bluuebunny/crossref_metadata_2025 huggingface dataset:
An outbreak of gastroenteritis due to a heat-labile enterotoxin-producing strain of Escherichia coli
Authors: Wood L V , Wolfe W H , Ruiz-Palacios G , Foshee W S , Corman L I , McCleskey F , Wright J A , DuPont H L
Published: January 2020

In April 1981, an outbreak of gastroenteritis, characterized by diarrhea and abdominal cramps, occurred in 282 of approximately 3,000 personnel at a large metropolitan hospital in San Antonio, Tex. There was a significant association between illness and eating at the hospital cafeteria (P = 0.0008), but no specific food could be incriminated. Stools or rectal swabs from 54 ill individuals produced almost pure cultures of Escherichia coli. Cultures from 51 of these subjects had identical antibiotic sensitivity patterns, and 38 had the same biotype. Isolates from 45 persons were tested for production of heat-stable and heat-labile enterotoxins, using the suckling mouse and Y-1 adrenal cell assays, respectively. Of 45 isolates, 41 produced heat-labile enterotoxins, while 0 of 45 produced heat-stabile enterotoxins. Two isolates were rough, and 34 of the remaining 43 were serotype O25:H-. Two strains were O25:H+. None of the 45 strains possessed hemagglutination patterns typical of colonization factor antigens I or II. Six of seven O25:H- heat-labile enterotoxin-positive strains selected at random were piliated as seen by electron microscopy but did not agglutinate with anti-colonization factor antigens I or II antisera.
Characterization of binding of human alpha 2-macroglobulin to group G streptococci
Authors: Chhatwal G S , Müller H P , Blobel H
Published: January 2020

An interaction was observed between human alpha 2-macroglobulin (alpha 2M) and streptococci belonging to group A, C, and G. Of 27 group C and 19 group G streptococcal cultures, 13 and 14, respectively, bound 125I-labeled alpha 2M. Some group A streptococci also interacted with alpha 2M. A number of other bacterial species tested did not react with alpha 2M. The binding of 125I-labeled alpha 2M to group G streptococci was time dependent, saturable, and could be inhibited by unlabeled alpha 2M. Inhibition experiments indicated that the streptococcal binding site for alpha 2M differed from the receptors for immunoglobulin G, fibrinogen, aggregated beta 2-microglobulin, albumin, and fibronectin. The alpha 2M binding activity was remarkably sensitive to trypsin and heat treatment indicating its protein nature. Kinetic analysis indicated a homogenous population of binding sites. The number of binding sites per bacterial cell was estimated to be approximately 20,000.
Isolation and properties of levanase from Streptococcus salivarius KTA-19
Authors: Takahashi N , Mizuno F , Takamori K
Published: January 2020

Fructan-hydrolyzing enzyme from Streptococcus salivarius KTA-19 isolated from human dental plaque was investigated. The enzyme was purified by ammonium sulfate precipitation, acetone fractionation, and column chromatography on Bio-Gel and DEAE-cellulose. The purified enzyme showed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing. Its molecular weight was 100,000 as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme exhibited an optimum pH of 6.5 and decreased its activity from pH 6.0 and especially below pH 5.5. The optimum temperature was 40 to 50 degrees C, and enzyme activity was reduced by 90% at 55 degrees C. Enzyme activity was markedly inhibited by Hg2+, Ag+, Cu2+, and p-chloromercuribenzoate at a concentration of 10(-3) M, but not by other metal ions or chemical effectors. Fructose was the only by-product of the enzyme action on levan. These results indicated that the levanase of S. salivarius KTA-19 is an exo-beta-(2,6)-fructofuranosidase.
Common lipopolysaccharide specificity: new type of antigen residing in the inner core region of S- and R-form lipopolysaccharides from different families of gram-negative bacteria
Authors: Brade H , Galanos C
Published: January 2020

A new antigenic specificity, referred to here as common lipopolysaccharide (LPS) specificity, is described in the LPSs of gram-negative bacteria belonging to various families. The specificity is present in S- and R-form LPS but absent in Re mutants of different enterobacterial genera. By the use of purified LPS and monospecific antibodies obtained by immunoabsorption, the specificity is differentiated from the known core specificities of the genus Salmonella and the lipid A specificity by aid of the passive hemolysis and passive hemolysis inhibition test. In Salmonella minnesota R-form LPS, the specificity may be cryptic (R345, Rb2 mutant) or partly exposed in the intact molecule (R7, Rd1 mutant). The specificity is either demasked or completely exposed after mild acid hydrolysis for a short time, whereas it is destroyed after prolonged hydrolysis. Periodate oxidation, reduction, and hydrolysis under conditions that do not affect the ketosidic linkages of 2-keto-3-deoxyoctulosonic acid destroy the specificity in R4 (Rd2 mutant) LPS, but do not do so in R7 LPS. It is suggested that 2-keto-3-deoxyoctulosonic acid and a following neutral sugar are the compositional requirements for expressing the specificity.
Effect of immune serum on infectivity of Rickettsia tsutsugamushi
Authors: Hanson B A
Published: January 2020

Hyperimmune antirickettsial serum was shown to prevent the attachment/penetration stage of Rickettsia tsutsugamushi infection of suspended chicken cells. The extent of the inhibition depended on the serum concentration but not on the presence of complement. The neutralizing activity was reduced by prior adsorption of immune serum with staphylococcal protein A or with intact rickettsiae but was not affected by adsorption with target cells. In the neutralization tests, there was no cross-reactivity between the Karp and Gilliam strains of R. tsutsugamushi. Incubation of rickettsiae with immune serum did not alter their capacity to metabolize glutamate nor grossly damage the permeability barrier function of their cytoplasmic membranes. Although the assay method had the capacity to detect some aggregated infectious organisms, none were found in immune serum-treated suspensions. It was concluded that immune serum may inhibit rickettsial infection by blocking a surface component(s) whose function is necessary for attachment to and/or penetration of target cells.
In vivo microscopic observations of the responses of Kupffer cells and the hepatic microcirculation to Mycobacterium bovis BCG alone and in combination with endotoxin
Authors: McCuskey R S , Urbaschek R , McCuskey P A , Urbaschek B
Published: January 2020

Kupffer cell function and hepatic microvascular hemodynamics were studied by in vivo microscopy in Mycobacterium bovis BCG-infected NMRI mice before and after treatment with minute (0.01 mu mg) tolerance-producing doses and doses causing 70% lethality (0.5 micrograms) of Escherichia coli 0111:B5 endotoxin alone and in combination. BCG-induced granulomas distorted the hepatic microvasculature and impeded blood flow in many sinusoids; flow also was altered further by leukocytes adhering to the sinusoidal walls and by enlarged Kupffer cells that bulged into the lumen. Nevertheless, in BCG-infected mice, the ratio of Kupffer cells which phagocytosed latex to sinusoids containing blood flow and capable of delivering these particulates to Kupffer cells was significantly greater than that in uninfected mice. The phagocytosis of single latex particles by individual Kupffer cells also was more rapid. This indicated an expansion of the numbers and activation of Kupffer cells. In this hyperreactive state, the tolerance-inducing dose of endotoxin produced no change in the rate of phagocytosis after 2 h. In contrast, the 70% lethal dose reduced the rate by 123%, unless tolerance was induced, in which case there was no reduction in the rate of phagocytosis. Twenty-four hours after injection of the tolerance-inducing dose, however, the rate of phagocytosis was accelerated slightly (17%). This suggested that the Kupffer cells had been activated and perhaps were more effective in clearing subsequent endotoxin from the blood but without sufficient release of toxic substances to be lethal. That some mediators were released, however, was suggested by the microvascular alterations that accompanied the above phagocytic responses. These results further support the concept of a central role for Kupffer cells in endotoxin-mediated, nonspecific host defense mechanisms.
Mechanisms of immunity to respiratory syncytial virus in cotton rats
Authors: Prince G A , Horswood R L , Camargo E , Koenig D , Chanock R M
Published: January 2020

Active immunity and maternally transmitted passive immunity to respiratory syncytial virus (RSV) were studied in cotton rats. Animals infected with respiratory syncytial virus developed complete resistance to pulmonary reinfection, which lasted at least 18 months. Nasal resistance was of shorter duration and began to diminish in 8 months. Pulmonary resistance was transferred by parabiosis, but nasal resistance was not. Adoptive transfer studies with fractionated convalescent blood showed that serum antibody, but not circulating lymphocytes, conferred pulmonary resistance. Immune females conferred antibody to their young prenatally and postnatally, with most of the antibody being transferred via colostrum and milk. Maternally transmitted immunity was more effective in the lungs than in the nose and was transient in both organs. Foster nursing experiments showed colostrum and milk to be the most important routes of immune transfer. Although resistance in infants generally correlated with serum neutralizing antibody levels, several exceptions to this correlation suggested that immune factors other than neutralizing antibody may also play an important role in maternal passive immunity.
Phagocytic killing of Neisseria gonorrhoeae by human monocytes
Authors: Mezzatesta J R , Rest R F
Published: January 2020

The ability of human monocytes to phagocytize and kill nonpiliated opaque (T3) and transparent (T4) gonococci was investigated in a tumbling tube suspension assay. A serum-sensitive strain, F62, and a serum-resistant strain, FA19, were studied. CFU remaining after incubation with monocytes were used to assess the extent of killing. The data show that 50% of T3 and T4 gonococci of both strains were killed by monocytes over a 2-h period. Serum was necessary for the killing of transparent gonococci of both strains as well as for FA19 T3. Concentrations of serum ranging from 0.5 to 10% were equally effective, and heat-labile components were required. Killing of F62 T3, however, occurred in the absence of serum. An increased ratio of bacteria to monocytes decreased the rate of killing. A 30-min preopsonization of gonococci in 10% serum resulted in an enhanced rate of killing. Monocytes were able to kill plate-grown, but not log-phase, organisms. Disruption of the monocytes by sonication to release internalized bacteria did not increase the number of viable organisms. The addition of 10 micrograms of cytochalasin B per ml completely inhibited the reduction in colony numbers over time. These data indicate that freshly isolated human monocytes are capable of phagocytizing and killing nonpiliated gonococci.
A wild and an attenuated strain of Francisella tularensis differ in susceptibility to hypochlorous acid: a possible explanation of their different handling by polymorphonuclear leukocytes
Authors: Löfgren S , Tärnvik A , Thore M , Carlsson J
Published: January 2020

We have previously reported that a wild strain of Francisella tularensis is much less efficiently killed by human polymorphonuclear leukocytes than is an attenuated strain. In the present study, the killing of the attenuated strain was found to be strictly oxygen dependent. The wild and the attenuated strains both induced a respiratory burst in the leukocytes. The difference between the strains in susceptibility to agents produced at the burst could be explained by a difference in susceptibility to hypochlorous acid.
Killing of Aspergillus fumigatus spores and Candida albicans yeast phase by the iron-hydrogen peroxide-iodide cytotoxic system: comparison with the myeloperoxidase-hydrogen peroxide-halide system
Authors: Levitz S M , Diamond R D
Published: January 2020

A new fungicidal system composed of ferrous ion, H2O2, and iodide is described and compared with the myeloperoxidase-hydrogen peroxide-halide system. Both systems had similar activity against Aspergillus fumigatus spores and the Candida albicans yeast phase, but only the ferrous ion-hydrogen peroxide-iodide system was inhibited by hydroxyl radical scavengers.
Bacterial adherence and glycocalyx formation in osteomyelitis experimentally induced with Staphylococcus aureus
Authors: Mayberry-Carson K J , Tober-Meyer B , Smith J K , Lambe D W , Costerton J W
Published: January 2020

A surgical procedure allowed the placement of a silicone rubber catheter in the marrow cavity of the tibia of a rabbit and also allowed the introduction of a sclerosing agent (sodium morrhuate) and cells of Staphylococcus aureus. Osteomyelitis developed in 60% of the animals so treated, and the infecting microorganism was recovered from the infected tibias of the animals that developed this disease. All blood cultures taken 24 h after the infection were negative for S. aureus. Radiological findings consisted of osteolytic changes, the occurrence of sequestration and periosteal reactions, and sclerosis in the infected bones. Sections of bone prepared for histological examination confirmed the diagnosis of osteomyelitis. Transmission and scanning electron microscopy of samples of bone marrow, bone chips, and the catheters taken from the infected tibiae revealed gram-positive cocci embedded in a very extensive matrix of ruthenium red-staining glycocalyx adhering to the bone and the implanted catheter. It is proposed that this extensive glycocalyx served a protective function for the bacteria and was important in bacterial adherence and thus played an important role in bacterial persistence and the development of osteomyelitis in these rabbits.
Trypanosoma musculi infection in B-cell-deficient mice
Authors: Vargas F D , Viens P , Kongshavn P A
Published: January 2020

The course of infection with Trypanosoma musculi was assessed in mice deprived of B-lymphocytes by administration, from birth, of rabbit antiserum to mouse immunoglobulin M (IgM). Initial control of parasitemia leading to the first crisis and establishment of the plateau phase was unaffected by lack of B-lymphocyte function, although multiplicative forms persisted throughout the infection in anti-IgM-treated mice, instead of disappearing after the first crisis as in intact mice. Elimination of trypanosomes after the second crisis was not observed in anti-IgM-treated mice, which maintained high numbers of parasites in the blood and peritoneal cavity, resulting in some mortality. A temporary reduction in parasitemia was achieved in anti-IgM-treated mice by transfusion of immune plasma. Immunodepression, as measured by splenic mitogen responsiveness, and splenomegaly were both observed in anti-IgM-treated as well as in intact mice, indicating that these features of murine trypanosomiasis are independent of B-lymphocyte function. Since in T. musculi infection parasitemia can be controlled initially but not eliminated in mice lacking B-cell function, the only crucial protection provided by antibody would appear to be in curing the infection after the second crisis.
Generation of leukotrienes from human granulocytes by alveolysin from Bacillus alvei
Authors: Bremm K D , Brom H J , Alouf J E , König W , Spur B , Crea A , Peters W
Published: January 2020

We investigated the effect of alveolysin on human granulocytes. Alveolysin is an exoprotein produced by Bacillus alvei and belongs to the group of sulfhydryl-activated cytolysins. Other members of this group are streptolysin O and theta-toxin from Clostridium perfringens. It is demonstrated that alveolysin leads to leukotriene generation from human granulocytes, which exert chemotactic (leukotriene B4) and slow-reacting substance (leukotriene C4, D4, and E4) activity under sublytic concentrations.
Degradation of human immunoglobulins by proteases from Streptococcus pneumoniae obtained from various human sources
Authors: Wikström M B , Dahlén G , Kaijser B , Nygren H
Published: January 2020

The ability of Streptococcus pneumoniae to degrade human secretory immunoglobulin A (S-IgA), IgG, and IgM was tested in 102 strains by use of the thin-layer enzyme assay cultivation technique. The strains were isolated from patients with acute phases of otitis media, meningitis, and pneumonia as well as from symptomless carriers. An ability to degrade S-IgA, IgG, and IgM was revealed in 50, 84, and 96 strains, respectively. An IgG- and IgM-degrading ability of S. pneumoniae has not previously been reported. A concurrent degradation of the three immunoglobulins was revealed in 38 strains; degradation of two of them was revealed in 54 strains, and degradation of only one of them was revealed in 9 strains. One strain failed to degrade any of the immunoglobulins. Correlations were not found between the ability of the S. pneumoniae strains to degrade S-IgA, IgG, or IgM and the serotype affiliation or between the ability to degrade IgG or IgM and the origin of strains. However, the ability to degrade S-IgA was evident more often in strains isolated from symptomless carriers and from bronchial secretions of patients with acute pneumonia than it was in strains from patients with acute meningitis or acute otitis media or from the blood of patients with acute pneumonia. These latter findings may indicate a biological significance of S-IgA-degrading ability in bacterial colonization of mucosal surfaces.
Oxygen radical production by peritoneal macrophages and Kupffer cells elicited with Lactobacillus casei
Authors: Hashimoto S , Nomoto K , Matsuzaki T , Yokokura T , Mutai M
Published: January 2020

BALB/c mice were injected intraperitoneally (i.p.) or intravenously (i.v.) with Lactobacillus casei YIT9018 (LC 9018). The i.p. injected LC 9018 augmented oxygen radical (OR) production by peritoneal macrophages (PM) and suppressed the production of prostaglandin E2 by PM. The growth of i.p. inoculated Meth A fibrosarcoma was also inhibited by an i.p. injection of LC 9018. i.v. injection of LC 9018 stimulated OR production by fixed macrophages and inhibited the growth of Listeria monocytogenes in the liver. Furthermore, glucose-6-phosphate dehydrogenase activity in the liver was significantly increased (10 to 20 times) by LC 9018 i.v. injection. A significant correlation was observed between the augmentation of OR production by PM or fixed macrophages in the liver and inhibition of growth of Meth A or L. monocytogenes. The augmentation of OR production by LC 9018 was more marked and was maintained for a longer period of time than that by other bacterial immunostimulants.
Cytolytic activity and virulence of Vibrio damsela
Authors: Kreger A S
Published: January 2020

A correlation was observed between the ability of 19 isolates of Vibrio damsela, a halophilic bacterium recently recognized as a human pathogen, to cause disease in mice and to produce large amounts of a cytolytic toxin in vitro. The yield of toxin in the culture supernatant fluids was optimal during the mid- and late-logarithmic phases of growth in medium containing 0.5% Na+ ion, was stable during the stationary growth phase, and was significantly reduced in culture medium containing greater than or equal to 0.8% Na+ ion, even though Na+ ion concentrations ranging from 0.8% to 2% significantly enhanced growth of the bacterium. The activity in toxin preparations partially purified by ammonium sulfate precipitation was deleteriously affected by heat, low and high pH, proteases, dithiothreitol, and chelating agents, but was unaffected by cholesterol, trypan blue, and mixed gangliosides. The toxin had a molecular weight (estimated by gel filtration) of ca. 57,000 and an isoelectric point of ca. 5.7 and was antigenically distinct from previously described cytolysins produced by Vibrio vulnificus, Vibrio parahaemolyticus, and the El Tor biotype of Vibrio cholerae. Bacteriologically sterile, partially purified toxin preparations were lethal for mice after intraperitoneal, intravenous, and subcutaneous administration, and subcutaneous injection elicited grossly observable changes similar to those observed during the lethal experimental infection caused by subcutaneous injection of V. damsela.
Mitogenic activities of synthetic lipid A analogs and suppression of mitogenicity of lipid A
Authors: Tanamoto K , Galanos C , Lüderitz O , Kusumoto S , Shiba T
Published: January 2020

The effect of synthetic lipid A analogs on murine spleen cells was studied. The preparations represented D-glucosamine and D-glucosaminyl-beta 1,6-D-glucosamine disaccharide derivatives substituted in different combinations by ester- and amide-bound fatty acids and by phosphate groups. Significant mitogenic activity was demonstrated with a number of synthetic disaccharide preparations; however, their potency was lower than that of lipid A. The synthetic preparations were not mitogenic for spleen cells from C3H/HeJ mice. Furthermore, the mitogenicity of the synthetic preparations was abolished after binding with polymyxin B. The results indicate that for expression of mitogenicity, a phosphate group at position 1 of the reducing glucosamine and amide-bound acyloxyacyl residues are important factors. Some of the synthetic preparations containing the diglucosamine backbone and expressing relatively low mitogenicity suppressed B-cell mitogenicity of lipid A. Although these preparations were lytic for erythrocytes, they did not affect the viability of the splenic lymphocytes. Suppression was seen when the synthetic preparations were added simultaneously with or after the lipid A mitogen, but optimal suppression was expressed when the preparations were added to the system 3 h before lipid A. Washing of the cells before the addition of lipid A did not affect the results. The suppression was not due to the induction of suppressor cells by the synthetic preparations. The disaccharide preparations did not inhibit T-cell mitogenicity of concanavalin A. In contrast to the disaccharide preparations, the monosaccharide preparations suppressed mitogenicity of both lipid A and concanavalin A, probably because of their direct toxicity for lymphocytes.
Effect of pertussigen on inflammation caused by Freund adjuvant
Authors: Munoz J J , Sewell W A
Published: January 2020

Pertussigen, one of the toxins from Bordetella pertussis, greatly increased the inflammatory response produced by complete Freund adjuvant in the footpads of mice. This effect was not produced by pertussigen when the emulsion was made with saline and incomplete Freund adjuvant, but if an antigen was included in incomplete Freund adjuvant, the strong potentiating effect was again demonstrated. As little as 100 ng of pertussigen given intravenously was effective, but 400 ng proved better, and this latter dose was used routinely. The most striking action occurred when pertussigen was injected on the same day or 3 days after complete Freund adjuvant, but an effect was demonstrated when given from 3 days before to 6 days after complete Freund adjuvant. The action of pertussigen was not apparent until about 6 to 8 days after complete Freund adjuvant. The footpad swelling reached its maximum by day 14 and remained undiminished until day 29. Forty days later, a significant effect was still present. Histologically, the cellular infiltrate in the feet of mice injected with complete Freund adjuvant was more intense in animal treated with pertussigen. Nude BALB/c mice receiving an emulsion of complete Freund adjuvant in the footpads did not respond with an increased inflammation after receiving pertussigen, suggesting the possible involvement of T cells in this phenomenon. The intense and prolonged inflammatory response produced in pertussigen-treated mice by Freund adjuvant containing antigenic substances may serve as a useful model to study chronic inflammation.
Adherence of oral streptococci: evidence for nonspecific adsorption to saliva-coated hydroxylapatite surfaces
Authors: Staat R H , Peyton J C
Published: January 2020

It is proposed that binding of oral streptococci to saliva-coated hydroxylapatite (SHA) surfaces is a multifactorial process involving both specific and nonspecific receptors. In this context, specific binding is described as a high-affinity, saturable interaction between the cell and binding surface. Conversely, nonspecific binding is considered to be a nonsaturable, generalized, low-affinity reaction. Experimental differentiation of specific binding from nonspecific binding was achieved with a competition assay which utilized a large excess of nonradiolabeled bacteria to compete with the 3H-labeled cells for attachment to receptors on 1.5 mg of SHA crystals. Competition assays of Streptococcus sanguis and Streptococcus mitis adhesion clearly demonstrated that the total binding isotherm was composed of a saturable specific binding reaction and a minor nonspecific binding component. This was further substantiated by analysis of nonlinear Scatchard plots of the total binding data. The competition data for Streptococcus mutans binding indicated that ca. 50% of the S. mutans binding appeared to be specific, although saturation of the SHA surfaces with bacterial cells could not be demonstrated. Experiments measuring desorption of radiolabeled cells from SHA crystals into buffer showed that ca. 50% of the bound S. mutans cells were removed after 4 h, whereas less than 5% of the S. sanguis cells were eluted from the SHA surfaces. The kinetics of attachment were studied by using an extract of Persea americana as a noncompetitive inhibitor of adherence. The total cell binding data for these experiments suggested a very rapid binding reaction followed by a slower rate of attachment. It was concluded from these three different experimental approaches that adherence of selected oral streptococci to SHA surfaces involves specific, high-affinity and nonspecific, low-affinity binding reactions. The concept is developed that in vitro streptococcal attachment to SHA can be described as a two-reaction process in which the low-affinity interaction of the cell with the SHA surface precedes the establishment of the stronger, specific bonds needed for the maintenance of streptococci in the oral cavity.
Influence of dose and route of Mycobacterium lepraemurium inoculation on the production of interleukin 1 and interleukin 2 in C57BL/6 mice
Authors: Hoffenbach A , Lagrange P H , Bach M A
Published: January 2020

Groups of C57BL/6 mice were infected either intravenously or subcutaneously with 10(5) or 10(8) Mycobacterium lepraemurium cells, and the ability of their splenic macrophages and T-cells to produce, respectively, interleukin 1 on lipopolysaccharide stimulation and interleukin 2 on concanavalin A stimulation was assessed during the course of infection. In all groups of infected mice, interleukin 1 production remained unaffected during the entire observation period, whereas interleukin 2 activity decreased as the infection progressed. Heavily infected mice (10(8) M. lepraemurium cells) showed an earlier and stronger deficiency interleukin 2 production by concanavalin A-stimulated spleen cells than did mice infected with a lower dose (10(5) bacilli), without detectable influence by the route of inoculation. In mice receiving 10(5) bacilli, minor differences were seen according to the route of infection, with a slight delay in interleukin 2 decrease in mice injected intravenously. In subcutaneously inoculated mice, the failure of spleen cells to produce interleukin 2 after concanavalin A stimulation did not correlate with the number of bacilli developing in the spleen, suggesting the existence of suppressor mechanisms acting at a distance from the site of inoculation.
Inhibition by sugars of Candida albicans adherence to human buccal mucosal cells and corneocytes in vitro
Authors: Collins-Lech C , Kalbfleisch J H , Franson T R , Sohnle P G
Published: January 2020

The adherence and inhibition of adherence of Candida albicans to epithelial cells was studied for human cells obtained from skin (corneocytes) and buccal mucosa. The yeast adhered to both kinds of cells, although in somewhat greater numbers to buccal mucosal cells. Adherence to the cells of different individuals was variable, but the ratios of values for the two kinds of cells from a single subject were quite constant. Inhibition of adherence was produced by several sugars, including the aminosugars mannosamine, glucosamine, and galactosamine. The pattern of inhibition produced by the sugars was similar for the two types of cells. Pretreatment of the yeast with mannosamine, followed by dilution to a subinhibitory concentration, produced some inhibition of yeast-buccal mucosal cell attachment, indicating some direct interaction between the sugar and the fungal cell. These data suggest that the mechanisms whereby C. albicans attaches to corneocytes and to buccal mucosal cells are probably similar.
Effect of splenectomy on production of interferon and colony-stimulating factor in Listeria monocytogenes-infected mice
Authors: Wood P R , Young A M , McKimm-Breschkin J L , Cheers C
Published: January 2020

Splenectomy of mice genetically susceptible to Listeria monocytogenes genetically susceptible mouse strains increased their resistance to L. monocytogenes infection but had no effect on the course of infection in L. monocytogenes-resistant mice. However, splenectomy led to a dramatic decrease in the production of colony-stimulating factor and interferon after L. monocytogenes infection in all mouse strains examined.
Pure paraflagellar rod protein protects mice against Trypanosoma cruzi infection
Authors: Wrightsman R A , Miller M J , Saborio J L , Manning J E
Published: January 2020

The paraflagellar rod proteins (PAR) purified from Trypanosoma cruzi epimastigotes were shown to protect mice against an otherwise lethal challenge inoculum of 10(3) bloodstream-form trypomastigotes. The injection route used for immunization was shown to have a marked impact on the development of protective immunity. Mice receiving subcutaneous (s.c.) injections of PAR proteins had reduced bloodstream parasitemias and showed 100% survival following challenge. In contrast, mice immunized via the intraperitoneal (i.p.) route developed parasitemia levels equivalent to those of unimmunized controls and did not survive infection. Western blotting (immunoblotting) demonstrated that sera from both i.p. and s.c. immunized mice reacted specifically with PAR proteins; however, the antibody titer of the i.p. immunized mice was approximately 64-fold greater than that of the s.c. immunized mice, suggesting that the protective response in the s.c. immunized mice is cell mediated rather than humoral.
Neisserial porins inhibit human neutrophil actin polymerization, degranulation, opsonin receptor expression, and phagocytosis but prime the neutrophils to increase their oxidative burst
Authors: Bjerknes R , Guttormsen H K , Solberg C O , Wetzler L M
Published: January 2020

Porins are trimeric proteins that constitute water-filled pores that allow transmembrane diffusion of small solutes through the outer membrane layer of gram-negative bacteria. The porins are capable of inserting into the membranes of eucaryotic cells, and in the present study we have examined the in vitro effects on neutrophil functions of the following purified porins: meningococcal outer membrane protein classes 1 and 3 and gonococcal outer membrane protein 1B (P1B). The neisserial porins inhibited human neutrophil chemoattractant-induced actin polymerization and degranulation of both primary and secondary granules. The neutrophil expression of immunoglobulin G (IgG) Fc receptors II (Fc gamma RII; CDw32) and III (Fc gamma RIII; CD16), as well as the activation-dependent downregulation of Fc gamma RIII, were reduced by the meningococcal and gonococcal porins. The neisserial porins impaired the upregulation of complement receptors 1 (CD35) and 3 (CD11b) and inhibited the phagocytic capacity of neutrophils, as evaluated by the uptake of meningococci (strain 44/76) in the presence of patient serum containing known amounts of IgG against meningococcal porins. The porins also primed neutrophils to increase their intracellular hydrogen peroxide production in response to FMLP, whereas no such priming was observed if the neutrophil protein kinase C was stimulated directly with phorbol myristate acetate. The neisserial porins influenced neutrophil functions in a time- and concentration-dependent manner. The meningococcal class 1 outer membrane protein and the gonococcal P1B tended to alter neutrophil functions more than the meningococcal class 3 protein. Thus, the neisserial porins inhibited human neutrophil actin polymerization, degranulation, opsonin receptor expression, and phagocytosis but primed the neutrophils to increase their oxidative burst. It remains to be determined whether these in vitro observations reflect mechanisms that may be of importance for the interaction between neutrophils and Neisseria species in vivo.
Effect of enteropathogenic Escherichia coli on adherent properties of Chinese hamster ovary cells
Authors: Vanmaele R P , Finlayson M C , Armstrong G D
Published: January 2020

Enteropathogenic Escherichia coli (EPEC) O111:H2, O119:H6, or O142:H6 caused rapid detachment of Chinese hamster ovary (CHO) cell monolayers within 2 to 4 h of cocultivation. CHO cell detachment was not promoted by nonenteropathogenic E. coli (O125:H4, O126:H27, O157:H7, and O26:H11) and could not be attributed to EPEC production of enterohemolysin or Shiga-like toxins. In contrast, EPEC strains did not promote rapid detachment of Lec1, Lec2, or Lec8 CHO cell monolayers. These CHO cell Lec mutants all express abbreviated glycan sequences on membrane glycoproteins and glycolipids. Although EPEC strains failed to alter the adherent properties of Lec2 cells lacking only terminal sialic acid groups, EPEC adherence to the Lec2 mutant was indistinguishable from that observed with wild-type CHO cells. There was also no significant difference in EPEC-induced actin accumulation or invasion of Lec2 cells. In contrast, EPEC localized adherence to Lec1 and Lec8 mutants, lacking sialyllactosamine (Lec1) or sialic acid and galactose (Lec8) sequences, was reduced by 84 and 93%, respectively. Our results suggest that lactosamine sequences [beta Gal(1-4 or 1-3)beta GlcNAc] not containing sialic acid are sufficient for EPEC adherence, actin accumulation, and invasion of CHO cells. Sialic acid groups, however, may be necessary for EPEC-mediated CHO cell detachment.
Reactivation of persistent Chlamydia trachomatis infection in cell culture
Authors: Beatty W L , Morrison R P , Byrne G I
Published: January 2020

Gamma interferon induces persistent chlamydial infections in cell culture. These infections are characterized by altered morphologic and biochemical features of the pathogen. These persistent forms are abnormally large and noninfectious and undergo unusual structural and functional changes, including production of a paucity of outer envelope constituents and normal levels of the chlamydial hsp60, an immunopathological antigen. The current investigation evaluates the events that occur during reactivation of infectious Chlamydia trachomatis from persistently infected cell cultures. Transfer of persistent chlamydial organisms to gamma interferon-free medium resulted in recovery of infectivity accompanied by an increase in levels of structural membrane proteins and reorganization of aberrant organisms to morphologically typical elementary bodies. In addition, reactivation of infectious organisms from persistent chlamydiae that were maintained in culture for several weeks was demonstrated. These studies show that persistent C. trachomatis maintains viability for extended periods, illustrate the reversibility of immunologically mediated persistent infections, and characterize reactivation at the ultrastructural and biochemical levels.
Expression of the Arp protein, a member of the M protein family, is not sufficient to inhibit phagocytosis of Streptococcus pyogenes
Authors: Husmann L K , Scott J R , Lindahl G , Stenberg L
Published: January 2020

Many Streptococcus pyogenes immunoglobulin-binding proteins have structural similarities to the antiphagocytic M protein, including the well-known C repeats. One of these molecules is the immunoglobulin A-binding protein Arp, which is expressed by a serotype 4 strain for which no antiphagocytic M protein has yet been described. We expressed Arp4 in an S. pyogenes strain from which the structural gene for the M protein has been deleted and found that Arp4 is not sufficient to inhibit phagocytosis.
Pseudomonas aeruginosa infection of the cornea and asialo GM1
Authors: Zhao Z , Panjwani N
Published: January 2020

Extensive immunohistochemical and thin-layer chromatogram-immunostain analyses were carried out to establish whether asialo GM1, a glycolipid which contains binding sites for Pseudomonas aeruginosa, is present in corneal epithelium. The data suggest that rabbit corneal epithelium does not contain detectable levels of asialo GM1 even after corneas are scarified and incubated with trypsin, P. aeruginosa, or P. aeruginosa exoproducts to expose potential cryptic sites. Preliminary immunohistochemical analyses indicated that asialo GM1 is also not found in human corneas.
Rapid disruption of epithelial barrier function by Salmonella typhimurium is associated with structural modification of intercellular junctions
Authors: Jepson M A , Collares-Buzato C B , Clark M A , Hirst B H , Simmons N L
Published: January 2020

Short-term infection of MDCK II monolayers with Salmonella typhimurium SL1344 caused a progressive decrease in transepithelial electrical resistance concomitant with decreased cation permselectivity and increased paracellular inulin flux. Cytochemical staining of F-actin, E-cadherin, and ZO-1 revealed the concentration of each junctional protein in invaded cells as a result of contraction at their apical poles and resultant distortion of adjacent uninvaded cells.
Molecular cloning of the cDNA and gene for an elastinolytic aspartic proteinase from Aspergillus fumigatus and evidence of its secretion by the fungus during invasion of the host lung
Authors: Lee J D , Kolattukudy P E
Published: January 2020

Hydrolysis of structural proteins in the lung by extracellular proteinases secreted by Aspergillus fumigatus is thought to play a significant role in invasive aspergillosis. This fungus was found previously to secrete an elastinolytic serine proteinase and a metalloproteinase. We report that A. fumigatus also secretes an aspartic proteinase (aspergillopepsin F) that can catalyze hydrolysis of the major structural proteins of basement membrane, elastin, collagen, and laminin. The pH optimum for the enzymatic activity was 5.0 with elastin-Congo red as the substrate, and the activity was not significantly inhibited by pepstatin A, diazoacetyl norleucine methylester, and 1,2-epoxy-3-(p-nitrophenoxy) propane. The cDNA and gene encoding this aspartic proteinase were cloned and sequenced. The open reading frame, interrupted by three introns, would encode a protein of 393 amino acids composed of a putative 21-amino-acid signal peptide and a 49-amino-acid propeptide preceding the 323-amino-acid mature protein. The amino acid sequence of A. fumigatus aspartic proteinase has 70, 66, and 67% homology to the sequences of those from Aspergillus oryzae, Aspergillus awamori, and Aspergillus saitoi, respectively. The active-site motif (DTG) and the catalytic aspartic residues characteristic of aspartic proteinases are found in the presently described enzyme, indicating that it belongs to a family of aspartic proteinases. Polyclonal antibodies were produced in rabbits against both the mature and precursor forms of the aspartic proteinase expressed in Escherichia coli. Immunoblotting with both antibodies detected a 39-kDa mature enzyme in the culture supernatant of A. fumigatus. The aspartic proteinase activity was inhibited by the antibodies, suggesting that the aspartic proteinase in the culture supernatant corresponds to the product of the cloned gene. Immunogold electron microscopy showed that the aspartic proteinase was secreted by A. fumigatus invading neutropenic mouse lung and its secretion was directed toward the germ tubes of penetrating hyphae.
Isolation and characterization of fibronectin-binding sites of Borrelia garinii N34
Authors: Kopp P A , Schmitt M , Wellensiek H J , Blobel H
Published: January 2020

Adherence of bacteria to host cell membranes is one of the initial steps of microbial pathogenicity. Numerous studies have suggested that fibronectin promotes this interaction in some bacterial species. In this study, we have examined the ability of Borrelia garinii to bind fibronectin. The binding of fibronectin to the spirochete was specific and saturable. Scatchard plot analysis of the binding data revealed two types of ligands on the spirochetal surface, one with high affinity and one with low affinity for fibronectin. The fibronectin-binding sites were solubilized from the surface of B. garinii N34 by lysozyme treatment. Fast protein liquid chromatography (FPLC) purification of the solubilized binding sites resulted in one band with a high fibronectin-binding activity and a molecular weight of ca. 147,000. FPLC-purified binding sites, fibronectin, and antibodies to fibronectin inhibited the adherence of the spirochete to epithelial cells competitively. These data provide strong support for the hypothesis that fibronectin-binding sites on the surface of B. garinii are involved in the adherence of the spirochete to their respective host cells.
Evidence for apoptosis of murine macrophages by Actinobacillus actinomycetemcomitans infection
Authors: Kato S , Muro M , Akifusa S , Hanada N , Semba I , Fujii T , Kowashi Y , Nishihara T
Published: January 2020

The gram-negative bacterium Actinobacillus actinomycetemcomitans is considered an important etiological agent in periodontal diseases. In this study, we show that A. actinomycetemcomitans strains are cytotoxic for the murine macrophage cell line J774.1. On the other hand, Porphyromonas gingivalis strains, other gram-negative oral species implicated in adult periodontitis, showed weak cytotoxic effects. For this to occur, A. actinomycetemcomitans had to gain entry into the macrophages, since cytotoxicity was prevented by cytochalasin D. We demonstrate that cell death induced by A. actinomycetemcomitans Y4 occurs through apoptosis, as shown by changes in nuclear morphology, an increase in the proportion of fragmented DNA, and the typical ladder pattern of DNA fragmentation indicative of apoptosis. We further sought to determine whether the cytotoxicity induced by A. actinomycetemcomitans Y4 could be modulated by the protein kinase inhibitors H7 and HA1004. Apoptotic cell death induced by A. actinomycetemcomitans Y4 was suppressed by H7 but was relatively unaffected by HA1004. These findings suggest that the signals of protein kinases may regulate apoptosis induced by A. actinomycetemcomitans Y4. The ability of A. actinomycetemcomitans to promote the apoptosis of macrophages may be important for the initiation of infection and the development of periodontal diseases.
A novel costimulatory factor for gamma interferon induction found in the livers of mice causes endotoxic shock
Authors: Okamura H , Nagata K , Komatsu T , Tanimoto T , Nukata Y , Tanabe F , Akita K , Torigoe K , Okura T , Fukuda S
Published: January 2020

Administration of monoclonal anti-CD3 antibody to mice treated with Propionibacterium acnes induced secretion of a high level of gamma interferon (IFN-gamma) into the circulation system, while it induced no significant release in untreated mice. In order to analyze this high-level induction of IFN-gamma in these bacterium-treated mice, we investigated the factors that might be involved. An activity that induces IFN-gamma in T cells was observed in the liver extracts of mice treated with P. acnes and subsequently challenged with lipopolysaccharide. Here, we purified an IFN-gamma-inducing factor from the liver extract to homogeneity and characterized it. Its molecular mass was 18 to 19 kDa, and its pI was 4.9. The amino acid sequence of the NH2-terminal portion was determined and shown to have no similarities to any protein in the EMBL, GenBank, and PIR data bases. The same molecule was also demonstrated in the serum factor that was previously reported to have an IFN-gamma-inducing activity and to have an apparent molecular mass of 75 kDa. Moreover, the activity of this serum factor was recovered in the fraction containing the 18- to 19-kDa protein under reducing conditions and was shown to have the same NH2-terminal amino acid sequence as that of the factor from the liver extract. In addition to the ability to induce IFN-gamma, this protein augmented T-cell proliferation and NK activity in the spleen cells. Thus, several of its biological activities were apparently similar to those of interleukin-12. These results indicated that this novel protein, which exhibited marked costimulatory activity on IFN-gamma production in vitro, was elevated vivo in response to P. acnes treatment. This factor, probably released from the producing cells by lipopolysaccharide stimuli, may be involved in the high-level induction of IFN-gamma in the P. acnes-treated mice.
Proteases of Treponema denticola outer sheath and extracellular vesicles
Authors: Rosen G , Naor R , Rahamim E , Yishai R , Sela M N
Published: January 2020

Electron microscopical observations of the oral periodontopathogen Treponema denticola show the presence of extracellular vesicles bound to the bacterial surface or free in the surrounding medium. Extracellular vesicles from T. denticola ATCC 35404, 50 to 100 nm in diameter, were isolated and further characterized. Protein and proteolytic patterns of the vesicles were found to be very similar to those of isolated T. denticola outer sheaths. They were enriched with the major outer sheath polypeptides (molecular sizes, 113 to 234 kDa) and with outer sheath proteases of 91, 153, 173, and 228 kDa. These findings indicate that treponemal outer sheath vesicles contain the necessary adhesins and proteolytic arsenal for adherence to and damage of eucaryotic cells and mammalian matrix proteins. The major outer sheath- and vesicle-associated protease of T. denticola ATCC 35404 was purified and characterized. The purified enzyme had a molecular size of 91 kDa, and it dissociated into three polypeptides of 72, 38, and 35 kDa upon heating in the presence of sodium dodecyl sulfate with or without a reducing agent. The activity of the enzyme could be inhibited by diisopropylfluorophosphate, phenylmethylsulfonyl fluoride, and phenylboronic acid. The value of the second-order rate constant of the protease inactivation by phenylmethylsulfonyl fluoride was 0.48 x 10(4) M(-1) min-1. Inhibition of the enzyme by phenylboronic acid was rapid (< 1 min) and pH dependent. These data strongly suggest that this major surface proteolytic activity belongs to a family of serine proteases.