bluuebunny/crossref_metadata_2025 huggingface dataset:
Enteropathogencity of
Escherichia coli
I. Evaluation of Mouse Intestinal Loops
Authors: Punyashthiti Kannikar , Finkelstein Richard A.
Published: January 2020

A ligated mouse intestinal loop assay has been introduced to evaluate the enteropathogenicity of strains of
Escherichia coli
. When inoculated as whole cultures, 19 (34%) of 55 strains isolated from diarrheic infants, each of 4 strains associated with diarrhea in human adults, and 5 of 7 strains associated with porcine coli bacillosis were found capable of causing fluid accumulation in this model. In contrast, only 1 of 28 strains isolated from normal individuals was positive in this test. Sterile concentrated filtrates of positive strains cultivated in Brain Heart Infusion broth or syncase broth were also positive, whereas filtrates of negative strains did not cause fluid accumulation. All of the strains associated with diarrhea in human adults and 16 of the 19 positive strains from diarrheic children were also found capable of causing hemagglutination of chicken erythrocytes in a slide test used to evaluate the “adhesiveness” of the bacteria, a property which may enable them to stick in the small bowel and avoid the normally efficient peristaltic clearance mechanism. Few of the nonenteropathogenic strains were positive in this test. The slide hemagglutination test may be a useful, rapid means of screening potentially enteropathic
E. coli
strains and merits further evaluation and study. Although not ideal, the mouse intestinal loop model offers some advantages over the rabbit ileal loop assay.
Interaction of
Actinomyces
Organisms with Cationic Polypeptides I. Histochemical Studies of Infected Human and Animal Tissues
Authors: Crawford James J.
Published: January 2020

Histochemical techniques were used to study the nature of acidophilic hyaline clubs arranged radially at the peripheries of
Actinomyces
colonies in infected lung tissues of two persons. Concentrations of arginine-rich polypeptides were demonstrated in the acidophilic areas and in the cytoplasm of granulocytic leukocytes surrounding the colonies. Exposure of
Actinomyces
organisms to strongly cationic polypeptides (protamine, histone) in vitro killed the organisms and caused them to develop acidophilic staining. Weakly cationic proteins, ribonuclease, and hemoglobin produced no such effects. No acidophilic component could be detected in fresh broth-grown organisms themselves. Viable and nonviable colonies of the test strain lacking hyaline clubs were injected beneath the skin of guinea pigs. Agrinine-rich cationic polypeptides were evident in the cytoplasm of surrounding leukocytes and permeating the microbial colonies. In light of current evidence pertaining to leukocyte lysosomes and capsule production by
Actinomyces
and related organisms, the acidophilic hyaline clubs observed in human tissues appear to be a combination of a capsular component of the actinomycete and a cationic polypeptide component of host leukocytes. Organisms deeper in the human tissue colonies retained their normal basophilic reaction, suggesting a protective role for the peripheral hyaline club matrix. The acidophilic club complexes serve to indicate the reaction of cationic polypeptides in response of the human host to infecting
Actinomyces
organisms. These observations also support a broader concept that antimicrobial polypeptides of leukocyte lysosomes are an important factor in response of both the human and animal host to infecting bacteria.
Characterization of the Group Antigen of
Chlamydia trachomatis
Authors: Dhir Sanittar P. , Kenny George E. , Grayston J. T.
Published: January 2020

A lipid complement-fixing group antigen is shared by all chlamydiae including trachoma organisms. A water-soluble polysaccharide antigen was obtained by alkali saponification from partially purified water-insoluble lipid antigen preparations made from three trachoma strains. The extracted antigen failed to fix complement with antibody but was capable, as a hapten, of inhibiting complement fixation by prior reaction with the antibody. This antigen diffused readily in agarose, producing a reaction of identity with several trachoma strains. The antigen could also be absorbed onto untreated rabbit red blood cells for passive hemagglutination. It is concluded that polysaccharide antigen is responsible for the group reactivity of these organisms and can be obtained from the lipid antigen by alkali saponification.
Characterization of interferons induced by bacteria and interferon-producing leukocytes in human peripheral blood
Authors: Rönnblom L , Forsgren A , Alm G V
Published: January 2020

All of 23 different preparations of formaldehyde-fixed and heat-killed bacteria induced the appearance of high levels of interferon (IFN) in cultures of human peripheral blood mononuclear leukocytes. Some bacteria induced peak IFN titers after 24 h of culture, whereas other bacteria showed maximal titers on culture days 2 to 3. The IFN displayed various properties. One type, which appeared early during the cultures, had characteristics of IFN-alpha, being resistant to pH 2 treatment but neutralized by antibodies to IFN-alpha. A second type, which appeared later, on culture days 2 to 3, resembled IFN-gamma in being sensitive to pH 2 treatment but resistant to anti-IFN-alpha antibodies. A third type, which appeared to be sensitive to both pH 2 and antibody treatment, was interpreted as atypical IFN-alpha. The application of cell fractionation procedures indicated that nonadherent, predominantly Fc receptor-bearing, non-T, non-B cells were producers of IFN-alpha as defined by its antigenic properties. They copurified approximately with cells carrying natural killer activity toward human erythroid leukemia K562 cells. Some bacteria apparently also stimulated T lymphocytes to produce material with properties of IFN-gamma.
Differential toxicity of inhaled gram-negative bacteria
Authors: Baseler M W , Fogelmark B , Burrell R
Published: January 2020

The toxicity by inhalation of various gram-negative bacteria, isolated from settings associated with inhalation disease, was studied by a variety of means. These microorganisms were not equally toxic. Citrobacter freundii aerosol challenges of rabbits provoked significant (up to fivefold) increases in plasma haptoglobin 24 to 48 h after inhalation. Other strains tested failed to provoke such statistically consistent increases. Measurements of C-reactive protein in these same animals did not lead to as reliable results, due to the variability of the responses. Mice responded differently to inhalation in that haptoglobin responses were either unaffected or depressed. When a strain of mice was used that exhibits more severe inflammatory responses to endotoxin (C3H/HeJ), C. freundii and Escherichia coli aerosols provoked significant haptoglobin increases. Free lung cell analyses demonstrated that the macrophage and neutrophil responses differed depending on the strain of bacteria used. Again, C. freundii induced the greatest responses. When murine B lymphocytes were stimulated with lipopolysaccharide preparations from different gram-negative bacteria, distinctly different dose-response curves were obtained. The types of responses obtained indicate that (i) brief inhalation of bacterial aerosols previously thought to be innocuous may lead to pulmonary inflammation, and (ii) that these bacteria differ in their toxicity, with C. freundii being the most toxic organism of the five studied.
Differential activation of resident macrophage subsets with two sources of lymphokine preparations
Authors: Tabor D R , Saluk P H
Published: January 2020

Resident murine macrophages were separated into subsets by Percoll density gradient centrifugation before treatment with lipopolysaccharide-stimulated lymphocytes or different lymphokine preparations. The lymphokines used were culture supernatants from lymphocytes obtained from lipopolysaccharide-injected mice or from purified protein derivative-treated lymphocytes from mice bearing an active BCG infection. The macrophage subsets were activated by the stimulated lymphocytes or lymphokine preparations to express C3b receptor-mediated ingestion or to inhibit the intracellular replication of Toxoplasma gondii or both. The results showed that the macrophage subsets were heterogeneous with respect to ingestion and T. gondii inhibition when activated with lipopolysaccharide-stimulated lymphocytes or lipopolysaccharide-derived lymphokines but were all homogeneous when activated with lymphokines from purified protein derivative-stimulated lymphocytes. When the macrophage subsets were allowed to remain in vitro for various times before lymphokine treatment, the relative pattern of subset activation changed when treated with lipopolysaccharide-derived lymphokines. In contrast, the macrophage subsets remained equally activated throughout the in vitro period when treated with the lymphokines from purified protein derivative-stimulated lymphocytes. The results suggested that functional macrophage heterogeneity depends not only on the nature of the activating signal but also on a state of receptivity of that signal by the macrophage population.
Importance of complement source in bactericidal activity of human antibody and murine monoclonal antibody to meningococcal group B polysaccharide
Authors: Zollinger W D , Mandrell R E
Published: January 2020

The bactericidal activity of human antibody and murine monoclonal antibody to meningococcal group B polysaccharide was investigated as a function of the complement source. The immunoglobulin M murine monoclonal antibody 2-2-B was shown by several different methods to be highly specific for meningococcal group B and Escherichia coli K1 capsular polysaccharides. It had strong bactericidal activity in conjunction with either rabbit or human complement, but gave a higher titer with rabbit complement. A strong prozone was observed in each case. Human postvaccination antibody to meningococcal group B polysaccharide was strongly bactericidal with rabbit complement, but had little or no bactericidal activity in conjunction with human complement. Antibodies in adult normal human sera that were bactericidal with rabbit complement were also found to be predominantly directed against the meningococcal group B capsular polysaccharide. Human antibodies that were bactericidal with human complement appeared to be primarily directed against noncapsular antigens.
Corncob formation between Fusobacterium nucleatum and Streptococcus sanguis
Authors: Lancy P , Dirienzo J M , Appelbaum B , Rosan B , Holt S C
Published: January 2020

Corncob formation in dental plaque was believed to be limited to strains of Bacterionema matruchotii and Streptococcus sanguis. We observed recently that strains of Fusobacterium nucleatum also interacted with S. sanguis to form corncobs. Since the fusobacteria are among the first anaerobic filaments to colonize subgingival plaque, these interactions could serve as a connecting link between the transformation of supra- to subgingival plaque. To further characterize these interactions, quantitative in vitro studies of the kinetics of corncob formation of the fusobacteria were undertaken. These studies indicated that fewer streptococci were needed to saturate F. nucleatum strain 364 compared to strain 10953. Corncob formation with both strains was enhanced with increasing pH up to pH 8, at which point autoaggregation of the streptococci occurred. Variation in ionic strength and divalent cations had little effect on the interaction, and EDTA suppressed aggregate formation only slightly. Detergents at concentrations above 0.05% also inhibited corncob formation. Electron micrographs suggested that attachment of the cocci to the fusiforms was mediated through localized tufts of fimbriae, as they are in the Bacterionema system. However, although both trypsin and heat treatment of the streptococci inhibited corncob formation with fusobacteria, the effects were not as complete as those seen in Bacterionema species. Unlike the Bacterionema model, trypsin and heat treatment of the fusobacteria resulted in inhibition of corncob formation. These results suggest that several different receptors may be involved in corncob formation.
Susceptibility of congenitally athymic (nude) mice to sporotrichosis
Authors: Dickerson C L , Taylor R L , Drutz D J
Published: January 2020

Congenitally athymic (nu/nu) mice were found to be more susceptible to intravenous challenge with Sporothrix schenckii than their phenotypically normal (nu/+) littermates as measured by lethality and the number of viable yeast cells in the liver 7 days postinfection. Thymus reconstitution of nu/nu mice (nu/thy) conferred a significant degree of resistance to sporotrichosis. Immunization greatly enhanced the resistance of nu/thy and nu/+ mice, but unexpectedly increased the susceptibility of nu/nu mice. The susceptibility of nonimmunized nu/nu mice and the finding that thymus transplants augmented resistance to sporotrichosis suggest that T lymphocytes are critical to host defense.
Hydrophobicity and adherence of oral streptococci after repeated subculture in vitro
Authors: Westergren G , Olsson J
Published: January 2020

Fresh isolates of Streptococcus mutans, Streptococcus sanguis, and Streptococcus salivarius from human dental plaque were all highly hydrophobic. After repeated subculture in vitro on blood agar, strains of S. mutans serotype c showed decreased hydrophobicity, whereas serotype d/g strains did not. Parallel to the decreased hydrophobicity in the serotype c strains, an impaired ability to adhere to hydroxyapatite was observed. A similar but less pronounced decrease in hydrophobicity in one S. sanguis strain resulted in a marked decrease in adherence to hydroxyapatite.
Generation of hydrogen peroxide by Candida albicans and influence on murine polymorphonuclear leukocyte activity
Authors: Danley D L , Hilger A E , Winkel C A
Published: January 2020

Iodide fixation by murine polymorphonuclear leukocytes (PMN) incubated with viable Candida albicans blastoconidia increases directly with yeast cell concentration up to about 3 x 10(6) cells per ml, but above this concentration bound activity declines dramatically. To understand the basis for this decline, we examined the oxidative metabolism of fungi and stimulated PMN and found some remarkable similarities between these cell types. Both produced 14CO2 when incubated with [1-14C]glucose, both reduced cytochrome c, and both fixed radiolabeled iodide, although the fungi required exogenous lactoperoxidase. In dose-response experiments, iodination by fungi with lactoperoxidase was identical to that with PMN, i.e., the maximum bound activity occurred in cultures with 10(6) to 3 x 10(6) blastoconidia per ml. Iodination by fungi with lactoperoxidase was reduced when blastoconidia were incubated at 25 degrees C or in the presence of catalase and the metabolic inhibitors rotenone, antimycin A, and 2-deoxyglucose. Results from assays for oxidation of scopoletin and o-dianisidine showed that 10(6) blastoconidia in 1.0 ml of medium released 0.5 to 0.7 nmol of H2O2 after 1 h, but 3 X 10(6) and 10(7) cells released significantly less H2O2. These results suggest that iodide fixation by PMN and low numbers of fungal cells may reflect a cooperative effort, with fungi generating some H2O2 that reacts with the myeloperoxidase released from the PMN. With high concentrations of blastoconidia, H2O2 activity appeared to be specifically inhibited, possibly to protect fungal cells from damage.
Occurrence of plasmid DNA in serologically defined strains of Campylobacter jejuni and Campylobacter coli
Authors: Bradbury W C , Marko M A , Hennessy J N , Penner J L
Published: January 2020

Forty Campylobacter jejuni and 17 Campylobacter coli strains that constitute the set of reference strains for our serotyping scheme were each examined for the presence of plasmid DNA. Agarose gel electrophoresis of alkaline-extracted DNA showed the occurrence of 29 bands in 11 C. jejuni strains and 40 bands in C. coli strains. Plasmids ranged in size from 1.6 to 70 megadaltons. Most strains that carried plasmids had between 2 and 6 of them; however, one strain had 14 plasmids, and two strains contained only 1 plasmid each. Repeated electrophoresis demonstrated that all plasmid profiles were stable. A different plasmid profile was seen for each of the 19 plasmid-carrying strains, but it was clear that plasmids of the same or similar molecular weight could be found in different strains. On the basis of these findings, we are persuaded that plasmid profiles determined by a rapid procedure for DNA extraction will play a significant role in resolving complexities among strains that are difficult to serotype and could be useful in epidemiological studies in which the implicated isolates are plasmid bearers.
Inhibition of Chlamydia psittaci in oxidatively active thioglycolate-elicited macrophages: distinction between lymphokine-mediated oxygen-dependent and oxygen-independent macrophage activation
Authors: Byrne G I , Faubion C L
Published: January 2020

Immune sensitization of spleen cells was required to generate lymphokines (LK) that activated thioglycolate-elicited peritoneal macrophages (thio MACs) to respond via both oxygen-dependent and oxygen-independent systems. LK produced by incubating spleen cells from immunized A/J and LAF mice with concanavalin A stimulated a response by thio MACs to phorbol-12-myristate-13-acetate (PMA)-induced chemiluminescence and activated these cells to inhibit intracellular Chlamydia psittaci replication. Concanavalin A-incubated spleen cell preparations from unimmunized animals stimulated neither PMA-induced chemiluminescence nor antichlamydial activity. Activated thio MACs demonstrated a rapid chemiluminescence response to the intracellular protozoan Toxoplasma gondii, but C. psittaci did not induce chemiluminescence in LK-activated thio MACs, although cells exposed to C. psittaci retained their responsiveness to PMA-induced chemiluminescence. The PMA-induced response was inhibited by the addition of exogenous superoxide dismutase and catalase and was therefore related to the production of superoxide anion (O2 . -) and H2O2 by these cells. LK preparations incubated at 56 degrees C before macrophage treatment retained antichlamydial activity, but heated preparations no longer stimulated thio MACs to respond in the chemiluminescence assay. These data provide evidence that macrophage oxygen-dependent and oxygen-independent systems are simultaneously activated by LK, and these preparations comprise at least two distinct activities. The portion responsible for activating oxygen-dependent systems (PMA-induced chemiluminescence) is heat labile, whereas the portion responsible for activating oxygen-independent systems is heat stable. It is the latter system that results in restriction of chlamydial growth and in vitro parasite persistence.
Relationship of progesterone- and estradiol-binding proteins in Coccidioides immitis to coccidioidal dissemination in pregnancy
Authors: Powell B L , Drutz D J , Huppert M , Sun S H
Published: January 2020

Pregnancy is a major risk factor for coccidioidal dissemination. Because rates of Coccidioides immitis growth and endospore release are stimulated in vitro by levels of unbound progesterone and 17 beta-estradiol that are achievable, in vivo, in the sera of pregnant women (i.e., 10(-9) to 10(-8) M), a specific-hormone-binding system in C. immitis was sought. Fungal cytosols were incubated with tritiated steroids plus or minus radioinert steroids to identify specific binding systems. All five strains of C. immitis tested exhibited specific saturable binding for progestin, estrogen, androgen, and (to a lesser extent) corticosterone and glucocorticoid hormone classes. Only low or inconsistent estrogen or androgen binding was found in Blastomyces dermatitidis and Torulopsis glabrata. Cryptococcus neoformans, Paracoccidioides brasiliensis, and non-albicans Candida species showed no binding. Scatchard analysis of progestin and estrogen binding in C. immitis revealed a high-capacity, low-affinity binding system that was unaffected by RNase and DNase, but 40 to 60% degraded by trypsin or heating. Ammonium sulfate precipitation resolved a high-affinity, low-capacity binding system (Kd = 1.24 X 10(-9) to 3.60 X 10(-8) M; number of binding sites = 0.014 to 0.20 pmol/mg of protein). The Kd of this system is sufficient to compete for unbound hormone in the sera of pregnant women. The high-capacity, low-affinity system may serve as a repository for hormone before its attachment to the specific binder. These studies suggest that the effects of nanomolar concentrations of sex hormones on C. immitis may be mediated by a specific cytosol protein-binding system and that stimulatory events observed in vitro may have relevance for the mechanism of coccidioidal dissemination in pregnancy.
Protease production by Pseudomonas aeruginosa isolates from patients with cystic fibrosis
Authors: Hastie A T , Hingley S T , Kueppers F , Higgins M L , Tannenbaum C S , Weinbaum G
Published: January 2020

The temporal appearance of extracellular proteases produced by Pseudomonas aeruginosa was analyzed by pH 9 and pH 4 polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate-PAGE. Ammonium sulfate precipitates of culture supernatants from various stages of growth revealed a time-dependent increase in number and amount of proteolytically active proteins. One mucoid P. aeruginosa clinical isolate and its derived nonmucoid variant, as well as two other nonmucoid variant P. aeruginosa strains (all from cystic fibrosis patients), showed similar production of five differently migrating proteases (P1 to P5, numbered according to increasing net negative charge) in pH 9 PAGE and one protease in pH 4 PAGE. P2, P3, and P5 increased to maximum concentrations at 24 to 48 h, decreasing thereafter, whereas P4 continued increasing even at 83 h, and P1 fluctuated. P3 was identified as an elastase. P2 was possibly composed of polypeptide chains bridged by disulfide bonds, since without reduction it migrated in sodium dodecyl sulfate-PAGE as a single protein, and with reduction it migrated as three protein bands. Two-dimensional PAGE revealed multiple molecular weight species within protease-positive bands in pH 9 gel strips. Isoelectric focusing gave a pattern of protein separation that correlated with two-dimensional PAGE analysis. Thus, greater heterogeneity of active proteases than previously reported has been demonstrated in all P. aeruginosa clinical isolates studied by sensitive two-dimensional PAGE analysis.
Immunogenicity of a low-virulence vaccinal strain against Salmonella abortus-ovis infection in mice
Authors: Lantier F , Pardon P , Marly J
Published: January 2020

Strain Rv6, a streptomycin-independent reverse mutant of Salmonella abortus-ovis, multiplied and disseminated in vivo to a lesser extent than the virulent parental strain 15/5, as determined by subcutaneous inoculation of two outbred strains of mice with different susceptibilities to salmonella infection. This lower virulence of strain Rv6 was not modified by 10 passages in susceptible mice. Vaccination with strain Rv6 induced a dose-dependent capacity of mice to control subsequent infection with the virulent parental strain. Induced resistance began as early as 3 days after vaccination and lasted at least 11 months. Its efficiency was controlled by different routes and doses of challenge. Strain Rv6 induced a greater protection than that conferred by heat-killed bacteria or by tested commercial killed vaccines.
Rapid induction of hypothyroidism by an avian leukosis virus
Authors: Carter J K , Smith R E
Published: January 2020

Infection of 10-day chicken embryos with an avian leukosis virus, RAV-7, resulted in hypothyroidism within 3 weeks posthatching. Histological examination of the thyroids from infected chickens showed an extensive infiltration of lymphoblastoid cells by 7 days posthatching. Areas resembling germinal centers were present in the thyroids of infected chickens by 3 weeks posthatching. Examination of circulating thyroid and pancreas hormones showed a significant reduction in T3 and T4 levels and a trend toward higher insulin levels after 16 days posthatching. T4 supplementation of RAV-7-infected chickens alleviated some aspects of the disease syndrome but did not abrogate all symptoms. Marked involution of both bursa and thymus glands was noted. RAV-7 had an RNA genome of 8.2 kilobases and a polypeptide composition characteristic of an avian leukosis virus. The hypothyroidism followed a dose response to RAV-7 infection.
Activation of macrophages for microbicidal and tumoricidal effector functions by soluble factors from EL-4, a continuous T cell line
Authors: Nacy C A , James S L , Benjamin W R , Farrar J J , Hockmeyer W T , Meltzer M S
Published: January 2020

Macrophages treated with culture fluids from EL-4 cells, a continuous T cell line, were activated to kill mKSA-TU-5 fibrosarcoma cells, amastigotes of Leishmania tropica, and schistosomula of Schistosoma mansoni. Active EL-4 factors eluted from Sephadex G-100 in two distinct regions: molecular weight 45,000 (activities induced killing of unrelated intracellular and extracellular targets) and molecular weight 23,000 (activities induced killing of extracellular targets only). These results confirm heterogeneity among activation signals for the induction of macrophage microbicidal and tumoricidal activities. Factors that induced cytocidal activity against extracellular tumor cells and schistosomula were distinct from those that induced destruction of intracellular amastigotes.
Surface antigens of metacyclic trypomastigotes of Trypanosoma cruzi
Authors: Yoshida N
Published: January 2020

The surface antigen makeup of metacyclic trypomastigote forms of strain G of Trypanosoma cruzi, which produce a subpatent infection in mice, differed from those of the virulent strains Y and CL. A 100,000-molecular-weight protein, barely detectable on the Y or CL cell surface, appeared as the main surface antigen of the G metacyclic trypomastigotes. In addition, the G metacyclic forms differed from those of the virulent strains in their susceptibility to complement-mediated immunolysis.
Lysosomal response of a murine macrophage-like cell line persistently infected with Coxiella burnetii
Authors: Akporiaye E T , Rowatt J D , Aragon A A , Baca O G
Published: January 2020

The lysosomal response of a murine macrophage-like tumor cell line (J774) during persistent infection with Coxiella burnettii was examined. By using acid phosphatase as a lysosomal marker, it was shown that phagosome-lysosome fusion occurred in J774 cells persistently infected with C. burnetii. This observation was verified using thorium dioxide, an electron-dense compound that is sequestered in secondary lysosomes. The phagolysosomes contained viable replicating rickettsiae. Spectrofluorometric analysis indicated that the phagolysosomal pH of persistently infected cells was acidic. In attempts to correlate rickettsial survival with lysosome function, the activities of several lysosomal enzymes were assayed in both infected and uninfected cells. Activities of acid phosphatase and beta-acetylglucosaminidase were not significantly altered during infection. However, infected cells appeared to display slightly higher intracellular lysozyme, beta-glucuronidase, and beta-galactosidase activities.
Induction of a novel morphological response in Chinese hamster ovary cells by pertussis toxin
Authors: Hewlett E L , Sauer K T , Myers G A , Cowell J L , Guerrant R L
Published: January 2020

Exposing Chinese hamster ovary cells in culture to pertussis toxin resulted in a novel clustered growth pattern. The specificity of the response for pertussis toxin was shown by neutralization of the activity with specific anti-toxin antibody, heat lability (80 degrees C for 15 min), and absence of such activity by culture media from nontoxigenic Bordetella species. Although a lag of at least 16 h was required before clustered growth was seen, exposure to the toxin for as little as 10 min resulted in a full response 24 h later. The morphological effect appeared to be independent of the cyclic AMP-mediated cell elongation elicited by the heat-labile enterotoxin from Vibrio cholerae in that the pertussis toxin effect was seen in both the presence and absence of elongation. Although the mechanism by which this effect is mediated remains to be determined, it is already providing a useful in vitro assay for pertussis toxin.
A factor from Actinomyces viscosus T14V that specifically aggregates Streptococcus sanguis H1
Authors: Mizuno J , Cisar J O , Vatter A E , Fennessey P V , McIntire F C
Published: January 2020

A highly specific aggregation factor for Streptococcus sanguis H1 (AFH1) was obtained by lysozyme treatment of Actinomyces viscosus T14V. At 1 micrograms/ml, AFH1 aggregated a suspension of S. sanguis H1, with which A. viscosus T14V coaggregates by a mechanism not inhibited by lactose: even at much higher levels AFH1 caused little or no aggregation of streptococci from other coaggregation groups (J. O. Cisar et al., Infect. Immun. 24:742-752, 1979). The most active fraction of AFH1 obtained by gel chromatography (near the void volume of Bio-Gel A1.5 m) reacted as a single antigen with anti-A. viscosus T14V serum and was unrelated to the fimbrial antigens of A. viscosus T14V. Smaller molecular fractions, at high levels, inhibited aggregation of S. sanguis H1 by high-molecular-weight AFH1 as well as coaggregation of S. sanguis H1 with A. viscosus T14V. The AFH1 fraction with high aggregating activity was composed of approximately 53% cell wall components (alanine, glutamine, lysine, N-acetylglucosamine, and N-acetylmuramic acid). 40% polysaccharide (N-acetylgalactosamine, rhamnose, and 6-deoxytalose), and 7% protein; teichoic acid was not detected. The fraction which inhibited aggregation and coaggregation contained much less of the cell wall constituents and more of the polysaccharide than the fraction with potent aggregating activity. Aggregation was completely prevented either by treating AFH1 with 0.01 M periodate at 25 degrees C for 4 h or by treating S. sanguis H1 with heat or pronase. A role for electrostatic forces in the aggregation was indicated by: (i) NaCl inhibition of aggregation, and (ii) a great decrease in aggregation potency as a result of chemical modification of either cationic or anionic groups of AFH1. On the other hand, NaCl reversed the aggregation only very weakly. The overall data suggest that a carbohydrate-protein interaction may be dominant in the aggregation of S. sanguis H1 by AFH1 and in the coaggregation of S. sanguis H1 with A. viscosus T14V.
Protection in rabbits immunized with a vaccine of Escherichia coli heat-stable toxin cross-linked to the heat-labile toxin B subunit
Authors: Klipstein F A , Engert R F , Houghten R A
Published: January 2020

Rabbits and rats were immunized with a vaccine consisting of synthetically produced Escherichia coli heat-stable toxin cross-linked by the carbodiimide reaction to the B subunit of biologically produced porcine heat-labile toxin. The vaccine contained 50% of each toxin component by weight and antigenicity; the toxicity of the heat-stable enterotoxin component was reduced by greater than 600-fold. Two or three peroral immunizations with vaccine containing 1,000 antigen units of each component raised greater-than-threefold increases in specific mucosal immunoglobulin A antitoxin titers to each component in all animal groups. Protection index values for challenge with either heat-labile or heat-stable toxins in ligated ileal loops were 3.4 to 4.0 in rats immunized by a parenteral primary immunization followed by two peroral booster immunizations, greater than 9 in rabbits immunized by these routes, and greater than 8 in rabbits given just three peroral immunizations. The antigenicity of the B-subunit component of the peroral vaccine was protected equally well against gastric acidity either by pretreatment with cimetidine or by delivery of the vaccine encapsulated in pH-dependent microspheres. The vaccine did not cause diarrhea when given perorally to any of the experimental animals or evoke fluid secretion when instilled into rabbit ligated ileal loops. These observations (i) confirm the effectiveness of this vaccine as an immunogen in a second animal model, (ii) establish that it is effective when given exclusively by the peroral route, and (iii) provide further evidence regarding its lack of toxicity.
T and B lymphocyte susceptibility to murine leukemia virus moloney
Authors: Isaak D D , Cerny J
Published: January 2020

The susceptibility of T and B lymphocytes to productive infection and transformation by murine leukemia virus Moloney was determined by enumeration of cells producing infectious virus after in vitro infection of mitogen-stimulated, isolated cell populations and by in vivo infection of euthymic BALB/c and thymus-deficient (nude) mice. Our in vitro results demonstrated that the majority of splenic T cells and thymocytes are resistant to productive infection in vitro; a specific subpopulation of susceptible nylon-adherent splenic T cells was identified, however. Similarly, surface immunoglobulin-positive B cells also represent susceptible targets in vitro; mature B cells, however, did not represent the principal target for transformation in the in vivo experiments. Infected euthymic mice expressed increasing titers of murine leukemia virus and uniformly developed fatal T-cell lymphomas at 10 to 12 weeks postinfection; nude mice, in contrast, maintained high, stable levels of viremia throughout the 28 weeks of observation. Infected nude mice remained free of malignancy or developed either granulocytic leukemias or, in one case, reticulum cell sarcoma. Collectively, the results indicate that while the majority of T cells are resistant to productive infection, they represent the principle targets for transformation; B cells, however, represent permissive targets for virus replication, but are resistant to transformation.
Pathogenesis and immune response of vaccinated and unvaccinated rhesus monkeys to tick-borne encephalitis virus
Authors: Hambleton P , Stephenson J R , Baskerville A , Wiblin C N
Published: January 2020

The rhesus monkey was used as a model for diseases caused by viruses of the tick-borne encephalitis virus complex to study the efficacy and safety of a commercial killed vaccine. Animals infected intravenously developed a subclinical infection with no histopathological lesions but with transient clinical chemical changes that included elevated transaminase, dehydrogenase, and creatine kinase activities and that declined as an immune response developed. The immune response was detected as neutralizing antibody in serum and serum antibody to several viral proteins. Antibodies to viral envelope protein and two other infected cell-specific polypeptides were also detected. Intranasal infection resulted in a disease resembling that in humans, except that no pyrexia was observed. Clinical chemical changes similar to those in intravenously infected monkeys developed, but most animals died before an immune response was mounted. Using this model, we have demonstrated that a commercial vaccine protects animals against a wild-type virus isolate and that it elicits an effective immune reaction without any evidence of an immune enhancement phenomenon or adverse side effects as judged by clinical observation, clinical chemistry, and histopathology.
Biphasic effect of pertussis vaccine on serum insulin in mice
Authors: Hewlett E L , Roberts C O , Wolff J , Manclark C R
Published: January 2020

Administration of pertussis vaccine, consisting of whole, killed Bordetella pertussis organisms, causes hyperinsulinemia and enhanced secretion of insulin in response to a variety of secretagogues in rats and mice. In examining the time course and properties of this phenomenon, we discovered two distinct and separate effects of the bacteria on glucose and insulin levels in mice. First, a heat-stable (80 degrees C for 30 min) component causes a brief hyperinsulinemia which is +measureable by 1 h, maximal at 8 h, and ends in less than 48 h. This effect appears to be due to B. pertussis endotoxin, is mimicked by Escherichia coli endotoxin, and is associated with a transient, mild hypoglycemia. Second, there is a heat-labile component of the B. pertussis organism which induces a sustained (greater than 14 days), dose-dependent hyperinsulinemia which reaches a maximum at 5 to 7 days and has no associated hypoglycemia. The two effects are further distinguishable in that the early, endotoxin-induced hyperinsulinemia exhibits the normal suppressibility by exogenous epinephrine, whereas epinephrine markedly enhances the hyperinsulinemia occurring at 7 days. These two effects of B. pertussis appear to be mediated by different mechanisms and may be important in the well-recognized reactogenicity of pertussis vaccine in humans.
Association of bacterial carbohydrate-specific cold agglutinin antibody production with immunization by group C, group B type III, and Streptococcus pneumoniae type XIV streptococcal vaccines
Authors: Colling R G , Pearson T C , Brown J C
Published: January 2020

Rabbits immunized with group B type III, group C, and Streptococcus pneumoniae type XIV streptococcal vaccines developed autoantibodies reactive with autologous and isologous erythrocytes and human O-positive erythrocytes at reduced temperatures. The cold agglutinin antibodies were present in both the immunoglobulin M (IgM) and IgG fractions of group C streptococcal antiserum and in the IgM fraction of group B type III and S. pneumoniae type XIV antisera. BALB/c, CF1, and local strains of mice immunized with group B type III and S. pneumoniae type XIV streptococcal vaccines also produced a cold agglutinin antibody reactive with rabbit and human erythrocytes. The cold agglutinin antibodies were reactive with saccharide compounds representative of the determinants present on the individual bacterial carbohydrate structures, individual vaccine preparations, and isolated polysaccharides. The group C antibodies in rabbits were reactive with sugar ligands in the following order: N-acetylgalactosamine greater than melibiose greater than lactose greater than galactose greater than glucose. Group B type III and S. pneumoniae type XIV cold agglutinin antibodies in rabbit antisera, however, displayed reactivities different from group C antibodies and from each other. Group B type III antibodies reacted with galactose greater than lactose greater than N-acetylgalactosamine greater than glucose greater than rhamnose; S. pneumoniae type XIV antibodies reacted with lactose greater than melibiose greater than galactose greater than glucose greater than N-acetylgalactosamine. The same relative ligand specificity was observed for the cold agglutinin antibodies in S. pneumoniae type XIV mouse antisera. The cold agglutinin antibodies in group B type III and S. pneumoniae type XIV antiserum reacted with erythrocytes at higher temperatures (up to 31 degrees C) than did group C antibodies (up to 14 degrees C). In addition, S. pneumoniae type XIV antibodies did not discriminate between I- or i-bearing human erythrocytes to a significant extent. The results obtained provide substantial evidence that autoreactive cold agglutinin antibodies produced by immunization with these vaccines represent subpopulations of bacterial carbohydrate-specific antibodies that cross-react with mammalian carbohydrate structures.
Experimental pneumococcal meningitis: role of leukocytes in pathogenesis
Authors: Ernst J D , Decazes J M , Sande M A
Published: January 2020

Two groups of rabbits with experimental meningitis induced by direct intracisternal inoculation of Streptococcus pneumoniae cells were studied. One group was rendered profoundly leukopenic by nitrogen mustard, and the other had normal leukocyte counts. The two groups had comparable bacterial growth rates (mean generation time, 60 versus 67 min) and ultimate bacterial populations in the cerebrospinal fluid (CSF) (mean log10 CFU, 9.1 versus 8.7); therefore leukocytes did not effectively slow or limit the growth of pneumococci in the CSF in vivo. Increased CSF protein, decreased CSF glucose, and increased CSF lactate levels were similar in both groups, suggesting that leukocytes are not essential for these changes to occur. Quantitative blood cultures revealed identical levels of pneumococcal bacteremia until 13 to 14 h after the initiation of infection, when the leukopenic rabbits showed a larger number of pneumococci in the blood, ultimately exceeding the number reached in nonleukopenic rabbits by 100-fold. Leukocytes therefore limit the extent of pneumococcal bacteremia after infection of the CSF despite their lack of effect on the course or the CSF manifestations of experimental meningitis.
Enhanced natural killer cell activity in experimental murine encephalitozoonosis
Authors: Niederkorn J Y , Brieland J K , Mayhew E
Published: January 2020

Spleen cells from mice infected with the protozoan parasite Encephalitozoon cuniculi demonstrated enhanced in vitro cytolysis of YAC-1 lymphoma cells. Selective cell depletion experiments showed that the dominant cell population mediating cytolysis of YAC-1 tumor cells expressed the characteristic phenotype of murine natural killer (NK) cells because (i) pretreatment of spleen cells with anti-asialo GM 1 antiserum plus complement abolished the cytotoxic activity; (ii) augmented cytolysis was found in athymic nude mice; (iii) pretreatment of spleen cells with anti-Thy 1.2 plus complement did not affect the level of cytolysis; and (iv) nylon wool removal of adherent cells did not reduce the augmented cytolysis. The augmented cytolysis peaked 7 days after infection, gradually diminished, and finally returned to control levels by 21 days postinfection. The parasite-induced augmentation of NK cell activity was dose-dependent: inoculation of 10(7) parasites gave maximum enhancement, whereas 10(5) or 10(4) parasites had an insignificant effect on spontaneous NK cell cytolysis. The augmented NK cell cytotoxicity was dependent upon viable parasites; inoculation of killed parasites failed to stimulate a significant increase in spontaneous cytolysis. An active infectious process was an important component of this process. The peak of NK activity in euthymic mice was closely correlated with the active stage of infection, and reduction of NK cell activity coincided with recovery from infection. By contrast, athymic nude mice were unable to control E. cuniculi infections yet maintained persistently elevated NK responses. The present data, along with previous reports, indicate that infection with E. cuniculi evokes transient modulation of host immune functions.
Characterization of Toxoplasma gondii antigens that react with human immunoglobulin M and immunoglobulin G antibodies
Authors: Naot Y , Guptill D R , Mullenax J , Remington J S
Published: January 2020

Studies were performed to define the nature of the Toxoplasma gondii antigens that are recognized by human immunoglobulin M (IgM) and IgG antibodies. Both IgM and IgG antibodies were found to be directed mainly against T. gondii membrane antigens in sera obtained from patients with acute toxoplasmosis. Treatment of the membrane preparation with DNase, RNase, or lipase had no apparent effect on the reactivity of the membrane antigens with IgM and IgG antibodies. Lipids isolated from tachyzoites were not recognized by either IgM or IgG antibodies. Exposure of T. gondii membranes to heat, proteolysis, or oxidation with sodium periodate decreased the reactivity of the membrane preparations with both IgM and IgG antibodies. A preparation of T. gondii proteins and polysaccharides were recognized by both immunoglobulin classes. T. gondii polysaccharides reacted with human IgG antibodies produced during both the acute and chronic phases of the infection. We concluded that, after infection with T. gondii, IgM and IgG antibodies are elicited in response to both protein and carbohydrate constituents of the invading parasite.
Evidence for mucins and sialic acid as receptors for Pseudomonas aeruginosa in the lower respiratory tract
Authors: Ramphal R , Pyle M
Published: January 2020

The nature of the receptors for mucoid and nonmucoid Pseudomonas aeruginosa was investigated by using adherence to injured tracheal epithelium as a model. Bovine submaxillary mucin and crude rat tracheal mucin inhibited the adherence of both types of P. aeruginosa. Among the sugars present in these mucins only N-aceylneuraminic acid inhibited adherence. Inhibition of adherence probably involved the binding of N-acetylneuraminic acid to the bacterial cells and not to the tracheal cells. The mucoid strain appeared to be much more sensitive to inhibition by N-acetylneuraminic acid. Periodate oxidation and cholera filtrate also reduced the adherence of both strains, but Clostridium perfringens neuraminidase treatment did not alter adherence. A nonmucoid isogenic mutant of an unstable mucoid strain was also inhibited by N-acetylneuraminic acid. These data suggest that the receptor for P. aeruginosa is a sialic acid moiety on cell surfaces or in mucins.
Cytotoxic effect of an alpha-hemolytic Escherichia coli strain on human blood monocytes and granulocytes in vitro
Authors: Gadeberg O V , Orskov I , Rhodes J M
Published: January 2020

The alpha-hemolytic Escherichia coli strain C134-73 Hly+ was primarily cytocidal to human blood monocytes and granulocytes in vitro in the presence of fresh autologous plasma. Monocytes and granulocytes underwent marked morphological changes during incubation with the bacteria, and the percentages of intact phagocytes decreased progressively with the time of incubation. The cytotoxic effect increased with the number of bacteria per phagocyte and was produced by log-phase microorganisms only. Neither free hemolysin nor free cytotoxic activity to leukocytes could be detected in the incubation medium if the bacteria were removed from the test system. Bacteria-free culture supernatants containing alpha-hemolysin were cytotoxic to monocytes, granulocytes, and lymphocytes, monocytes and granulocytes being the most sensitive. However, this effect was abolished by fresh autologous plasma. Two nonhemolytic strains, a mutant derivative of C134-73 Hly+ and strain X43, were not cytotoxic. These observations suggest that alpha-hemolysin associated with the bacterial cells may enhance the virulence of E. coli by injuring phagocytes, whereas free alpha-hemolysin may be of minor importance because its cytotoxic effect is neutralized by host plasma.
Activation of caprine arthritis-encephalitis virus expression during maturation of monocytes to macrophages
Authors: Narayan O , Kennedy-Stoskopf S , Sheffer D , Griffin D E , Clements J E
Published: January 2020

Lentiviruses, which cause arthritis-encephalitis and maedi-visna in goats and sheep, respectively, cause persistent infections in these animals. The viruses replicate productively at low levels in macrophages in diseased organs such as the "maedi lung" and nonproductively in other cell types such as leukocytes in peripheral blood. Nonproductive infections become productive during in vitro cultivation of the cells. This study showed that monocytes were the only cells in the peripheral blood leukocytes of an infected animal in which virus was detected and that virus activation occurred only when these cells matured into macrophages. Only a minute fraction of cultured monocytes matured into macrophages, and viral infectivity was associated exclusively with this fraction. Antiglobulin-coated glass wool fragments were lethal for monocyte macrophages because of toxic phagocytosis, but had no effect on B or T lymphocytes. The simultaneous addition of the glass fragments and leukocytes to culture dishes resulted in no macrophage maturation and no virus production. The addition of the fragments to virus-producing macrophages caused the death of the cells and a decline in virus production. Virus production in less avidly phagocytic cells was unaffected by the glass. Thus, although macrophages may be permissive for virus replication, one mechanism for restricted virus expression in vivo may be physiological factors controlling the maturation of these cells.