bluuebunny/crossref_metadata_2025 huggingface dataset:
Frequency and organization of papA homologous DNA sequences among uropathogenic digalactoside-binding Escherichia coli strains
Authors: Denich K , Craiu A , Rugo H , Muralidhar G , O'Hanley P
Published: January 2020

The frequency of selected papA DNA sequences among 89 digalactoside-binding, uropathogenic Escherichia coli strains was evaluated with 12 different synthetic 15-base probes corresponding to papA genes from four digalactoside-binding piliated recombinant strains (HU849, 201B, and 200A). The papA probes encode amino acids which are common at the carboxy terminus of all strains, adjacent to the proximal portion of the intramolecular disulfide loop of strain 210B, or predicted to constitute the type-specific epitope for each of the four recombinant strains or other epitopes of strain HU849. The presence among the strains of DNA sequence homology to the papA probes was determined by in situ colony hybridization. Hybridization data suggest that there is a high frequency of homologous papA DNA sequences corresponding to selected regions of the papA gene from strain HU849 among the clinical strains. The following nucleotide locations which encode portions of the mature HU849 PapA are detected in a high percentage (42 to 70%) of clinical isolates: 208 to 222, 310 to 324, 478 to 492, 517 to 531, 553 to 567, and 679 to 693. These sequences encode portions of the predicted protective, immunogenic, and/or antigenic epitopes of this PapA. The data also indicate considerable heterogeneity of papA sequences among the strains, especially in the region of nucleotide bases corresponding to positions 391 to 418. These oligonucleotides encode the predicted PapA type-specific immunogenic dominant epitope. Determination of the extent of genetic variability in the papA gene among digalactoside-binding strains will require more extensive DNA sequencing of prototypic papA genes, additional hybridization studies employing other papA gene oligonucleotide probes, and assessment of the different pap operons and their copy number in each strain.
A lipopolysaccharide (LPS)-resistant mutant isolated from a macrophagelike cell line, J774.1, exhibits an altered activated-macrophage phenotype in response to LPS
Authors: Amano F , Akamatsu Y
Published: January 2020

A bacterial lipopolysaccharide (LPS)-resistant mutant was isolated from murine macrophagelike cell line J774.1. The mutant showed selective resistance to LPS and lipid A and was almost 10(5)- to 10(6)-fold more resistant than the parent; it grew even in the presence of 1 mg of Escherichia coli O55:B5 LPS per liter, whereas the parent did not grow with less than 10 ng of LPS per milliliter. We next examined the mutant for activation of various functions of macrophages on LPS treatment. This LPS-resistant mutant secreted interleukin-1 and tumor necrosis factor almost as effectively as the parent did. The mutant cells also changed transiently from a round to a spread form; however, they became round again afterwards. The mutant cells secreted less arachidonic acid in response to LPS. These results also suggest that this LPS-resistant mutant responds to LPS and shows activation of some macrophage functions. However, this mutant did not exhibit elevation of O2- generation or H2O2 generation after LPS treatment. Also, treatment of the mutant cells with murine recombinant gamma interferon was partly able to correct the defect in O(2-)-generating activity in response to LPS, suggesting that this defect is probably due to some of the LPS signal pathways. This implies that there is some correlation between O2- metabolism in LPS-activated macrophages and decreases in cell growth and viability.
Iron limitation and the gamma interferon-mediated antihistoplasma state of murine macrophages
Authors: Lane T E , Wu-Hsieh B A , Howard D H
Published: January 2020

The zoopathogenic fungus Histoplasma capsulatum requires iron for growth. Intracellular growth of the fungus within mouse peritoneal macrophages is inhibited by recombinant murine gamma interferon (IFN-gamma). Such treatment of mouse peritoneal macrophages induces a marked downshift in transferrin receptors. We tested whether the antihistoplasma effect of IFN-gamma-treated macrophages is the result of iron deprivation. Treatment of mouse peritoneal macrophages with the intracellular iron chelator deferoxamine inhibits the intracellular growth of H. capsulatum. Exposure of macrophages to holotransferrin antagonizes the effect of both recombinant murine IFN-gamma and deferoxamine treatments. These results suggest that iron restriction may be one of the bases for the IFN-gamma-induced antihistoplasma effect of mouse macrophages.
Treponema pallidum subsp. pallidum has a single, circular chromosome with a size of approximately 900 kilobase pairs
Authors: Walker E M , Arnett J K , Heath J D , Norris S J
Published: January 2020

The genome size and chromosome conformation of Treponema pallidum subsp. pallidum, Nichols strain, were determined by contour-clamped homogeneous electric field electrophoresis, a pulsed-field gel electrophoresis technique. Digestion of T. pallidum subsp. pallidum DNA with the restriction endonucleases NotI and SpeI produced 12 and 26 fragments, respectively. Summation of the physical lengths of the fragments produced by NotI and SpeI cleavage yielded average sizes of 900 and 913 kbp, respectively, for the genome of T. pallidum subsp. pallidum. Contour-clamped homogeneous electric field electrophoresis of T. pallidum subsp. pallidum DNA exposed to 4 krads of gamma irradiation resolved a single band of 800 to 1,000 kbp; treatment of the DNA with 16 krads of gamma irradiation resulted in the production of smaller fragments, whereas untreated DNA did not migrate into the gels. The gamma irradiation results indicate that T. pallidum subsp. pallidum has a single, circular chromosome that was linearized at a dosage of 4 krads of gamma irradiation. The size estimate provided by restriction endonuclease digestion with NotI and SpeI shows that the genome of T. pallidum subsp. pallidum, at approximately 900 kbp, is considerably smaller than the 13,700-kbp genome size calculated from renaturation kinetics.
Effect of mycobacteria on sensitivity to the cytotoxic effects of tumor necrosis factor
Authors: Filley E A , Rook G A
Published: January 2020

Unlike Mycobacterium leprae, Mycobacterium tuberculosis is not found inside cells other than macrophages and polymorphonuclear cells in vivo, yet previous work has revealed that in vitro it readily enters all cell lines tested. Moreover, these cells are not killed by the intracellular mycobacteria. We report here that when fibroblasts take up live (but not killed) M. tuberculosis H37Rv, they develop greatly increased sensitivity to the toxic effects of tumor necrosis factor (TNF) whether the cell line is inherently sensitive to TNF or not. Ultrasonically disrupted M. tuberculosis also has this property. The increased sensitivity is seen in the absence of metabolic inhibitors, although addition of emetine, an inhibitor of protein synthesis, causes the effect to manifest itself earlier and at a lower concentration of TNF. In contrast, infection with Mycobacterium bovis bacillus Calmette-Guérin induces little or no increased sensitivity to TNF, whereas Mycobacterium avium and M. tuberculosis H37Ra have intermediate sensitivities. We discuss the possibility that virulent tuberculosis strains produce a factor which distorts the normal protective function of TNF, rendering it toxic to host tissues and leading to the classical immunopathology of tuberculous lesions.
Role of neutrophil receptors in opsonophagocytosis of coagulase-negative staphylococci
Authors: Schutze G E , Hall M A , Baker C J , Edwards M S
Published: January 2020

The role of neutrophil complement receptors in the opsonophagocytosis of 10 strains of coagulase-negative staphylococci was investigated. Polymorphonuclear leukocytes from adults as well as term and premature newborn infants were tested with normal human serum, adult hypogammaglobulinemic serum, and pooled premature infant serum in an opsonophagocytic assay. Neutrophils from premature infants demonstrated significantly lower killing capacity (62%) than neutrophils from adults (86%) or term infants (84%; P less than 0.02). Maximum inhibition of opsonophagocytosis by adult or infant neutrophils occurred with an FcIII receptor blockade (80%), whereas a blockade of complement receptors produced minimal inhibition. Opsonophagocytic activity for the coagulase-negative staphylococci was not influenced by the serum source but was influenced by reducing the serum concentration below 5%. Abrogation of the complement activity of normal human serum by heating or the addition of ethylenediamine tetraacetate reduced opsonophagocytosis by 100 and 96%, respectively, whereas selective inhibition of the classical complement pathway reduced opsonophagocytosis by only 40%. Thus, opsonophagocytosis of coagulase-negative staphylococci by human sera appears to be mediated primarily by neutrophil Fc receptors, but complement is also required. The inefficiency of these interactions with neutrophils from premature infants may partially explain the enhanced susceptibility of very-low-birth-weight neonates to disseminated, coagulase-negative staphylococcal infections.
Analysis of immune responses of different hosts to Babesia divergens isolates from different geographic areas and capacity of culture-derived exoantigens to induce efficient cross-protection
Authors: Precigout E , Gorenflot A , Valentin A , Bissuel G , Carcy B , Brasseur P , Moreau Y , Schrevel J
Published: January 2020

The immunoprecipitation of [35S]methionine-radiolabelled antigens from different Babesia divergens isolates by using bovine, gerbil, and human immune sera has shown that many B. divergens proteins contain epitopes shared between isolates. The cross-protective capacity of culture-derived soluble immunogens from the B. divergens Rouen 1987 isolate was tested against different B. divergens isolates. Results showed complete protection against the 7107b French isolate and substantial protection against the Weybridge 8843 English isolate (80% protection) and the Munich 87 German isolate (60% protection). In order to explain these vaccination results and to assess both the common and variable antigenicity of B. divergens, the antigenic patterns of the challenge isolates (Rouen 1987, 7107b, Weybridge 8843, and Munich 87) were compared by immunoprecipitation, using gerbil antisera raised against the Rouen 1987 vaccine isolate. Differences in the antigenic patterns and in the cross-protection of gerbils in these heterologous challenges were examined by studying the virulence and the antigenic status of each isolate.
Streptococcus pneumoniae produces at least two distinct enzymes with neuraminidase activity: cloning and expression of a second neuraminidase gene in Escherichia coli
Authors: Camara M , Mitchell T J , Andrew P W , Boulnois G J
Published: January 2020

A gene from Streptococcus pneumoniae was cloned in lambda EMBL301 and then expressed in Escherichia coli, which cleaved the fluorogenic neuraminidase substrate 2'-(4-methylumbelliferyl)-alpha-d-N-acetylneuraminic acid. The cloned gene therefore encodes an enzyme with neuraminidase activity. On the basis of restriction mapping and DNA hybridization studies, this gene could be distinguished from another pneumococcal neuraminidase gene cloned previously (A. M. Berry, J. C. Paton, E. M. Glare, D. Hansman, and D. E. A. Catcheside, Gene 71:299-305, 1988). Both neuraminidase genes were found in each of five isolates, covering at least three serotypes, of pneumococci tested.
Effects of activated macrophages on Mycobacterium leprae
Authors: Ramasesh N , Adams L B , Franzblau S G , Krahenbuhl J L
Published: January 2020

Five alternative methods were used to explore in vitro the effects of normal and activated murine macrophages on the metabolic well-being of intracellular Mycobacterium leprae: fluorescein diacetate-ethidium bromide staining, ATP content, synthesis of phenolic glycolipid 1, and two techniques to quantitate oxidation of palmitic acid. In relatively short-term experiments (7 to 10 days), each of these procedures provided strong evidence that activated macrophages exerted a deleterious effect on the leprosy bacillus. These findings appear to confirm the contention that activated macrophages underlie host resistance to clinical leprosy and limitation of M. leprae growth in paucibacillary leprosy.
Protective effect of Porphyromonas gingivalis outer membrane vesicles against bactericidal activity of human serum
Authors: Grenier D , Bélanger M
Published: January 2020

The present study was undertaken to evaluate the effect of Porphyromonas gingivalis outer membrane vesicles on the bactericidal activity of human serum. Human serum was pretreated with extracellular vesicles and then incubated with a cell suspension of Capnocytophaga ochracea. After 2 h at 37 degrees C, the percent viability of C. ochracea was determined by cultivation on blood agar plates. At a final concentration of 0.3 mg/ml, outer membrane vesicles completely inhibited the serum bactericidal activity against C. ochracea. Boiling the vesicles prevented this inhibition. However, partial inhibition of the serum lethal action was obtained when a higher concentration (1.5 mg/ml) of boiled vesicles was used, which indicates the involvement of both heat-labile and heat-stable components associated with vesicles. Combining vesicles at a suboptimal concentration (0.1 mg/ml) with a reducing agent brought back inhibition of the bactericidal activity, whereas combining vesicles at an optimal concentration (0.3 mg/ml) with a thiol-blocking reagent caused a restoration of the bactericidal activity. When a purified preparation of P. gingivalis lipopolysaccharides was used instead of vesicles, inhibition of the bactericidal activity was also observed. These results indicate that the lipopolysaccharides and the proteolytic enzyme(s) associated with P. gingivalis outer membrane vesicles are likely to represent the heat-stable and the heat-labile components, respectively. It is possible that outer membrane vesicles released by P. gingivalis protect other bacterial species from complement action, thus favoring the pathogenic process of periodontal disease.
The T-cell-stimulating 17-kilodalton protein of Francisella tularensis LVS is a lipoprotein
Authors: Sjöstedt A , Tärnvik A , Sandström G
Published: January 2020

A T-cell-stimulating, membrane-located 17-kDa protein of the live vaccine strain Francisella tularensis LVS has previously been cloned and sequenced. In the present study, it is shown to be a lipoprotein. When F. tularensis was grown in the presence of [3H]palmitate, several proteins of the organism, including a 17-kDa protein, were radiolabeled. The labeled 17-kDa protein was found by Western blot (immunoblot) analysis to be identical to the cloned protein. It was located in the detergent phase after partitioning with the nonionic detergent Triton X-114, thereby behaving like a hydrophobic integral membrane protein. The protein was predominantly hydrophilic and contained no putative transmembrane domain. The presence of fatty acids is therefore the probable explanation of the membrane location of the 17-kDa protein. The amino acid sequence of the 17-kDa protein contains the tetrapeptide Leu-Ala-Ser-Cys, which is a recognition sequence of the lipoprotein signal peptidase. Globomycin, a specific inhibitor of the peptidase, inhibited maturation of the 17-kDa lipoprotein. The protein incorporated [3H]palmitate also when expressed by Escherichia coli. The 17-kDa lipoprotein was recognized not only by T cells but also by serum antibodies of F. tularensis-primed individuals.
Activation of human monocytes by Streptococcus mutans serotype f polysaccharide: immunoglobulin G Fc receptor expression and tumor necrosis factor and interleukin-1 production
Authors: Benabdelmoumene S , Dumont S , Petit C , Poindron P , Wachsmann D , Klein J P
Published: January 2020

Streptococcus mutans serotype f polysaccharide (poly f) was prepared from S. mutans whole cells by autoclaving. The poly f was purified by chromatography on DEAE Trisacryl M and Bio-Gel P100, treated with insoluble pronase, and resubjected to chromatography on DEAE Trisacryl M. Normal human blood monocytes, stimulated in vitro with purified poly f, produced extracellular tumor necrosis factor alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) in a dose-dependent fashion as determined by a heterologous two-site sandwich enzyme-linked immunosorbent assay. Poly f also increased the expression of monocyte cell surface receptors for the Fc part of human immunoglobulin G, activity which is correlated with an increase of the phagocytic activity of the stimulated monocytes. Polymyxin B had no effect on TNF-alpha and IL-1 beta release. Neutralization assays with anti-recombinant human TNF-alpha and anti-recombinant human IL-1 beta immunoglobulin G confirmed the fact that the cytotoxic and mitogenic mediators released by the poly f-stimulated monocytes were mainly TNF-alpha and IL-1 beta.
Trypanosoma cruzi but not Trypanosoma brucei fails to induce a chemiluminescent signal in a macrophage hybridoma cell line
Authors: Vray B , De Baetselier P , Ouaissi A , Carlier Y
Published: January 2020

Macrophage-Trypanosoma cruzi interactions were studied by using a newly generated macrophage hybridoma cell line (2C11-12) that was selected for its capacity to produce high levels of reactive oxygen intermediates. This cell line was found to be a suitable host cell for T. cruzi, and intracellular parasitic development could be inhibited by activation with gamma interferon. When exposed to opsonized Trypanosoma brucei, Micrococcus lysodeikticus, or Legionella pneumophila, the activated macrophage cell line produces a high chemiluminescent signal, indicating the release of reactive oxygen intermediates. Alternatively, when opsonized T. cruzi was added to these activated macrophages, this parasite failed to stimulate a chemiluminescent response, suggesting an impairment in the triggering of the respiratory burst.
Sensitization of Mice by Inhalation of Antigen After Infection with
Bordetella pertussis
Authors: Chang Ik Chin , Gottshall R. Y.
Published: January 2020

Intranasal infection of mice with
Bordetella pertussis
or injection of pertussis vaccine previous to administration of an albumin aerosol augments sensitivity toward albumin. Sensitization was demonstrated by provocation of anaphylactic reactions following intravenous injection of antigen.
Migration Inhibitory Factor and Macrophage Bactericidal Function
Authors: Simon Harvey B. , Sheagren John N.
Published: January 2020

A homogeneous population of immunologically active lymphocytes was obtained from peritoneal exudates of guinea pigs with delayed hypersensitivity to bovine gamma globulin (BGG). The lymphocytes were cultured with and without BGG for 24 hr, and cell-free supernatant fluids were then assayed simultaneously for their ability to influence two in vitro parameters of macrophage function: migration from capillary tubes and bactericidal capacity. In four consecutive experiments, supernatants from antigenically stimulated lymphocytes exhibited substantial migration-inhibitory-factor activity without enhancing the ability of macrophages to kill
Listeria monocytogenes.
Lymphocyte lysates were inactive in both assays. Possible mechanisms of lymphocyte-macrophage interactions are discussed.
Location and Description of Spiral-Shaped Microorganisms in the Normal Rat Cecum
Authors: Davis Charles P. , Mulcahy D. , Takeuchi A. , Savage D. C.
Published: January 2020

Some indigenous microorganisms have been shown to localize in certain anatomical sites of the digestive tract of mammals. We studied the ceca of normal adult rats by light and electron microscopy to determine whether any specific bacterial population localizes in this area. All rats studied showed that the crypt was packed with organisms whose morphological character differs from those of the cecal lumen. Organisms localized in the crypt were often identified topographically close to the microvilli of the epithelial cells. These organisms could be differentiated into three types according to their characteristic ultrastructure. Type 1 was a thin spiral-shaped microbe that resembled a
Borrelia.
Type 2 possessed helically coiled fibers and flagella-like appendages. Type 3 was spiral-shaped but lacked axial fibers. Types 1 and 2 were both capable of penetrating through the crypt epithelium into the lamina propria where they were found in either phagocytes or extracellular locations. These observations are discussed in relation to other host-microflora localization patterns.
Polynucleotide Sequence Relatedness Among Three Groups of Pathogenic
Escherichia coli
Strains
Authors: Brenner Don J. , Fanning G. R. , Steigerwalt A. G. , Ørskov I. , Ørskov F.
Published: January 2020

Escherichia coli
strains that cause dysentery-like disease, parenteral infection, and infantile diarrhea form specific groups based on mobility of O and K antigens in immunoelectrophoresis. Members from each of these groups were assayed for gross nucleotide sequence relatedness. The method used was interspecific deoxyribonucleic acid (DNA) reassociation reactions carried out free in solution. Reassociated DNA was separated from unreacted DNA by passage through hydroxyapatite. DNA relatedness between these groups was approximately 80%. The groups containing those strains causing parenteral infection and those responsible for dysentery-like disease showed preferentially high intragroup DNA relatedness. The group containing strains responsible for infantile diarrhea did not show preferentially high intragroup DNA relatedness with the reference strain employed. These strains, however, did exhibit preferentially high DNA relatedness to a second reference strain.
Interferon Production Linked to Toxicity of Polyriboinosinic Acid-Polyribocytidylic Acid
Authors: de Clercq Erik , Stewart William E. , de Somer Pierre
Published: January 2020

Different procedures have been used in attempts to increase the production of interferon by polyriboinosinic acid-polyribocytidylic acid (poly [rI]·poly [rC]) in mice: simultaneous injection of lead acetate, cycloheximide, or actinomycin D and prior injection of Freund's adjuvant, chlorite-oxidized oxyamylose (COAM), endotoxin, or
Brucella abortus.
In the experimental conditions tested, lead acetate, cycloheximide, Freund's adjuvant, and COAM brought about a parallel increase in interferon production and toxicity (lethality) of poly (rI)·poly (rC); actinomycin D, endotoxin, and
B. abortus
increased the lethality of poly (rI)·poly (rC) without a concomitant raise of its interferon-inducing capacity. Our results indicate that no significant increase in interferon production (or antiviral activity, as far as the antiviral activity is accounted for by interferon production) without an accompanying increase in toxicity can be achieved with poly (rI)·poly (rC) and that it might be impossible to increase its therapeutic ratio (ratio of maximum tolerated dose to minimum effective dose).
Immunogenicity of the Common Enterobacterial Antigen Produced by Smooth and Rough Strains
Authors: Whang H. Y. , Mayer H. , Schmidt G. , Neter E.
Published: January 2020

Enterobacteriaceae
share a common antigen (CA). This antigen exists as a powerful immunogen, when produced by
Escherichia coli
O14, and as a minimally effective immunogen, when present in cultures of most other smooth strains. The present study was directed toward the assessment of immunogenicity of this antigen produced by various rough mutants and their parent strains as well as that of viable and nonviable bacterial suspensions. Rabbits were immunized intravenously, and the passive hemagglutination test was used for the quantitation of CA antibodies. The following results were obtained. (i) Immunization of rabbits with viable or heat-killed (1 hr, 100 C) suspensions of smooth parent strains resulted in only slight or insignificant antibody production. Of seven additional smooth strains of
Enterobacteriaceae
tested, none induced CA antibodies upon immunization with killed (1 hr, 100 C) suspensions, and only two engendered antibodies in moderate titers when viable suspensions were used, possibly due to the presence of undetected R mutant cells. (ii) In contrast, administration of viable suspensions of the corresponding mutant strains, regardless of core types, including
E. coli
R1, R2, R3, and
Salmonella
Ra types, caused a significant antibody response. (iii) The immunogenicity of R mutants differed strikingly, inasmuch as mutants with the R1 core [
E. coli
O8
−
:K27
−
(R) and
Shigella boydii
type 3
−
(R)] engendered CA antibodies when heated suspensions were administered, in contrast to heated suspensions of mutants with the R2, R3, and Ra cores. (iv) Viability per se is not an abolute requirement for immunogenicity of R2 mutants, since CA antibodies were engendered by suspensions killed at 60 C or by merthiolate or formaldehyde. (v) Ethanol fractionation revealed that immunogenic CA of the R1 mutant is ethanol-insoluble and that of the parental smooth strain is ethanol-soluble; thus, CA of R1 mutants resembles that of
E. coli
O14 in both immunogenicity and ethanol insolubility.
Role of Protein A in the Serum-Soft Agar Technique
Authors: Forsum Urban , Forsgren Arne , Hjelm Eva
Published: January 2020

Formation of compact colonies of
Staphylococcus aureus
in serum-soft agar is mainly a result of a reaction between protein A and the Fc-part of immunoglobulin G and not a clumping factor-fibrinogen reaction.
Factors Affecting Interaction of Staphylococcal Alpha Toxin with Membranes
Authors: Bernheimer Alan W. , Kim Kwang Shin , Remsen Charles C. , Antanavage Joanne , Watson Stanley W.
Published: January 2020

Staphylococcal alpha toxin interacts not only with membranes of erythrocytes but also with membranes of other kinds of mammalian cells (platelets, hepatocytes, and lysosomes from polymorphonuclear leukocytes) with the formation of characteristic ring-like structures that can be seen by electron microscopy. Such structures are not observed when alpha toxin is added to membranes derived from various bacteria. The rings seen on mammalian cell membranes tend to be either randomly disposed or in square array. The frequency with which square arrays are seen is influenced by the presence of staphylococcal delta toxin, by the negative staining agent, and by the kind of cell from which the membrane is derived. Synthetic membranes in the form of liposomes, prepared individually from phosphatidyl choline, phosphatidyl serine, phosphatidyl inositol, and cardiolipin, produced randomly disposed rings upon addition of alpha toxin. Liposomes made from phosphatidyl ethanolamine did not yield rings. Alpha toxin-treated liposomes prepared from chloroform-methanol extracts of brain white matter consistently showed rings that were rectangularly ordered. Ordered rings on membranes derived from toxin-treated platelets and those on toxin-treated brain extract liposomes were seen in freeze-etched as well as in negatively stained preparations.
Cell Surface Antigen Induced by Influenza Virus
Authors: Hahon Nicholas , Eckert Herbert L.
Published: January 2020

By immunofluorescence staining, a specific surface antigen induced by influenza virus was detected on clone 1-5C-4 cells. The antigen was neither related to hemagglutinin nor dependent on the production of infectious virus. Formation of the virus-specific cell surface antigen was dependent on protein synthesis but independent of viral ribonucleic acid replication.
Immunological and Chemical Studies of the Streptococcal Group O Protein Type Antigens
Authors: Mukasa H. , Slade H. D.
Published: January 2020

Two antigens present in group O streptococci have been described and are designated as type antigens. No other antigens are present in extracts of group O cells in significant quantity. Antigen I is a cell wall protein, and II is a nucleoprotein of undetermined cell location. The immunological specificity of the latter resides only in the protein. Both antigens have been extracted from the cell with water or 0.2
n
HCl (
p
H 2) at 100 C, and antigen I can be removed by trypsin. After purification,
p
H 2 was found to destroy the serological activity of each antigen. Antgens I and II and also the O polysaccharide antigen can be distinguished on a single agar-gel diffusion plate with group O whole-cell antiserum. N- and C-terminal amino acid analyses and total amino acid composition prove that I and II are separate molecular species. Each possesses properties which separate them from the streptococcal M, R, and T proteins. Antigen I separated into three fractions, each possessing serological specificity, on a diethylaminoethyl-Sephadex column at
p
H 8.6 during an increase in buffer molarity. The protein is considered to possess a multiple unit structure. The results indicate that the O streptococci are limited to two serological types. Group O whole-cell antisera, after adsorption with a strain not containing the O polysaccharide, can now be prepared for the positive identification of the types of group O streptococci.
Susceptibility of Rabbits Immunized with
Mycobacterium bovis
(BCG) or
Mycobacterium phlei
to
Shigella
Keratoconjunctivitis
Authors: Nakamura Mitsuru , Cross William R.
Published: January 2020

Vaccination of rabbits with mycobacteria increased their resistance to eye infection with
Shigella flexneri.
However, the severity of keratoconjunctivitis was not reduced in the vaccinated animals.
Detection of Staphylococcal Nuclease Elaborated During Lethal Infections in Mice
Authors: Chesbro William , Walker Richard
Published: January 2020

Active staphylococcal nuclease was detected in tissues of lethally infected mice. Organ distribution and levels of the enzyme varied with the bacterial strain used.
Leucocytosis-Promoting Factor of
Bordetella pertussis
I. Purification and Characterization
Authors: Sato Y. , Arai H.
Published: January 2020

The leucocytosis-promoting factor was purified from the supernatant fluid of spent cultures of
Bordetella pertussis
on solid medium. After precipitation at 67% saturation of ammonium sulfate, the leucocytosis-promoting factor was extracted with a 1.0
m
NaCl solution. Purification was accomplished by starch block electrophoresis and sucrose density gradient centrifugation. The purified preparation contained a high leucocytosis-promoting activity, and as small an amount as 0.04 μg of protein induced leucocytosis in mice. About 520-fold purification was attained, with a re-recovery of about 25% on an activity basis. The leucocytosis-promoting factor was composed solely of filamentous molecules of about 2 by 40 nm in size, with a sedimentation coefficient of approximately 5.5
S
and a molecular weight of 108,000. It was insoluble in water but partially soluble in 1.0
m
NaCl solution, and consisted mainly of protein, with some carbohydrate, lipid, and phosphorus.
Isolation and Purification of Endotoxin by Hydrolytic Enzymes
Authors: Lehrer Samuel , Nowotny Alois
Published: January 2020

Various commercial hydrolases were used in an attempt to degrade the endotoxic lipopolysaccharide macromolecule. Some inert components, such as peptides and nucleic acids, could be removed from endotoxin preparations. As a result, endotoxic activity, measured by pyrogenicity, Shwartzman reaction, and mouse lethality, was increased. The remarkable resistance of endotoxin to hydrolases led to the use of such enzymes for the liberation and purification of endotoxin from whole bacterial cells.
Infectious Antibody-Reovirus Complexes
Authors: Huggett David O. , Rodríguez José E. , McKee Albert P.
Published: January 2020

Populations of reovirus type 1 could not be completely neutralized by exposure to specific antiviral serum. Aggregation of reovirions before or after exposure to neutralizing antibody was discarded as a possible explanation for the phenomenon observed. Anti-rabbit serum and equilibrium zonal density gradient centrifugation were used to show that the residual infectious fraction consisted of infectious antibody-virion complexes. Electron microscopy and equilibrium density gradient centrifugation of stock virus preparations were used to attempt to determine whether the infectious virus-antibody complexes resulted from the combination of antibody with virus particles which differed from neutralizable virions either morphologically or with regard to buoyant density. Although electron micrographs revealed the presence of three morphologically distinct classes of virus particles, density gradient centrifugation resulted in only one infectious peak. Density gradient centrifugation of virus exposed to antibody resulted in only one infectious peak which consisted of virus-antibody complexes.
Experimental congenital syphilis: guinea pig model
Authors: Wicher K , Baughn R E , Wicher V , Nakeeb S
Published: January 2020

Neonates born to female guinea pigs of either a highly susceptible (C4D) or a resistant (Albany) strain, infected prior to or during pregnancy with a single dose of Treponema pallidum, showed in their sera from the first day of life immunoglobulin M (IgM) antibodies to T. pallidum, circulating immune complexes consisting of IgM antibodies and treponemal antigens, and IgM rheumatoid factor. Although the animals were asymptomatic for a 6-month observation period, several lines of evidence indicated that they were infected in utero. Molecular analysis of whole sera, purified serum IgM fraction, or dissociated immune complexes demonstrated IgM reactivity against one (47 kDa) or more of several T. pallidum peptides (15, 17, 37, 42, 45, and 87 kDa) recognized as integral membrane components. Sequential analysis of the neonates' sera by immunoblot and enzyme-linked immunosorbent assay, using alcohol-treated T. pallidum, T. phagedenis biotype Reiter, and T. vincentii, demonstrated early IgM antibodies followed 3 to 4 months later by IgG2- and IgG1-specific antibodies to T. pallidum. Moreover, an infectivity test done in five rabbits with pooled tissue extracts prepared from liveborn or stillborn animals evoked a seroconversion in two rabbits (reactive Venereal Disease Research Laboratory and fluorescent treponemal antibody tests), suggesting the presence of T. pallidum in the organs. Sera from neonates born to either T. phagedenis biotype Reiter-injected mothers or three normal pregnant females were all serologically negative. The model offers new possibilities for exploration of factors responsible for asymptomatic infection often observed in human congenital syphilis.
Differential effects of monoclonal antibodies to tumor necrosis factor alpha and gamma interferon on induction of hepatic nitric oxide synthase in experimental gram-negative sepsis
Authors: Evans T , Carpenter A , Silva A , Cohen J
Published: January 2020

To investigate the stimuli required for the induction of nitric oxide synthase (NOS) in sepsis, we have analyzed the levels of this enzyme in the livers of mice infected with a 90% lethal dose of Escherichia coli in a model of gram-negative sepsis. Hepatic NOS levels are markedly induced in this model, with peak values occurring 12 to 22 h following infection. Treatment with TN3-19.12, a neutralizing monoclonal antibody to tumor necrosis factor alpha (TNF-alpha), resulted in complete protection from death in this model of sepsis but had no significant effect on the level of induction of hepatic NOS. Treatment with H22, a monoclonal antibody to gamma interferon (IFN-gamma), also gave significant protection against death and, in addition, did lead to a decrease in the level of induction of the hepatic NOS. Treatment of mice with pure TNF-alpha (0.2 microgram), IFN-gamma (2,000 U), or a combination of the two did not induce the hepatic NOS, but treatment with the combination led to significant mortality (probability of survival at 22 h, 0.32). Thus, the level of induction of NOS within the liver either in sepsis or by the coadministration of TNF-alpha and IFN-gamma does not correlate with death.
Induction of procoagulant activity on human endothelial cells by Streptococcus pneumoniae
Authors: Geelen S , Bhattacharyya C , Tuomanen E
Published: January 2020

The inflammatory response in infection caused by gram-negative organisms involves induction of procoagulant activity (PCA) on human endothelial cells. Although infections caused by gram-positive organisms are also associated with fibrin formation and thrombosis, the bacterial determinants inducing PCA are unknown. This study shows that intact pneumococci and the pneumococcal cell wall efficiently induce PCA on human endothelial cells. Upon exposure of endothelial cells to pneumococci, PCA was first detectable at 30 min, peaked at 2 h, and disappeared by 6 h. The specific activities of encapsulated and unencapsulated strains for induction of PCA were equivalent. Purified pneumococcal cell walls were as potent as endotoxin in induction of PCA. The ability to induce a procoagulant state on endothelial cells is a new biological activity of gram-positive cell walls which promotes the participation of the coagulation cascade in the inflammatory response to disease caused by gram-positive organisms.
Specific lung mucosal and systemic immune responses after oral immunization of mice with Salmonella typhimurium aroA, Salmonella typhi Ty21a, and invasive Escherichia coli expressing recombinant pertussis toxin S1 subunit
Authors: Walker M J , Rohde M , Timmis K N , Guzmán C A
Published: January 2020

Pertussis toxin (PT) is considered an essential protective component for incorporation into new generation vaccines against Bordetella pertussis, the causative agent of whooping cough. Traditionally, antipertussis vaccination has employed an intramuscular route. An alternative to this approach is to stimulate mucosal and systemic immune responses by oral immunization with live vaccine carrier strains of Salmonella spp. or Escherichia coli. Recombinant S1 subunit of pertussis toxin was expressed in the attenuated aroA mutant of Salmonella typhimurium, SL3261, in the human typhoid vaccine strain Salmonella typhi Ty21a, and in E. coli CAG629 containing the Shigella flexneri plasmid pWR110, which encodes bacterial invasiveness of epithelial cells. Expression of recombinant PT S1 subunit (rPT-S1) did not affect in vitro invasiveness of the tested strains, which retained the ability to adhere to and invade the embryonic human intestinal cell line HI-407. Following oral immunization of mice with the live vaccine strains expressing rPT-S1, immunoglobulin G (IgG), IgA, and IgM responses were monitored. IgG specific to PT was detected in serum samples of mice, while IgG and IgA specific to PT were detected in lung washes after oral immunization with living Salmonella spp. or E. coli (pWR110) expressing rPT-S1. Utilization of live oral vaccines expressing B. pertussis antigens, which stimulate both a systemic and lung mucosal response, may provide an attractive alternative to purified component vaccines against whooping cough.
Identification of 2,3-dihydroxybenzoic acid as a Brucella abortus siderophore
Authors: López-Goñi I , Moriyón I , Neilands J B
Published: January 2020

Brucella abortus grown in low-iron medium or in the presence of iron chelators [ethylenediamine-di(o-hydroxyphenylacetic acid) and 2,2-dipyridyl] showed reduced cell yields and released a material positive in chemical and biological assays for catechols. This material was purified from culture fluids of B. abortus 2308 by chromatography on agarose-iminodiacetic acid-Fe3+ and identified as 2,3-dihydroxybenzoic acid (2,3-DHBA) by thin-layer chromatography, paper electrophoresis, and UV-visible nuclear magnetic resonance and mass spectroscopy. No other major catechols were observed at different stages of growth, and 2,3-DHBA was also produced upon iron limitation by representative strains of B. abortus biotypes 1, 5, 6, and 9. Both synthetic 2,3-DHBA and the natural catechol relieved the growth inhibition of B. abortus 2308 by ethylenediamine-di(o-hydroxyphenylacetic acid), and 2,3-DHBA promoted 55Fe uptake by B. abortus 2308 by an energy-dependent mechanism. Two other monocatechols tested, 2,3-dihydroxybenzoyl-Ser and 2,3-dihydroxybenzoyl-Gly, also promoted 55Fe uptake. More complex catechol siderophores (agrobactin and enterobactin), hydroxamate siderophores (aerobactin, ferrichrome, and deferriferrioxamine mesylate [Desferal]), and an EDTA-related siderophore (rhizobactin) failed to mediate 55Fe uptake. B. abortus cells grown in low-iron medium or in medium with iron had similar rates of iron uptake when supplied with 55Fe-2,3-DHBA, and the release of 2,3-DHBA under iron starvation was not associated with the expression of new outer membrane proteins. These results suggest an uptake system in which only the synthesis of the siderophore is regulated by the iron available for growth.
Rapid generation of specific protective immunity to Francisella tularensis
Authors: Elkins K L , Leiby D A , Winegar R K , Nacy C A , Fortier A H
Published: January 2020

Mice inoculated either subcutaneously (s.c.) or intradermally (i.d.) with a sublethal dose of Francisella tularensis LVS are immune to a lethal intraperitoneal (i.p.) or intravenous (i.v.) challenge of LVS. Here, we show that this immunity developed quite rapidly: mice given a sublethal dose of live LVS s.c. or i.d. (but not i.v.) withstood lethal i.p., i.v., or i.d. challenge as early as 2 days after the initial inoculation, despite the presence of bacterial burdens already in tissues. The magnitude of this early protection was quite impressive. The i.p. 50% lethal dose (LD50) in naive C3H/HeN mice was only 2 bacteria, while the i.p. LD50 in mice given 10(4) LVS i.d. 3 days previously was 3 x 10(6) bacteria. Similarly, the i.v. LD50 in C3H/HeN mice shifted from 3 x 10(2) in naive mice to 5 x 10(6) in primed mice within 3 days after i.d. LVS infection. Comparable changes in the i.p. and i.v. LD50 were observed in C57BL/6J mice. This rapid generation of protective immunity was specific for LVS, in that mice given a sublethal i.d. inoculation of LVS did not survive a lethal challenge with either Salmonella typhimurium W118 or Escherichia coli O118 BORT at any time, nor could mice given sublethal doses of S. typhimurium, E. coli, or Mycobacterium bovis BCG survive lethal doses of LVS. Although an increase in the mean time to death from S. typhimurium infection was noted when mice were given a sublethal i.d. dose of LVS 4 to 14 days earlier, no overall increase in protection or change in the S. typhimurium LD50 was observed. Thus, sublethal infection with LVS at skin sites induced rapid and specific protective immunity.
Colony-stimulating factors activate human macrophages to inhibit intracellular growth of Histoplasma capsulatum yeasts
Authors: Newman S L , Gootee L
Published: January 2020

Recombinant cytokines and colony-stimulating factors (CSFs) were tested for their abilities to activate human monocytes/macrophages (M phi) to inhibit the intracellular growth of or kill Histoplasma capsulatum yeasts. None of the cytokines or CSFs or combinations of cytokines and CSFs activated M phi fungistatic activity when they were added to M phi monolayers concurrently with yeasts. In contrast, culture of monocytes for 7 days in the presence of interleukin 3, granulocyte-M phi CSF, or M phi CSF stimulated M phi fungistatic (but not fungicidal) activity against H. capsulatum yeasts in a concentration-dependent manner. Optimal activation of M phi by CSFs required 5 days of coculture, and the cultures had to be initiated with freshly isolated peripheral blood monocytes. Culture of monocytes with combinations of CSFs or addition of CSFs during the 24 h of coculture with the yeasts did not further enhance M phi fungistatic activity for H. capsulatum. Addition of gamma interferon or tumor necrosis factor alpha to CSF-activated M phi also did not enhance M phi fungistatic activity. These results suggest that interleukin 3, granulocyte-M phi CSF, and M phi CSF may play a role in the cell-mediated immune response to H. capsulatum by enhancing monocyte/M phi fungistatic activity.
Major outer membrane proteins of Vibrio cholerae and their role in induction of protective immunity through inhibition of intestinal colonization
Authors: Sengupta D K , Sengupta T K , Ghose A C
Published: January 2020

Vibrio cholerae O1 organisms belonging to different biotypes and serotypes were shown to express major outer membrane proteins (MOMPs) with subunit molecular masses of 48 to 50, 40 to 43, 35 to 36, 27 to 28, and 20 kDa. Antisera raised against individual MOMPs of a V. cholerae O1 strain recognized MOMPs of corresponding molecular masses in other O1 and non-O1 strains. Serological data also suggested possible differences in the cell surface exposition of these MOMPs. However, no marked differences between V. cholerae cells grown in vitro and in vivo could be noted in respect to the expression or surface exposition of these MOMPs. Of five MOMPs studied in this work, 40- to 43- and 20-kDa cell surface proteins were shown to be of considerable importance, as antisera to these proteins induced significant protection against V. cholerae challenge in the suckling mouse model. Similar protection, although to a lesser extent, was demonstrable with the antiserum to the 27- to 28-kDa protein. These results were corroborated with the Fab (immunoglobulin G) [Fab(IgG)] fragments of the antisera, thereby suggesting that the observed protection induced by anti-MOMP antibodies did not arise as a result of bacterial clumping. Subsequent studies demonstrated that these antisera as well as their Fab (IgG) fragments induced significant inhibition of intestinal colonization of V. cholerae. The 40- to 43- and 27- to 28-kDa proteins appeared to be porinlike, while the 20-kDa protein was found to be antigenically related to TcpA (subunit A of toxin-coregulated pilus). All these results demonstrate the involvement of more than one cell surface antigen of V. cholerae in the induction of protective immunity through inhibition of intestinal colonization of vibrios.
Characterization and protective properties of attenuated mutants of Salmonella choleraesuis
Authors: Kelly S M , Bosecker B A , Curtiss R
Published: January 2020

We have constructed crp::Tn10 and cya::Tn10 Salmonella choleraesuis mutants and their fusaric acid-resistant derivatives with deletions (delta) of the Tn10 and adjacent DNA sequences and found them to be avirulent and able to induce protection against a wild-type challenge in 8-week-old BALB/c mice. Mice survived infection with the crp and cya mutants at doses of more than 7 x 10(3) times the oral (p.o.) 50% lethal dose (LD50) and more than 8 x 10(2) times the intraperitoneal LD50 of the wild-type S. choleraesuis parent. Mice vaccinated with attenuated strains were protected against challenge with more than 1.6 x 10(4) times the p.o. LD50 and more than 80 times the intraperitoneal LD50 of the wild-type virulent S. choleraesuis parent. One deletion mutation isolated in the crp region extends to an adjacent gene(s) that was shown to be associated with avirulence. This gene or operon has been designated cdt (colonization of deep tissues). A delta (crp-cdt)19 strain, when complemented with the wild-type crp gene and promoter on a pBR-derived plasmid, had p.o. LD50 values 10(3) times higher than those for the wild type. A delta cya delta (crp-cdt)19 double mutant was less virulent than and afforded more complete protection against a challenge with the wild-type strain than a delta crp-11 delta cya double mutant or the individual cya, crp, or crp+/cdt mutants. The deletion derivatives exhibited reduced invasion of CHO cells in vitro, and the numbers of the mutants recovered from mouse tissues were less than that of the parent strain. These studies suggest that one or more of the genes involved in cell attachment to and/or invasion of S. choleraesuis may be under catabolite repression. In addition, we describe a new deletion of a gene(s) located in the crp region between cysG and argD that is associated with virulence in S. choleraesuis.