin is a multi-hit, at least two-step process consisting of a temperature-independent, toxin-binding step, followed by a temperature-dependent, membrane-perturbation step(s). In addition to possessing cytolytic activity against erythrocytes from 17 animal species and against Chinese hamster ovary cells in tissue culture, the purified cytolysin preparation was lethal for mice (ca. 3 micrograms/kg, intravenous 50% lethal dose) and had vascular permeability factor activity in guinea pig skin.
Effect of fibronectin on adherence of Staphylococcus aureus to fibrin thrombi in vitro
Authors: Toy P T , Lai L W , Drake T A , Sande M A
Published: January 2020

Staphylococcus aureus binds to purified fibronectin in solution and may bind to fibronectin present in wound tissue. When incorporated into a solid fibrin thrombus, however, plasma fibronectin may fail to bind S. aureus, because the S. aureus-binding sites on fibronectin may be occupied by fibrin. Both S. aureus and fibrin bind to the same 27-kilodalton amino-terminal fragment of fibronectin. To determine whether fibronectin incorporated into fibrin still promotes the adherence of S. aureus, we clotted citrated normal plasma and fibronectin-depleted plasma onto petri dishes. We then measured bacterial adherence to these in vitro fibrin thrombi. We found that the adherence of five of seven S. aureus strains decreased significantly (by 26 to 58%) when fibronectin had been depleted from the fibrin thrombi. Adding fibronectin back reversed this decrease in adherence. The reversal was dose dependent; the increase was in proportion to the amount of fibronectin added back to the plasma. Bacteria known not to bind to fibronectin (Escherichia coli and Staphylococcus epidermidis) adhered 100-fold less than S. aureus, and their adherence was unaffected by the absence of fibronectin in the fibrin thrombus. We conclude that fibronectin incorporated into solid fibrin thrombi does mediate the adherence of most S. aureus strains to fibrin thrombi. Fibronectin may be an important molecule that mediates the adherence of S. aureus to fibrin in wounds.
Serum antibody reactive with predominant organisms in the subgingival flora of young adults with generalized severe periodontitis
Authors: Tew J G , Marshall D R , Moore W E , Best A M , Palcanis K G , Ranney R R
Published: January 2020

In the present study we sought to determine whether serum antibody was present against microorganisms which predominate in the subgingival flora of young adults with generalized severe periodontitis (SP). Subjects with SP were often seropositive for Eubacterium brachy, Fusobacterium nucleatum E3C22, and Peptostreptococcus micros, whereas subjects with juvenile periodontitis (JP) and subjects with healthy periodontium (HP) were not. Both SP and JP subjects were more frequently seropositive for Bacteroides gingivalis, F. nucleatum D52B16, and F. nucleatum E1D1 than were HP subjects. The data were most striking for B. gingivalis, for which both the incidence and the magnitude of specific antibody was clearly elevated for SP and JP subject groups. However, SP subjects generally had either a high antibody titer or no detectable titer. In contrast, JP and HP subjects generally had at least very small amounts of antibody. Except at very low levels of antibody, neither SP nor JP groups differed significantly from the HP group for antibody to Eubacterium nodatum, Bacteroides intermedius (homology group 4197 or 8944), or Lactobacillus minutus antibody. There was a high frequency of antibody to E. nodatum, with very high titers in all groups despite the fact that this organism is rarely found in HP subjects. For Eubacterium timidum, the JP group was clearly more frequently seropositive than the HP group. Despite high levels of L. minutus in subgingival flora, none of the 50 SP subjects had a detectable antibody titer, and only four of the HP and JP subjects had detectable antibody. These results indicate that many organisms in the subgingival flora elicit antibody responses. B. gingivalis is probably the best example among the species tested. However, some organisms that are present in high concentration, e.g., L. minutus, apparently fail to induce significant antibody responses.
Effect of systemic immunization on pulmonary clearance of Haemophilus influenzae type b
Authors: Toews G B , Hart D A , Hansen E J
Published: January 2020

The effect of systemic immunization on pulmonary clearance of Haemophilus influenzae type b (Hib) was studied in a mouse model system. Immunization of mice by intraperitoneal injection of viable Hib cells resulted in the appearance of Hib-directed antibodies in both serum and bronchoalveolar lavage fluid. The development of this Hib-directed antibody activity was associated with significant enhancement of early pulmonary clearance of Hib. Systemic immunization did not affect the recruitment of polymorphonuclear leukocytes to the alveoli, suggesting that the enhanced clearance of Hib observed in immunized animals was due to specific antibodies which promote either phagocytosis or extracellular killing of Hib. The spectrum of Hib-directed antibody specificities detected in sera from immunized animals was essentially identical to that detected in bronchoalveolar lavage fluids from these same animals. Similarly, intravenous administration of an immunoglobulin G monoclonal antibody specific for Hib lipopolysaccharide resulted in the subsequent appearance of this antibody in the alveolar spaces where it enhanced pulmonary clearance of Hib. This study shows that this mouse model system can be used to measure the effect of both active and passive immunization on the clearance of Hib from the lower respiratory tract.
Lipopolysaccharide variation in Coxiella burnetti: intrastrain heterogeneity in structure and antigenicity
Authors: Hackstadt T , Peacock M G , Hitchcock P J , Cole R L
Published: January 2020

We isolated lipopolysaccharides (LPSs) from phase variants of Coxiella burnetii Nine Mile and compared the isolated LPS and C. burnetii cells by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. The LPSs were found to be the predominant component which varied structurally and antigenically between virulent phase I and avirulent phase II. A comparison of techniques historically used to extract the phase I antigenic component revealed that the aqueous phase of phenol-water, trichloroacetic acid, and dimethyl sulfoxide extractions of phase I C. burnettii cells all contained phase I LPS, although the efficiency and specificity of extraction varied. Our studies provide additional evidence that phase variation in C. burnetii is analogous to the smooth-to-rough LPS variation of gram-negative enteric bacteria, with phase I LPS being equivalent to smooth LPS and phase II being equivalent to rough LPS. In addition, we identified a variant with a third LPS chemotype with appears to have a structural complexity intermediate to phase I and II LPSs. All three C. burnetii LPS contain a 2-keto-3-deoxyoctulosonic acid-like substance, heptose, and gel Limulus amoebocyte lysates in subnanogram amounts. The C. burnetii LPSs were nontoxic to chicken embryos at doses of over 80 micrograms per embryo, in contrast to Salmonella typhimurium smooth- and rough-type LPSs, which were toxic in nanogram amounts.