bluuebunny/crossref_metadata_2025 huggingface dataset:
Mannose-sensitive stimulation of human leukocyte chemiluminescence by Escherichia coli
Authors: Mangan D F , Snyder I S
Published: January 2020

Escherichia coli organisms with mannose-sensitive adherence factors (adhesins) are known to associate with human peripheral leukocytes (WBCs) in vitro in the absence of serum. To determine whether the WBC respiratory burst is activated during the interaction with E. coli, WBC chemiluminescence was measured. E. coli with mannose-sensitive adhesins stimulated a sharp burst of chemiluminescence which peaked 15 to 30 min after the bacteria and WBCs were mixed. Stimulation of chemiluminescence could be abrogated by including 10 mM alpha-methyl-D-mannoside in the test suspension. The addition of alpha-methyl-D-mannoside up to 20 min after the E. coli and WBCs were combined caused a rapid decrease in chemiluminescence. E. coli stimulation of chemiluminescence could not be inhibited by pretreating the WBCs with purified type 1 pili (fimbriae). E. coli lacking mannose-sensitive adhesins failed to stimulate chemiluminescence. The results emphasize the importance of mannose-sensitive adhesins in the association of E. coli with WBCs and suggest that the E. coli-WBC interaction system may be a useful tool for studying the mechanisms involved in the activation of the respiratory burst during phagocytosis.
Bacterial aggregating activity in human saliva: simultaneous determination of free and bound cells
Authors: Golub E E , Thaler M , Davis C , Malamud D
Published: January 2020

Two new assays for saliva-mediated aggregation of oral bacteria have been developed, based on the use of [3H]thymidine-labeled cells. One assay separates free cells from aggregated cells by centrifugation through sucrose, whereas the other utilizes membrane filters (8 micrometers, Nuclepore) to effect the separation. Comparison of these assays with the turbidity method reveals that they are faster (X20 to 40) and require 10 times less saliva and bacteria. The aggregation of Streptococcus sanguis M5, as determined with these assays, is complete in 5 min and is dose dependent on added cells and saliva. The reaction exhibits a temperature optimum of 42 degrees C with no reaction at 0 degrees C. If the pH is reduced to below 5, saliva-dependent aggregation is inhibited. The salivary factor(s) are heat labile, losing 100% of their activity after 100 degrees C, 10 min or 70 degrees C, 30 min.
Effect of metal ions on diphtheria toxin production
Authors: Groman N , Judge K
Published: January 2020

The effect of several metal ions on the production of diphtheria toxin was tested. By using the gel immunodiffusion system for detecting toxin, a wide range of metal ion concentrations was conveniently surveyed. Five divalent cations, Fe2+, Cu2+, Co2+, Ni2+, and Mn2+ inhibited toxin production within a range of concentrations that did not inhibit growth of the producing strain. Growth and toxin production were inhibited at identical concentrations by both Cd2+ and Zn2+, whereas Al3+ and Sr2+ affected neither growth nor toxin production over the range of concentrations tested. The data showed that Fe2+ was the most effective inhibitor on an equivalence basis, followed by Cu2+, Co2+, and Ni2+ in descending order. All eight strains of Corynebacterium diphtheriae chosen from diverse ecological origins responded similarly to all metals at similar concentrations. A mutant strain which produces toxin at Fe2+ concentrations 500 times greater than are inhibitory for the parent strain had simultaneously acquired resistance to inhibitory concentrations of Cu2+, Co2+, Ni2+, and Mn2+. This suggests that there is at least one common point in the activity of all these metal ions, and that toxin may respond broadly to changes in metal ion concentrations in the environment.
Aggregation and Adherence of
Streptococcus sanguis
: Role of Human Salivary Immunoglobulin A
Authors: Liljemark William F. , Bloomquist Cynthia G. , Ofstehage John C.
Published: January 2020

Fourteen freshly isolated strains of
Streptococcus sanguis
were obtained from the dental plaque of five healthy adults. Whole saliva was collected concomitant with the plaque isolates from the five subjects, and a second whole saliva sample was collected 10 weeks later. All possible combinations of the first five saliva samples, the second five saliva samples, and 14 strains of bacteria were tested for aggregation. Of the 140 combinations examined, 108 of 140 (77%) of the strains aggregated with the first saliva samples and 95 of 140 (68%) aggregated with the second saliva samples. Overall, 72% of the strains aggregated with both the first and second saliva samples. Removal of immunoglobulin A (IgA) from these same salivas resulted in 38 of 108 (35%) of the aggregates decreasing in intensity with the first saliva samples and 27 of 95 (29%) of the aggregates decreasing in intensity with the second saliva samples. No aggregates increased in intensity with saliva samples when IgA had been removed. Removal of IgA from saliva also resulted in a mean decrease of 46% in adherence of
S. sanguis
to hydroxyapatite coated with the IgA-deficient saliva. Several strains of
S. sanguis
were shown to aggregate strongly with human salivary and colostral IgA. In addition,
S. sanguis
strain S7 showed a 31% stimulation of adherence to hydroxyapatite precoated with human salivary IgA over the uncoated controls. Stepwise removal of IgA from saliva resulted in a decrease in aggregation intensity from strong (4+) to weak (1+ to 2+). Similarly, the adherence of
S. sanguis
to hydroxyapatite coated with these saliva samples decreased linearly as the salivary IgA was depleted. Alternatively, the addition of a small quantity of salivary IgA (20 μg/ml) to progressively diluted saliva maintained a high level of adherence and strong aggregation until the saliva dilutions reached between 1:8 in the adherence experiments and 1:32 for the aggregations. These data indicate that salivary IgA may play an important role in the microbial ecology of human dental plaque formation.
Experimental infection of rabbit ligated ileal loops with Treponema hyodysenteriae
Authors: Knoop F C
Published: January 2020

An in vivo animal model was used to assess the enteropathogenicity of the etiological agent (Treponema hyodysenteriae) of swine dysentery. Multiple ligated ileal loops, prepared in New Zealand white rabbits, were challenged with either pathogenic (B78 and B204) or nonpathogenic (Pu) isolates of the organism. The pathogenic isolates induced the onset of intestinal fluid accumulation as early as 4 h, with maximal fluid induction at 18 h postchallenge. Gross lesions of the intestinal mucosa, observed in ileal loops of rabbits sacrificed 24 h postchallenge, were characteristic of swine dysentery. Both pathogenic isolates colonized the epithelial surface and eroded the mucosal barrier, as determined by histological and scanning electron microscopic observations. Intestinal fluid accumulation and erosion of the mucosal barrier were not observed in ileal loops exposed to the nonpathogenic isolate (Pu) or to either of the nonviable pathogenic (B78 and B204) isolates. The ability of pathogenic isolates to initiate and produce infection in rabbit ligated ileal loops, which closely resembles the disease in swine, provides a system with which to study experimental swine dysentery.
Partial purification of a specific antigen of Treponema hyodysenteriae
Authors: Baum D H , Joens L A
Published: January 2020

A specific antigen of pathogenic Treponema hyodysenteriae was extracted from lyophilized cells of isolant B169 with hot phenol-water, isolated, and semipurified by starch block electrophoresis.
Susceptibility to adjuvant-induced arthritis among germfree, specific-pathogen-free, and conventional rats
Authors: Kohashi O , Kuwata J , Umehara K , Uemura F , Takahashi T , Ozawa A
Published: January 2020

Germfree F344 rats developed severe arthritis with 100% incidence after a single intradermal inection of either squalane containing 0.5 mg of heat-killed Mycobacterium bovis BCG or a water-in-oil emulsion containing 0.2 mg of peptidoglycan derived from Staphylococcus epidermidis. Conventional F344 rats developed less-severe arthritis with 20% incidence for heat-killed BCG and 0% incidence for peptidoglycan. Specific-pathogen-free rats showed an intermediate susceptibility between germfree and conventional rats. Interestingly, both unimmunized specific-pathogen-free and conventional rats. but not unimmunized germfree rats, showed weak delayed-type hypersensitivity reactions to peptidoglycans derived from either S. epidermidis or Lactobacillus plantarum, suggesting that a bacterial flora may furnish a stimulus for induction of cell-mediated immunity to ubiquitous bacterial peptidoglycans. It is thus possible that although a bacterial flora is not necessary for development of adjuvant arthritis, it may have some suppressive effect on the development of the disease in specific-pathogen-free and conventional F344 rats, possibly through modulation of the immune response.
Comparison of natural killer cells induced by Kunjin virus and Corynebacterium parvum with those occurring naturally in nude mice
Authors: Macfarlan R I , Ceredig R , White D O
Published: January 2020

Natural killer (NK) cells are rapidly elicited in the spleen and peritoneal cavity of mice inoculated intravenously or intraperitoneally with live Kunjin virus, and more slowly in the peritoneal cavity of mice inoculated intraperitoneally with Formalin-inactivated Corynebacterium parvum. NK cells induced by either agent display cytotoxicity for a similar spectrum of syngeneic, allogeneic, and xenogeneic cultured cell lines. By contrast, the cells occurring naturally in the spleen of congenitally athymic (nude) mice show substantially lower NK activity and are cytotoxic for a more restricted range of target cell lines. The distinction suggests that there may be more than one type of NK cell or that activation enhances the cytotoxicity and perhaps broadens the range of target specificity of endogenous NK cells.
Attachment of Escherichia coli to urinary sediment epithelial cells from urinary tract infection-prone and healthy children
Authors: Svanborg-Edén C , Jodal U
Published: January 2020

Escherichia coli isolated from patients with recurrent urinary tract infections were tested for ability to attach to human urinary sediment epithelial cells in vitro. Higher mean capacity to bind bacteria was found for epithelial cells of the patient from whom the E. coli strain had been isolated than for epithelial cells from subjects without a history of urinary tract infection. The two populations were age matched. No relation was found between the age of the cell donor (0 to 15 years) and the capacity for E. coli attachement.
Virulence of temperature-sensitive mutants of Sindbis virus in neonatal mice
Authors: Barrett P N , Atkins G J
Published: January 2020

The virulence in neonatal mice of temperature-sensitive (ts) mutants of Sindbis virus was determined by measurements of mean survival time and 50% lethal dose after intracerebral injection. For 11 ts mutants, mean survival time was determined by the ribonucleic acid (RNA) phenotype, RNA+ mutants killing the mice sooner than RNA- mutants for the same titer of virus injected. Mortality caused by seven ts mutants was, with one exception, correlated with the proportion of revertants recovered after death. A82, a presumed double mutant showing low reversion, showed no detectable lethality. The pathogenicity of this mutant could be detected by inhibition of weight gain, which was proportional to the titer of virus injected. A low-level persistence, independent of the titer injected, occurred up to 7 days after injection. This was followed by complete clearance. It is concluded that the virulence of Sindbis virus may be considerably altered by mutation, and that this is related to events occurring at the cellular level.
Novel iron uptake system specified by ColV plasmids: an important component in the virulence of invasive strains of Escherichia coli
Authors: Williams P H
Published: January 2020

The enhanced virulence of invasive strains of Escherichia coli carrying ColV plasmids was shown to be due to a novel plasmid-mediated iron uptake system. Possession of a ColV plasmid conferred strong selective advantage on the host bacterial strain in experimental infections unless excess iron was administered in the inoculum. Moreover, supplementation of defined minimal medium with transferrin to complex available iron caused marked limitation of the growth of plasmid-free strains but had no effect on strains carrying a ColV plasmid. The activity of an efficient iron uptake process was clearly shown by experiments with a mutant of E. coli deficient in enterochelin biosynthesis. Although the mutant was dependent on the presence of citrate in the growth medium to facilitate iron transport, colicinogenic derivatives did not require added citrate for growth. Radioactive iron was shown to be taken up rapidly by nongrowing cells of the plasmid-carrying strain. Furthermore, it was observed that repression of the synthesis of specific outer membrane proteins normally induced by conditions of iron deficit was maintained after a shift of the colicinogenic strains from a rich medium to a medium low in iron. The ColV plasmid-mediated iron uptake system was independent of the active iron transport mechanisms known in E. coli, but like them it required tonB activity as a source of energy.
Requirements for fatty acid synthesis and a chelation-sensitive step in the production of glucosyltransferase by Streptococcus mutans
Authors: Kuramitsu H K , Wondrack L
Published: January 2020

The antibiotic cerulenin differentially inhibited the production of glucosyltransferase activity by Streptococcus mutans GS5. Cerulenin preferentially inhibited [14C]acetate incorporation into cellular lipids but did not affect protein synthesis or ribonucleic acid synthesis in the same manner. No significant intracellular accumulation of glucosyltransferase activity could be demonstrated in cultures treated with cerulenin. On the other hand, another inhibitor of lipid synthesis, sodium chlorophenoxyisobutyrate, did not differentially inhibit glucosyltransferase expression. In addition, the role of a metal-requiring protease in the production of glucosyltransferase activity was suggested by the observation that the chelator quinacrine differentially inhibited the production of the enzyme.
Serum antibodies to herpes simplex virus type 1 during active oral herpes infection
Authors: Ratner J J , Smith K O
Published: January 2020

Subjects with oral herpes lesions at the time of serum sampling had higher-efficiency antibody (higher proportion of neutralizing antibody as determined by plaque reduction, compared with total antibody as detected by radioimmunoassay) to herpes simplex virus type 1 (HSV-1) than did subjects with no lesions at the time of serum sampling. These higher-efficiency sera also had higher antibody titers to structural components of herpes simplex virus type 1 than did the low-efficiency sera. Absorption of high- and low-efficiency sera with purified herpes simplex virus type 1 particles removed all neutralizing antibody but not all antibody detected by radioimmunoassay. High-efficiency serum was depleted of more antibody to particulate antigen that was the low-efficiency serum, indicating that the high-efficiency serum contained a higher proportion of antibody to the virus particle.
Experimental syphilis and serological examination for treponematosis in hares
Authors: Horvath I , Kemenes F , Molnar L , Szeky A , Racz I
Published: January 2020

Of 202 captive hares studied, many of which had lesions on their external genital organs or testicular atrophy or both, 27% had positive serological tests for syphilis although dark-field examination of extracts of atrophic testes was negative. A total of 12 hares that were nonreactive for the serological test for syphilis was inoculated with Treponema pallidum, 9 intratesticularly and 3 intradermally. Six of the animals inoculated intratesticularly exhibited orchitis after 7 days with an associated accumulation of treponemes. No chancres developed in the intracutaneously inoculated animals during a 27-day period of observation. These results provide additional evidence to support the contention that endemic treponematosis occurs in wild hares and suggest that hares are moderately resistant to experimental infection with T. pallidum.
Virulence of Streptococcus mutans: revertants of mutant C4
Authors: Ikeda T , Otake S , Hirasawa M , Williams K , Kiyoyono H , McGhee J R , Shiota T
Published: January 2020

Mutant C4, a poor plaque-forming mutant of Streptococcus mutans 6715-HSR, was employed to obtain isolates resembling the parent strain (a plaque former). Seventeen presumptive revertants, as identified by colonial morphology, were isolated from mutant C4 after enrichment cycles in a sucrose-glass beads medium. These isolates displayed properties which resembled the parent in ability to produce plaque, patterns of fermentation, and resistance to streptomycin. In a detailed study, five selected isolates were found to be similar to the parent type 6715-HSR with respect to content of the serotype antigen, sucrose- or dextran-induced cell aggregation, glucosyltransferase and adherence activities, and cariogenicity. Thus, in selection for revertants to parental colonial morphology, the pleiotropic changes in plaque formation, adherence, glycosyltransferase activity, and virulence demonstrated by C4 all concomitantly reverted to their parental phenotypes.
Detection of a fibroblast proliferation inhibitory factor from Capnocytophaga sputigena
Authors: Stevens R H , Sela M N , Shapira J , Hammond B F
Published: January 2020

The addition of a sonic extract of Capnocytophaga sputigena to the culture fluid to human fibroblasts resulted in an inhibition of cell proliferation. The inhibition was dose-related (200 micrograms/ml caused a 90% inhibition, and 1,000 micrograms/ml caused a complete cessation of growth). The growth inhibition was not due to alterations in culture medium, pH or ionic strength, or to the effects of the C. sputigena lipopolysaccharide.
Effect of cultural conditions on trimethylsilyl-sugar profiles of Streptococcus mutans
Authors: Drucker D B , Aluyi H S
Published: January 2020

The cellular carbohydrates of Streptococcus mutans NCTC 10832 were converted to methyl glycoside trimethylsilyl ethers and analyzed by gas-liquid chromatography. The resulting profiles revealed the presence of glycerol, xylose, rhamnose, galactose, glucose, N-acetylglucosamine, and N-acetylmuramic acid. The proportions of monosaccharides in the profile were found to be stable with respect to changing growth temperature or duration of growth. However, the type of medium used, and its carbohydrate content, did significantly alter the profile, whereas gaseous atmosphere exerted a lesser effect on carbohydrate composition. By controlling growth parameters, we obtained reproducible profiles. Whole cells are probably a satisfactory alternative to highly purified cell walls.
Absorption of ablastic activity from mouse serum by using a Trypanosoma musculi population rich in dividing forms
Authors: Brooks B O , Reed N D
Published: January 2020

An in vivo assay of ablastin activity was developed by passive transfer of ablastic serum into nude mice. Preparations of Trypanosoma musculi that contained 35 to 50% dividing forms removed ablastic activity from mouse serum, but absorption of serum with a parasite preparation consisting of less than 5% dividing forms did not appreciably alter ablastic activity. These data suggest that ablastin is an antibody that can be absorbed onto homologous trypanosomes.
Enzyme-linked immunosorbent assay for antibodies to Toxoplasma gondii polysaccharides in human toxoplasmosis
Authors: Mineo J R , Camargo M E , Ferreira A W
Published: January 2020

A polysaccharide fraction from Toxoplasma gondii was adsorbed to polystyrene plates, and the enzyme-linked immunosorbent assay was performed (poly-ELISA) with peroxidase-labeled anti-immunoglobulin G and anti-immunoglobulin M antibodies. A comparison was made with a T. gondii total protein extract ELISA (protein ELISA) in serum samples presenting different toxoplasmosis serological patterns, as indicated by a battery of tests for toxoplasmosis. Very low titers and negative results were seen for immunoglobulin G poly-ELISA both for serum samples corresponding to ancient or transitional-period infections (serological patterns II and III) and for samples of recent or acute toxoplasmosis (pattern I). On the contrary, immunoglobulin M poly-ELISA furnished high titer results for pattern I sera, and a very close agreement of titers was seen between immunoglobulin M protein ELISA and immunoglobulin M poly-ELISA. When the polysaccharide fraction was added to pattern I sera, a complete blocking of immunoglobulin M antibody reactivity resulted only for poly-ELISA. In the same way, the total protein extract could completely block only reactivity for protein ELISA. In both cases, a limited decrease in titers was observed for respective heterologous assays.
Restriction endonucleases in identification of a genome type of adenovirus 19 associated with keratoconjunctivitis
Authors: Wadell G , de Jong J C
Published: January 2020

Adenovirus type 19 (Ad19) was first associated with disease in 1973 when several outbreaks of keratoconjunctivitis were reported from Europe and North America. We have examined Ad19 isolates by deoxyribonucleic acid restriction with BamHI, BglI, and SmaI restriction endonucleases. All keratoconjunctivitis-associated Ad19 isolates were identical but different from the Ad19 prototype. The total number of resolved restriction fragments of the Ad19 prototype genome was 31, only 17 of which migrated as the restriction fragments of keratoconjunctivitis-associated genomes. We conclude that two different genome types of Ad19 exist, one of which has been responsible for the recent outbreaks of keratoconjunctivitis.
Bacteriostatic effect of serum: role of antibody to lipopolysaccharide
Authors: Fitzgerald S P , Rogers H J
Published: January 2020

Previous work has shown that antibody and transferrin, acting together, exert a bacteriostatic effect on certain pathogenic Escherichia coli. This effect may be due to the ability of the antibody to interfere with the release of the iron chelator, enterochelin, from the bacterial cell. Enterochelin is essential for the transport of iron from transferrin to the bacterial cell. The nature of the bacterial antigen against which the antibody is directed has now been determined by means of adsorption experiments. It was found that absorption of serum either with hear-killed cells of E. coli O111 or with Boivin antigen abolished the bacteriostatic effect. A monosaccharide, which proved to be colitose (3,6-dideoxy-L-galactose), was isolated after acetic acid hydrolysis of the Boivin antigen. Colitose is the terminal monosaccharide of the O-specific side chain of the lipopolysaccharide from E. coli O111. This monosaccharide abolished the bacteriostatic effect of both whole serum and mixtures of antibody and iron-binding proteins. When administered by the intraperitoneal route, it reduced the resistance of mice to subsequent infection with E. coli O111. This ability of colitose to interfere with antibacterial mechanisms is in accord with published immunochemical studies.
Sialidase-Enhanced Lectin-Like Mechanism for
Actinomyces viscosus
and
Actinomyces naeslundii
Hemagglutination
Authors: Ellen R. P. , Fillery E. D. , Chan K. H. , Grove D. A.
Published: January 2020

Laboratory strains representing six numerical taxonomy clusters and fresh isolates of human
Actinomyces viscosus
and
Actinomyces naeslundii
were studied by standard flocculation slide tests for the ability to hemagglutinate erythrocytes (RBC) from various animal species. Human AB and horse RBC were agglutinated more frequently and rapidly than others; guinea pig RBC were agglutinated by only a few strains. Human AB RBC were selected for studies of hemagglutination mechanisms. Treatment of RBC with clostridial neuraminidase (NTRBC) greatly enhanced hemagglutination for almost all strains. In hapten inhibition experiments in which various concentrations of sugars were used, β-galactosides were the most effective inhibitors of hemagglutination for both RBC and NTRBC; inhibition of NTRBC agglutination required higher concentrations. Soybean lectin agglutinated both RBC and NTRBC but not
Actinomyces
cells. NTRBC agglutinated at a 125-fold-lower concentration. Hemagglutination was sensitive to ethylenediaminetetraacetate for one strain tested. Hemagglutination reactions were reversible by addition of β-galactosides. The ability of
Actinomyces
strains to “prime” RBC for hemagglutination by removing sialic acid to expose more penultimate β-galactoside sites was studied by recycling
Actinomyces
-agglutinated RBC which were dispersed with a lactose solution and washed free of bacteria (primed RBC). Priming in this manner augmented subsequent hemagglutination by indicator
Actinomyces
strains and made the RBC more sensitive to agglutination by soybean lectin. The priming ability of
Actinomyces
strains generally correlated with the amount of sialic acid removed from primed RBC. Strains representing the numerical taxonomy clusters differed in both their hemagglutinating and priming activities. Cluster 5 strains (typical
A. naeslundii
) were good agglutinators of RBC, NTRBC, and primed RBC but were poor primers. Cluster 3 strains (atypical
A. naeslundii
) were the weakest hemagglutinators but could prime RBC adequately for subsequent agglutination by other strains. Together, these data indicate that
Actinomyces
hemagglutination proceeds via a two-step mechanism: (i) neuraminidase removal of terminal sialic acid and (ii) lectin-like binding to exposed β-galactoside-associated sites on the RBC. Strains differ in the extent to which they can perform the two functions, and this specificity may relate to their taxonomic classification.
Shift in the prevalent human rotavirus detected by ribonucleic acid segment differences
Authors: Espejo R T , Muñóz O , Serafin F , Romero P
Published: January 2020

Rotavirus was purified from nine patients hospitalized with acute gastroenteritis from October to December, 1978, in Mexico City. Analysis of their ribonucleic acids by gel electrophoresis showed the presence of two distinct patterns (2s and 22) which had been observed in 1977, but which now were found in a very different proportion: the pattern called 2s, observed in only 11% (6 of 52) of the patients in 1977, was found in 90% (8 of 9) of the patients in 1978. Improvements in the electrophoretic method allowed us to observe differences in the migration of up to seven segments between the two patterns and to distinguish small differences in one or two segments within either of the two ribonucleic acid patterns.
Effects of cyclophosphamide on murine candidiasis
Authors: Moser S A , Domer J E
Published: January 2020

Male CBA/J mice were given a single dose of 200 mg of cyclophosphamide (CY) per kg 3 days before a first or second cutaneous inoculation with viable Candida albicans in an attempt to suppress antibody formation and determine the effects of such suppression on the development of acquired immunity. After cutaneous inoculation, mice not treated with CY developed acquired immunity to intravenous challenge, which was accompanied by the development of circulating antibodies, delayed hypersensitivity, and in vitro responsiveness of lymph node cells to Candida antigens. CY treatment resulted in an immediate depression of peripheral blood leukocytes, with polymorphonuclear leukocytes and monocytes rebounding quickly to normal or above normal levels while lymphocyte remained depressed throughout the 4-week observation period. In vitro stimulation of lymph node cells from CY-treated mice was depressed shortly after treatment; however, responses to phytohemagglutinin and three Candida antigens (a cell wall preparation, a membrane preparation, and soluble cytoplasmic substances) recovered, whereas the responses to lipopolysaccharide did not. CY effects on the cutaneous lesion were twofold; first, the number of viable Candida cells in the lesions was much higher in animals receiving CY 3 days before Candida inoculation, and second, the size of the dermal lesion was either greatly enhanced or reduced depending upon the time of CY treatment relative to the number of cutaneous Candida inoculations. CY-treated animals developed higher levels of delayed hypersensitivity to the membrane preparation when infected once cutaneously than did corresponding untreated animals. The number of mice responding with circulating antibodies to soluble cytoplasmic substances after cutaneous inoculation was greatly reduced in CY-treated groups, and this impaired ability to produce antibodies correlated with the poor survival of these mice after intravenous challenge. Our results suggest that the ability to produce antibody at the time of challenge is crucial to successful defense against systemic candidiasis in this murine model.
Biphasic protection against bacterial infection in mice induced by vaccination of Propionibacterium acnes
Authors: Kobayashi F , Nagoya T , Koshi T , Saino Y
Published: January 2020

A single intraperitoneal injection of the phenol-treated cells of Propionibacterium acnes into mice showed nonspecific resistance against subsequent lethal doses of an intraperitoneal challenge of Klebsiella pneumoniae, Staphylococcus aureus, and Streptococcus pyogenes. The protection showed a biphasic pattern. The maximum protection, designated as the early phase protection, was seen in mice injected with P. acnes vaccine 1 to 2 days before the challenge, whereas the late phase protection was seen in mice vaccinated 16 to 22 days before the challenge. The activity of the reticuloendothelial system in mice after vaccination also showed a biphasic pattern with the peak on days 4 and 12. The delayed activation of the reticuloendothelial system lasted up to 3 weeks and coincided with the period of the late phase protection. The early phase resistance was markedly impaired by the treatment with hydrocortisone and carrageenan, but not by the treatment with anti-thymocyte serum, actinomycin D, or cyclophosphamide. The number of peritoneal polymorphonuclear leukocytes in vaccinated mice increased on days 1 to 2. The number of macrophages also increased at 2 to 21 days after vaccination and reached its maximum on day 14. Total activities of acid phosphatase, Nitro Blue Tetrazolium reduction, and the phagocytic activities of peritoneal exudate cells were also enhanced on and after day 1 after the injection of P. acnes vaccine.
Comparison of two enzyme-linked immunosorbent assays for antigen quantitation: direct competition and antibody inhibition
Authors: Rissing J P , Buxton T B , Talledo R A , Sprinkle T J
Published: January 2020

Two enzyme-linked immunosorbent assays for antigen quantitation, direct competition and antibody inhibition, were used to measure rabbit immunoglobulin G in polystyrene microtiter plates and were compared for sensitivity and reproducibility. Both procedures repetitively detected this antigen in the 1- to 100-ng/ml range. Both procedures were predictably reproducible, with direct competition procedures having steeper slopes in ranges tested. Antibody inhibition does not require conjugated antigen and can detect sample antigens if available stock antigens can be passively bound to a solid-phase polystyrene plate.
Immunoglobulin G and M antibodies to pneumococcal polysaccharides detected by enzyme-linked immunosorbent assay
Authors: Barrett D J , Ammann A J , Stenmark S , Wara D W
Published: January 2020

An enzyme-linked immunosorbent assay has been developed to detect serum immunoglobulin G and M antibodies against pneumococcal polysaccharide antigens. Parameters affecting the specificity and sensitivity of the assay are described. A vigorous antibody response involving both the immunoglobulin G and M classes was demonstrated after pneumococcal polysaccharide immunization in normal adults. Studies with this enzyme-linked immunosorbent assay technique will allow further understanding of the biology of the primary and secondary immune response to Streptococcus pneumoniae in normals as well as in those persons most susceptible to infection with that organism.
Immunogenicity of ribosomes from enzymatically lysed Streptococcus pyogenes
Authors: Green B A , Johnson W
Published: January 2020

Ribosomal fractions isolated from Streptococcus pyogenes by physical and enzymatic disruption of the cell wall were found to provide protection in mice against challenge with the homologous M type. Although ribosomal fractions isolated by physical disruption of the cells also provided protection against challenge with several heterologous M types, ribosomal fractions from enzymatically lysed cells did not provide protection against any of the heterologous M types. Ribosomes isolated by either method were found to be contaminated with cell surface proteins. Chemical analysis of the ribosomes showed a greater protein:ribonucleic acid ratio in ribosomes from physically disrupted cells than in ribosomes from enzymatically disrupted cells (2:1 versus 1:1). Antisera to ribosomes isolated from physically disrupted cells detected many more antigenic determinants on ribosomes isolated from enzymatically disrupted cells than did the corresponding homologous antisera. Immunodiffusion analysis suggested that ribosomes isolated from physically disrupted cells may contain cell wall antigenic determinants which are present on ribosomes isolated from enzymatically disrupted cells in a partially degraded form. Washing of ribosomes in high-molarity salt solutions suggested that some of the contaminating cell wall proteins are tightly bound to the ribosomes.
Growth characteristics of group A streptococci in a new chemically defined medium
Authors: van de Rijn I , Kessler R E
Published: January 2020

A new chemically defined medium for the growth of group A streptococci has been formulated. The advantages of this new medium over previously described defined media are: (i) rates of growth (i.e., doubling times) of 20 strains were comparable to the rates of growth in complex media; (ii) each strain grew to a higher culture density in the new defined medium than in complex media; (iii) transfer from complex media with small inocula was possible without any prior adaptation regimen; and (iv) the production of virulence factors (i.e., M protein and hyaluronic acid) and extracellular enzymes during growth in this new medium was comparable to that in complex media.
Thioglycolate medium decreases resistance to bacterial infection in mice
Authors: Baker L A , Campbell P A
Published: January 2020

Brewer thioglycolate medium, a bacterial culture medium which is widely used as a nonspecific in vivo macrophage stimulant, was tested for its effect on mouse resistance to bacterial infection. Mice which did or did not receive thioglycolate medium were challenged with Listeria monocytogenes. Injection of thioglycolate medium significantly decreased the ability of the host to resist infection. This decreased resistance occurred whether thioglycolate medium was injected 0 to 9 days before, or 1 to 2 h after Listeria challenge. Bacteria grew considerably better in washed adherent peritoneal cells from thioglycolate medium-injected mice than in washed adherent peritoneal cells from normal mice. Thus, thioglycolate medium may ultimately cause a decrease in macrophage function.
Bacterial translocation from the gastrointestinal tract of athymic (nu/nu) mice
Authors: Owens W E , Berg R D
Published: January 2020

The immune system may be one host defense mechanism preventing viable indigenous bacteria from translocating from the mouse gastrointestinal lumen to the mesenteric lymph nodes, spleen, liver, or kidney. The role of T-cell-dependent immunity in preventing bacteria from translocating from the gastrointestinal tract was tested with congenitally athymic nude (nu/nu) mice, heterozygous (nu/+) mice, and thymus-grafted nude (nu/nu) mice. Viable bacteria were cultured from 50% of the mesenteric lymph nodes, spleens, livers, and kidneys of athymic (nu/nu) mice, whereas heterozygous (nu/+) mice exhibited viable bacteria in only 5.2% of these organs. Both aerobic and strictly anaerobic bacteria were cultured from these organs with Escherichia coli and Lactobacillus predominanting. Grafting thymuses to the athymic (nu/nu) mice restored their immunological responses to sheep erythrocyte antigens. The incidence of bacterial translocation from the gastrointestinal tract was reduced from 50% in the athymic (nu/nu) mice to 7.8% in the thymus-grafted (nu/nu) mice. Thus, T-cell-dependent immunity restored by thymic grafts inhibited the translocation of certain indigenous bacteria from the gastrointestinal tract to the spleen, liver, and kidney in nu/nu mice.
High-molecular-weight antigenic protein complex in the outer membrane of Neisseria gonorrhoeae
Authors: Newhall W J , Wilde C E , Sawyer W D , Haak R A
Published: January 2020

The outer membrane of Neisseria gonorrhoeae contains approximately 15 proteins, with 2 or 3 accounting for over 75% of the total protein mass. Samples of outer membrane from strain 2686 T4 analyzed by electrophoresis in 2% polyacrylamide gels revealed a band with an apparent molecular weight of 800,000. The band was protein material, as indicated by trypsin and pronase sensitivity and by L-[3H]proline incorporation. Peptidoglycan, nucleic acids, and carbohydrate were not detected in the band. Dye binding, L-[3H]proline incorporation, and labeling of solubilized outer-membrane proteins with 125I-labeled Bolton-Hunter reagent indicated that the band made up 10 to 13% of the total protein mass of isolated outer membranes. The material in the band was purified by gel filtration and, after reduction and alkylation, quantitatively recovered as subunits with an apparent molecular weight of 76,000. The protein in complex form was exposed at the cell surface, as evidenced by labeling whole cells with 125I by using a lactoperoxidase-catalyzed reaction and with CNBr-activated dextran. Rabbit serum raised against whole 2686 T4 gonococci contained antibody which reacted with the protein complex. The protein complex was detected in all gonococcal strains tested, but its presence could not be demonstrated in several other gram-negative species.
Enzyme-Linked Immunosorbent Assay for Quantitating the Humoral Immune Response to the Colonization Factor Antigen of Enterotoxigenic
Escherichia coli
Authors: Clegg Steven , Evans Dolores G. , Evans Doyle J.
Published: January 2020

The enzyme-linked immunosorbent assay technique was used to quantitate, in milligrams per milliliter, anti-colonization factor antigen/I (CFA/I) immunoglobulin G (IgG) in acute- and convalescent-phase sera of individuals who experienced diarrhea associated with CFA/I-positive enterotoxigenic
Escherichia coli
. Purified CFA/I was used as antigen to coat polystyrene Microtiter plate wells for the determination of anti-CFA/I antibody. A reference anti-CFA/I IgG preparation was obtained by affinity chromatography of a high-titered serum with a CFA/I-Sepharose 4B column; IgG was the only class of immunoglobulin detectable in this serum as anti-CFA/I. Goat anti-human IgG conjugated to alkaline phosphatase was used in the enzyme-linked immunosorbent assay. Quantitation of IgG in the reference anti-CFA/I serum was achieved by comparison with a known sample of pure human IgG. Anti-CFA/I in test sera was quantitated by titration with CFA/I-coated Microtiter plate wells in the enzyme-linked immunosorbent assay, using a standard curve obtained with the reference anti-CFA/I serum. Anti-CFA/I IgG in paired sera was determined as percentage of total IgG by using the radial immunodiffusion technique to quantitate total IgG for each test serum. Diarrhea with isolation of CFA/I-positive enterotoxigenic
E. coli
was associated with a significant rise in serum anti-CFA/I IgG when these values were expressed as either milligrams of IgG per milliliter or as percentage of total IgG, although the response varied quantitatively and nonresponders were detected. None of the matched controls showed an anti-CFA/I IgG response. Further elucidation of the immune response to enterotoxigenic
E. coli
can now be accomplished by applying these methods to determine the class and specificity of immunoglobulins in external secretions such as saliva and intestinal contents.
New Method for isolation of immunologically pure pili from Escherichia coli
Authors: Korhonen T K , Nurmiaho E L , Ranta H , Edén C S
Published: January 2020

A new technique for purification of bacterial pili was developed and applied to Escherichia coli strains isolated from the urine of patients with symptomatic urinary tract infections. After mechanical detachment from the bacterial cells, the pili were concentrated by precipitation with ammonium sulfate, dialyzed, and solubilized in buffer containing deoxycholate. The fraction containing the pili was purufied further by ultracentrifugation in a sucrose gradient and by elution through a Sepharose 4B column in 6 M urea buffer. The pilus filaments were not dissociated by concentrated urea and were eluted in the void volume of the column. The purified pili had a molecular weight of 17,000. The isoelectric point of the pili from one of the strains was 4.9, and about 43% of the amino acids were hydrophobic. Hyperimmunesera raised in rabbits against the purified pili did not contain detectable antibodies to the lipopolysaccharide O antigen or to the capsular polysaccharide K antigen of the homologous strain. The pili obtained by this purification procedure are free from other detectable bacterial surface antigens, and the purified pilus filaments are of relatively homogeneous size. This procedure enables purification of the pili also from flagellated strains.
Immunochemical differences between oral and nonoral strains of Bacteroides asaccharolyticus
Authors: Mansheim B J , Coleman S E
Published: January 2020

We undertook a morphological and immunochemical comparison of purified outer membrane antigens from oral and nonoral isolates of Bacteroides asaccharolyticus. Electron micrographs of thin sections of whole bacteria revealed a compact, electron-dense capsule external to the outer membrane of oral strains. A loose, web-like material was noted on the surface of several nonoral strains that was distinct from the dense capsule seen on oral strains. Polyacrylamide gel electrophoresis showed distinct differences in the protein band pattern between oral and nonoral isolates; sugar composition was similar with a few exceptions. An indirect fluorescent-antibody test utilizing antiserum to a purified capsular antigen from a single oral strain cross-reacted with all of numerous oral and nonoral strains of B. asaccharolyticus, thereby demonstrating a shared antigen that is species specific for B. asaccharolyticus. However, antibodies to an oral strain-derived capsular antigen were detectable by enzyme immunoassay only in serum from rabbits immunized with oral strains. Thus, definite morphological and immunochemical differences were found between oral and nonoral isolates of B.asaccharolyticus.
Characterization of a flagellar sheath protein of Vibrio cholerae
Authors: Hranitzky K W , Mulholland A , Larson A D , Eubanks E R , Hart L T
Published: January 2020

A flagellar sheath protein of Vibrio cholerae CA401 (Inaba) was characterized. Purity of the preparation was indicated by a single band on polyacrylamide gel electrophoresis gels and on Ouchterlony plates prepared with antibody against crude sheath material. The sheath protein was composed of three polypeptides with minimal molecular weights of 61,500, 60,000, and 56,500. The presence of sheath protein on the flagellum as well as on the outer membrane of the cell was demonstrated by ferritin labeling experiments with antiserum. Sheath protein antibody reacted similarly in labeling experiments and agglutination tests with a classical Ogawa strain and two nonagglutinating V. cholerae isolates, indicating that the sheath protein may represent the common Vibrio H antigen. Antibody specific for lipopolysaccharide labeled the cell but not the sheathed flagellum, which demonstrated that the sheath is not a simple extension of the outer membrane of the cell.
Binding of streptococcal antigens to muscle tissue in vitro
Authors: Stinson M W , Nisengard R J , Bergey E J
Published: January 2020

Antigens extracted from cells of Streptococcus pyogenes T6 and Streptococcus mutans strains AHT, BHT, 10449, OMZ175, and K1R adsorbed to the sarcolemmal sheath of cardiac muscle cells in vitro. Similar preparations from S. salivarius, S. sanguis, Staphylococcus aureus, and Lactobacillus casei had weak or negligible tissue-binding activity. Tissue-bound bacterial antigens were detected with homologous rabbit antisera with both indirect immunofluorescence tests and an indirect radioimmunoassay. Serological cross-reactivity was observed between the tissue-binding factors of S. pyogenes and S. mutans cells but not between the bacteria and muscle tissue. In a comparative study of extraction procedures, the greatest yield of tissue-binding factors was obtained from group A streptococci by cell disruption in buffer at 4 degrees C. Hot aqueous phenol and hot water extracts were inactive. Antibodies specific for the tissue-binding factor(s) were readily adsorbed from rabbit anti-S. pyogenes serum by a preparation of isolated cytoplasmic membranes but not by a suspension of cell wall fragments. The heart-binding component of S. pyogenes cell extracts was inactivated by protease digestion and heat treatment and to a lesser extent by periodic acid oxidation. The capacity of heart cell components to adsorb streptococcal antigens was reduced by protease treatment but not by the action of neuraminidase, hyaluronidase, organic solvents, or detergents.
Adherence of Pseudomonas aeruginosa to tracheal cells injured by influenza infection or by endotracheal intubation
Authors: Ramphal R , Small P M , Shands J W , Fischlschweiger W , Small P A
Published: January 2020

Adherence of Pseudomonas aeruginosa to normal, injured, and regenerating tracheal mucosa was examined by scanning electron microscopy. Uninfected and influenza-infected murine tracheas were exposed to six strains of P. aeruginosa isolated from human sources and one strain of platn origin. All of the strains tested adhered to desquamating cells of the infected tracheas, but not to normal mucosa, the basal cell layer, or the regenerating epithelium. Adherence increased when the incubation time of the bacteria with the trachea was prolonged. Strains isolated from human tracheas appeared to adhere better than strains derived from the urinary tract. After endotracheal intubation of ferrets, P. aeruginosa adhered only to the injured cells and to areas of exposed basement membrane. We call this phenomenon "opportunistic adherence" and propose that alteration of the cell surfaces or cell injury facilitates the adherence of this bacterium and that adherence to injured cells may be a key to the pathogenesis of opportunistic Pseudomonas infections.
Investigation of the component of Propionibacterium acnes (Corynebacterium parvum) responsible for macrophage activation
Authors: Buck G E , Kelly M T
Published: January 2020

Mouse peritoneal macrophages, activated with Propionibacterium acnes (Corynebacterium parvum) in vivo, exhibited altered morphological characteristics, increased acid phosphatase activity, and release of [EH]thymidine-labeled DNA from tumor cells in vitro. Comparison with macrophages from mice injected with a control organism, P. jensenii, showed that the morphological changes, but not acid phosphatase, correlated with the development of tumoricidal activity. Investigation of the microbial component responsible for this activity indicated it was heat stable and refractory to extraction by methanol-chloroform, dilute acid, butanol, or a serial combination of extraction procedures. When the cells were disrupted by mechanical breakage, no activity could be found in the cell wall, soluble cytoplasm, or particulate cytoplasm. These results suggest that intact organisms are required for macrophage activation, and they may resolve conflicting reports on the nature of the immunostimulating activity of P. acnes.