bluuebunny/crossref_metadata_2025 huggingface dataset:
Influence of Antibiotics or Certain Intestinal Bacteria on Orally Administered
Candida albicans
in Germ-Free and Conventional Mice
Authors: Clark J. Derrell
Published: January 2020

Candida albicans
, administered by gastric intubation, persisted in the gastrointestinal tract of gnotobiotic mice for long periods but was eliminated within a relatively short period of time in pathogen-free mice. Oxytetracycline administered by mouth had no reproducible effect on the persistence of
C. albicans
in the gastrointestinal tract of either germ-free or pathogen-free mice. Prolonged administration of streptomycin extended the time that
C. albicans
could be recovered from feces of pathogen-free mice when compared to mice not receiving the antibiotic or those receiving a single large dose. There was a brief interval of time during which
C. albicans
could not be recovered from the feces of gnotobiotic mice contaminated with certain intestinal bacteria, but eventually all mice began to shed the fungus again.
C. albicans
administered by mouth was not pathogenic for germ-free or pathogen-free mice. It can be concluded from these findings that mice do not possess an innate resistance to
C. albicans
but that pathogen-free mice do possess some ecological mechanism which prevents establishment of the fungus in their gastrointestinal tract. The reason for the difference in colonization of
C. albicans
in germ-free or gnotobiotic mice and pathogen-free mice was not determined.
Effect of Diuresis on
Staphylococcus aureus
Kidney Infections in Mice
Authors: Furtado Dolores
Published: January 2020

In the Cornett strain of mice, water diuresis did not prevent hematogenous production of pyelonephritis by
Staphylococcus aureus
. Increased fluid intake did not affect the numbers of organisms deposited in the kidneys or the rate of growth during the first 4 hr after inoculation. Drinking the glucose solution did not enhance bacterial proliferation within the renal parenchyma. Subcutaneous injection of saline to supplement for interruption of drinking after inoculation reduced the numbers of organisms recovered in the kidneys but not sufficiently to prevent production of pyelonephritis. Incorporating penicillin as a marker indicated that fluids administered by subcutaneous injections were rapidly delivered to the kidneys. Combining diuresis with treatment did not influence the rapidity of delivery of antimicrobial to the kidneys or the length of time that it was present in the renal homogenate.
Isolation and possible relevance of Thermoactinomyces candidus proteinases in farmer's lung disease
Authors: Roberts R C , Nelles L P , Treuhaft M W , Marx J J
Published: January 2020

The thermophilic actinomycetes are the most common etiological agents causing hypersensitivity pneumonitis. Antigen preparations of these organisms contain proteolytic activity. Further investigation of the proteinases of the thermophilic actinomycetes was undertaken to determine whether this activity may contribute directly to the pathogenesis of hypersensitivity pneumonitis and pulmonary mycotoxicosis. The presence of proteolytic activity in aerosolized dust from moldy silage was demonstrated, and antibodies to two proteolytic enzymes from Thermoactinomyces candidus were found in the blood of farmer's lung patients who had been sensitized to this organism. These two enzymes were isolated from culture filtrate antigen preparations that had been partially characterized with respect to the proteolytic activities and their interaction with human serum proteinase inhibitors. Both proteinases belonged to the serine class of endopeptidases. Neither proteinase was inhibited by alpha 1-proteinase inhibitor or alpha 1-antichymotrypsin. Both proteinases were inhibited by alpha 2-macroglobulin. One of the proteinases had elastase activity. Inhalation of these proteinases apparently does occur, and they may induce an inflammatory response in the lungs since they are not inhibited by the main proteinase inhibitors protecting the lung.
Specificities of monoclonal antibodies against the activated delta-endotoxin of Bacillus thuringiensis var. thuringiensis
Authors: Huber-Lukac M , Lüthy P , Braun D G
Published: January 2020

Eight hybrid cell lines secreting monoclonal antibodies directed against the activated delta-endotoxin of Bacillus thuringiensis var. thuringiensis were grown in BALB/c mice. Ascites fluids were collected, and the antibodies were purified by antigen-affinity chromatography. The specificity of each monoclonal antibody for the toxin and protoxin was established by the indirect enzyme-linked immunosorbent assay. All the antibodies consisted of gamma 1 heavy and kappa light chains. They were reactive with both the native toxin and the protoxin. In contrast to specific goat antiserum, they failed, however, to bind to heat and sodium dodecyl sulfate denatured antigen. These eight cloned cell lines gave rise to five kinds of antibodies distinguished by isoelectric focusing. Competitive antibody binding studies revealed that these five antibodies recognize at least four distinct antigenic determinants of the native toxin and the protoxin. Two of the epitopes are unrelated, whereas three antibodies compete for binding to their antigenic determinants. In the bioassay with larvae of Pieris brassicae, one antibody was found to block the toxin and protoxin activity completely. A second inhibited it partially, whereas the other three antibodies did not affect it at all.
Purification and characterization of a Shigella dysenteriae 1-like toxin produced by Escherichia coli
Authors: O'Brien A D , LaVeck G D
Published: January 2020

A toxin from an enteropathogenic strain of Escherichia coli (E. coli H30) was purified to apparent homogeneity from cell lysates. The steps used to isolate the E. coli H30 toxin included French pressure-cell disruption of bacteria grown in iron-depleted media. Affi-Gel Blue chromatography, chromatofocusing, and anti-Shiga toxin affinity chromatography. The mobilities of the subunits of radioiodinated E. coli H30 toxin and Shiga toxin observed after the two toxins were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis were identical. In the absence of 2-mercaptoethanol, a narrow band was seen at Mr 31,500 (+/- 1,000), and a wide heavy band was observed between Mr 4,000 and 15,000. In the presence of 2-mercaptoethanol, bands were seen at Mr 31,500 (+/- 1,000), 27,000, and 4,000 to 15,000. Other similarities between purified E. coli H30 and Shiga 60R toxins included identical isoelectric points (7.03 +/- 0.02); comparable biological activities, i.e., cytotoxicity, lethality for mice, and enterotoxicity; and the same relative heat stabilities (up to 65 degrees C for 30 min). Nevertheless, the two toxins had apparently different molecular weights as determined by sucrose gradient analysis, by gel filtration, and by cross-linking experiments with dimethyl suberimidate. The Mr of native E. coli H30 toxin estimated from cross-linking studies was 48,000, whereas the estimated Mr of Shiga 60R toxin was 58,000. These results suggest that like the cholera-E. coli-heat-labile toxin family, a family of Shiga-like toxins exists.
Neutrophil circulation and release from bone marrow during hypothermia
Authors: Biggar W D , Bohn D , Kent G
Published: January 2020

The effect of hypothermia on neutrophil circulation and release from bone marrow has been studied. Pigs were anesthetized and maintained at 37 degrees C or surface cooled to 29 degrees C over 60 min. As the core temperature was reduced to 29 degrees C, the number of circulating neutrophils (X 10(9) per liter) fell from 6.0 +/- 0.6 to 2.3 +/- 0.3 by 60 min. No significant change in the number of circulating mature or immature neutrophils was observed over the 4 h of observation at 29 degrees C. Neutrophil demargination after administration of intravenous catecholamines was similar at 37 and 29 degrees C. Steroid stimulation of bone marrow to release neutrophils was markedly impaired at 29 degrees C. Circulating mature neutrophils in normothermic pigs increased from 5.6 +/- 1.2 to 10.4 +/- 1.2 by 120 min after administration of intravenous hydrocortisone sodium succinate. Circulating immature neutrophils increased from 1.7 +/- 0.3 to 5.3 +/- 0.4. At 29 degrees C, no significant changes in the number of circulating mature or immature neutrophils occurred. Endotoxin also failed to stimulate neutrophil release from the bone marrow. Furthermore, a marked neutropenia occurred in hypothermic pigs after intravenous endotoxin, which persisted for the 3 h of observation. Neutrophil circulation and release from bone marrow are compromised by hypothermia, which may increase the risk for bacterial infection.
Attachment of
Pneumocystis carinii
to Type I Alveolar Cells Studied by Freeze-Fracture Electron Microscopy
Authors: Yoneda Kokichi , Walzer Peter D.
Published: January 2020

Pneumocystis carinii
was tightly attached to host alveolar type I cells, as judged by freeze-fracture electron microscopy. In contrast to other organisms studied by this technique, no changes in the cell membranes of
P. carinii
or the host cells could be demonstrated. These data suggest that
P. carinii
attaches in an unusual manner.
Do lymphocytes from Chagasic patients respond to heart antigens?
Authors: Todd C W , Todd N R , Guimaraes A C
Published: January 2020

Lymphocyte transformation studies of nonadherent lymphocytes from chronic Chagasic and uninfected persons demonstrated that responses of all individuals to a mouse heart homogenate showed a correlation with responses to streptococcal antigens. Considering the known cross-reactions between streptococcal and cardiac antigens and the high reactivity of Chagasic patients to streptococcal antigens, it is possible that positive lymphocyte transformation to unfractionated heart antigen preparations may not represent specific reactivity to heart antigens.
Cellular Immunity in a Cutaneous Model of Cryptococcosis
Authors: Domer Judith E. , Lyon Frank L. , Murphy Juneann W.
Published: January 2020

The temporal development of cellular immune responses in mice inoculated cutaneously with viable
Cryptococcus neoformans
145 was determined in vivo and in vitro by comparing several antigen preparations for their efficacy in the assays selected. Three antigens derived from
C. neoformans
145, viz., a culture filtrate preparation (CneF-145), a membrane extract (B-HEX), and soluble cytoplasmic substances (SCS), were compared for their ability to detect delayed hypersensitivity (DH) in vivo in a footpad assay or to stimulate lymphocytes in vitro in a thymidine incorporation assay. DH to B-HEX could be demonstrated as early as 1 week after infection, whereas significant responses to SCS and CneF-145 were not regularly detected until 3 weeks after infection. Substantial reactions were observed to all three antigens up to 12 weeks, although they peaked at 2 to 3 weeks. Reactions to B-HEX and SCS were somewhat better than those to CneF. Differences in the efficacies of the three antigens were not obvious after the sixth week of infection, however. In vitro, lymph node cells from infected animals were stimulated significantly with all three antigens beginning at week 1. As with DH, however, responses to CneF-145 were usually less than those to SCS and B-HEX. In vitro lymphocyte responses waned after approximately 6 weeks, whereas DH responses were clearly positive through 12 weeks. In addition to the studies in infected animals, animals immunized with heat-killed cells of
C. neoformans
145 or 184 were tested 6 to 8 days later for DH with CneF-145, CneF-184, or B-HEX derived from
C. neoformans
145. The CneF-145 and CneF-184 were equally effective for detecting DH, regardless of the cryptococcal strain used for immunization. Likewise, the B-HEX detected equivalent responses in mice sensitized with each cryptococcal strain. Since all three antigens were soluble and easily extracted and since each elicited significant cellular immune responses in infected animals, further studies involving their specificity and the nature of their reactive components seems warranted as they may help evaluate immune responses in humans infected with this fungus.
Plasmids that code for production of colonization factor antigen II and enterotoxin production in strains of Escherichia coli
Authors: Smith H R , Scotland S M , Rowe B
Published: January 2020

Similar plasmids that code for production of colonization factor antigen II and enterotoxins were mobilized into Escherichia coli K-12 from strains of E. coli O6.H16 biotype B or C that expressed colonization factor antigen II components CS2 and CS3. Further transfer experiments demonstrated that there was production of CS2 and CS3 in an O6.H16 biotype C strain, but there was expression of only CS3 in other serogroups and in E. coli K-12.
Interaction of viral envelope glycoproteins with fibronectin
Authors: Julkunen I , Hautanen A , Keski-Oja J
Published: January 2020

An interaction between fibronectin and viral envelope glycoprotein micelles isolated from influenza A, parainfluenza 1, and mumps viruses was found by enzyme immunoassay. All three different glycoprotein micelles bound efficiently to solid-phase fibronectin. When fibronectin was permitted to bind to solid-phase viral glycoproteins, dose-dependent binding was observed. Soluble glycoprotein micelles inhibited the binding of fibronectin to immobilized glycoprotein preparations. The binding was not observed when fibronectin was pretreated with neuraminidase, suggesting that the sugar moieties of fibronectin are responsible for the affinity. This affinity may play a role in virus-cell interactions or in the opsonization of certain viruses during infection.
Role of granulocytes and monocytes in experimental Staphylococcus epidermidis endocarditis
Authors: Meddens M J , Thompson J , Bauer W C , Hermans J , van Furth R
Published: January 2020

The role of granulocytes and monocytes during the induction and course of Staphylococcus epidermidis endocarditis was investigated by the selective depletion of monocytes with the drug VP16-213 and of both granulocytes and monocytes with nitrogen mustard. The induction of endocarditis was influenced only by the depletion of monocytes: the 50% infective dose differed significantly, being 3.4 X 10(5) CFU in control rabbits and 3.4 X 10(4) CFU in the monocyte-depleted rabbits, whereas no significant differences were found between the latter and those depleted of both granulocytes and monocytes. Also, control rabbits injected with 10(6) or 10(7) CFU had a significantly higher incidence of sterile vegetations than did rabbits selectively depleted of granulocytes or monocytes. Compared with baseline values, mean monocyte numbers at the time of bacterial inoculation were significantly increased in control rabbits whose vegetations remained sterile, whereas this effect was not seen in rabbits whose vegetations became infected. The course of the endocarditis appeared to be significantly influenced by both granulocytes and monocytes. Comparison showed that a decrease of the same numbers of these cells per microliter of blood was accompanied for the monocytes by an approximately fourfold higher increase of the number of staphylococci in the vegetations. The correlation between the number of granulocytes and of monocytes on the one hand and the number of staphylococci in the vegetations on the other was not substantially influenced by the duration of the disease or the number of staphylococci injected to induce the endocarditis. The number injected proved to be significantly correlated with the number of staphylococci in the vegetations. In rabbits with numbers of CFU per gram of vegetation exceeding 10(7), blood cultures were usually positive. This finding applied rarely to control rabbits, but generally to drug-treated rabbits. In the latter animals a significant correlation between the number of staphylococci in the vegetations and in the circulation was found. We conclude that only monocytes have a measurable effect on the induction of Staphylococcus epidermidis endocarditis but during its course both granulocytes and monocytes keep the endocardial infection in check.
Cooperation between antibodies and monocytes that inhibit in vitro proliferation of Plasmodium falciparum
Authors: Khusmith S , Druilhe P
Published: January 2020

The cooperative effect between nonsensitized monocytes from normal individuals and malarial antibody in depressing parasite multiplication was investigated in an in vitro assay. The addition of purified normal monocytes to Plasmodium falciparum cultures in the presence of serum from immune individuals markedly inhibited the proliferation of the parasite in vitro: the parasitemia observed was about 22 times lower than that in the presence of immune serum alone. This cooperative effect was found to be effective over a wide range of monocyte/erythrocyte ratios (1:10 to 1:160) and serum concentrations (1/10 to 1/80). Immunoglobulin G extracted on protein A-Sepharose was as effective as total serum in this system. These data suggest that cooperation between nonsensitized cells and immunoglobulins could be an important effector mechanism against P. falciparum parasites in vivo.
Separation and purification of the hemagglutinins from Bordetella pertussis
Authors: Sato Y , Cowell J L , Sato H , Burstyn D G , Manclark C R
Published: January 2020

The role of the filamentous hemagglutinin (FHA) and the lymphocytosis-promoting factor hemagglutinin (LPF) in pertussis pathogenesis and immunity is the subject of active investigation. To be certain of their role as protective antigens, the hemagglutinins must be pure and free of each other. This report describes procedures to separate and purify FHA and LPF from the culture supernatant of stationary cultures of Bordetella pertussis Tohama, using hydroxylapatite, haptoglobin-Sepharose, and Sepharose CL-6B filtration chromatography. Purified FHA contained less than 0.002% active LPF assayed by histamine-sensitizing activity, and both hemagglutinins contained less than 0.01% of each other based on antigenic activity measured by an enzyme-linked immunosorbent assay, using affinity chromatography-purified antibody to each hemagglutinin. LPF and FHA were also shown to be antigenically distinct by immunodiffusion and were judged to be highly purified proteins by polyacrylamide gel electrophoresis. In addition, the purification procedures yielded milligram amounts of each hemagglutinin with very good recovery of starting activities.
Rash produced by mouse cytomegalovirus
Authors: Chong K T , Mims C A
Published: January 2020

A striking rash was produced in mice 4 days after intravenous injection of large doses of murine cytomegalovirus. Routine histology showed cell infiltration, vascular damage, and hemorrhage. Immunofluourescent staining showed dermal but not epidermal infection, and this appeared to be initiated after the localization in small blood vessels of circulating virus or of virus-containing leukocytes. The rash was accompanied by a high level of complement-requiring neutralizing antibodies in the serum and extensive deposition of immune complexes in the renal glomeruli containing immunoglobulin G, immunoglobulin M, and complement. When mice were depleted of complement by cobra venom factor, when antibody responses were reduced by cyclophosphamide treatment, or when a prostaglandin inhibitor (indomethacin) was given, the rash was much less severe. The role of direct viral damage and inflammatory and immune responses in the genesis of the lesions is discussed.
Detection of antibodies to human coronaviruses 229E and OC43 in the sera of multiple sclerosis patients and normal subjects
Authors: Hovanec D L , Flanagan T D
Published: January 2020

Sera collected from 90 multiple sclerosis patients and 148 age-matched normal subjects were examined for the presence of antibodies against human coronaviruses (HCV) 229E and OC43 by enzyme immunoassay (EIA). The results demonstrated no significant difference between the MS patients and the normal subjects in their antibody titer to HCV 229E and HCV OC43. Further analysis of these 238 sera indicated that a stronger EIA reaction was generally observed against HCV OC43 (mean EIA value at an optical density of 492 nm = 0.896) than against HCV 229E (mean EIA value at an optical density of 492 nm = 0.346).
Prevalence of viridans streptococci exhibiting lactose-inhibitable coaggregation with oral actinomycetes
Authors: Kolenbrander P E , Williams B L
Published: January 2020

Fresh oral isolates from human dental plaque were selected on the basis of their spherical morphology. In a double-blind experiment, their species identity and ability to coaggregate with human oral Actinomyces viscosus and Actinomyces naeslundii were determined. Of the 110 isolates characterized, 30 were identified as either Streptococcus mutans, Streptococcus anginosus-constellatus, or Veillonella parvula; none of these coaggregated with the actinomycetes. The remaining 80 isolates were identified as Streptococcus sanguis, Streptococcus mitis, Streptococcus MG-intermedius, or Streptococcus morbillorum. Of these 49 isolates (61%) coaggregated with actinomycetes, and nearly 90% (43 of 49 isolates) exhibited lactose-inhibitable coaggregations. Compared with previously characterized coaggregation properties determined with stock culture strains of streptococci, 82% of the fresh isolates exhibited identical coaggregations. The other 18% made up a new coaggregation group that possessed a related lactose-inhibitable coaggregation pattern. Thus, most fresh isolates that coaggregated exhibited lactose-inhibitable coaggregations with human oral actinomycetes. It is suggested that these coaggregations are mediated by a network of lectin-carbohydrate interactions similar to those already characterized in previous studies with stock cultures of actinomycetes and streptococci.
Neuraminidase production by a Streptococcus sanguis strain associated with subacute bacterial endocarditis
Authors: Straus D C , Portnoy-Duran C
Published: January 2020

The properties of an extracellular neuraminidase produced by a Streptococcus sanguis strain (isolated from a confirmed case of subacute bacterial endocarditis) during growth in a defined medium was examined in this investigation. This enzyme, isolated from concentrated culture supernatants of S. sanguis biotype II, was active against human alpha-1 acid glycoprotein, N-acetylneuramin lactose, bovine submaxillary mucin, and fetuin. Neuraminidase production paralleled bacterial growth in defined medium and was maximal in the early stationary phase of growth but decreased dramatically, probably owing to protease production, during the late stationary phase. The enzyme was purified to near homogeneity by a combination of salt fractionation, ion-exchanged chromatography on DEAE-Sephacel, and gel filtration on Sephadex G-200. These procedures yielded an enzyme preparation that possessed a specific activity of 174.4 mumol of sialic acid released per min per mg of protein against human alpha-1 acid glycoprotein. The Km value for this enzyme with human alpha-1 acid glycoprotein as substrate was 2.5 X 10(-3) M, and the enzyme possessed a pH optimum of 6.5. The S. sanguis neuraminidase had a molecular weight of approximately 85,000 as estimated by gel filtration and approximately 90,000 when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme was stable at temperatures of 4 and 37 degrees C for 3 h, but approximately 50% of the enzymatic activity was lost within 30 min at 50 degrees C, with 100% of the enzymatic activity being destroyed within 10 min at temperatures of greater than or equal to 65 degrees C.
Lethal synergism induced in mice by influenza type A virus and type Ia group B streptococci
Authors: Jones W T , Menna J H , Wennerstrom D E
Published: January 2020

Intranasal inoculation of CD-1 or BALB/c mice with low doses of influenza A/PR8/34 (HON1) virus followed 48 h later by intranasal inoculation of low doses of type Ia group B streptococci effected a lethal synergism. At a constant input dose of virus, a direct relationship between input dose of bacteria and percent mortality was observed; the converse was also true. An inverse relationship between input dose of group B streptococci, but not input dose of virus, and mean time to death was observed in CD-1 but not in BALB/c mice. The kinetics of influenza A/PR8/34 virus and group B streptococcal replication in singly and dually infected BALB/c mice was determined by assaying samples from the lungs, liver, spleen, and blood for viable group B streptococci and infectious influenza A/PR8/34 virus. No significant difference in virus replication in the lung was observed between singly and dually infected mice. Extrapulmonary dissemination of virus was not observed. Concurrent virus infection effected a 10,000- to 100,000-fold increase in the levels of type Ia group B streptococci in the lung. Potentiation of group B streptococcal infection of the lung was not associated with bacteremia or infection of the liver or spleen, a finding contrary to previous observations of fulminant septicemia after intranasal inoculation of mice with input doses of group B streptococci less than one-tenth of the pulmonary levels observed in the present study.
Siderophore production by Vibrio vulnificus
Authors: Simpson L M , Oliver J D
Published: January 2020

Previous studies in our laboratory, as well as clinical evidence, have suggested that increased iron levels in the host may be important in infections caused by the halophilic pathogen Vibrio vulnificus. To study iron acquisition, we induced siderophore production by growth in a low-iron medium, and biochemical testing indicated the production of both hydroxamate- and phenolate-type siderophores. The siderophores were extracted from growth filtrates with ethyl acetate (for phenolates) and phenol-chloroform-ether (for hydroxamates). These extracts enhanced the growth of V. vulnificus when the bacterium was grown in iron-limited medium. The ability of these siderophores to stimulate the growth of Salmonella typhimurium LT-2 enb-7 (a mutant deficient in the biosynthesis of enterochelin) and Arthrobacter flavescens JG-9 (a hydroxamate auxotroph) supported the conclusion that V. vulnificus produces both hydroxamate- and phenolate-type siderophores.
A set of two monoclonal antibodies specific for the cell surface-exposed 39K major outer membrane protein of Haemophilus influenzae type b defines all strains of this pathogen
Authors: Gulig P A , Frisch C F , Hansen E J
Published: January 2020

Six murine plasma cell hybridomas producing monoclonal antibodies (mabs) directed against the 39,000-molecular-weight (39K) major outer membrane protein of Haemophilus influenzae type b were employed in the antigenic analysis of the 39K protein. The initial characterization of the mabs by radioimmunoprecipitation analysis showed that four of these mabs reacted with antigenic determinants of the 39K protein that are exposed on the bacterial cell surface and accessible to antibody. The other two mabs reacted with antigenic determinants of the 39K protein that are either not exposed on the H. influenzae type b cell surface or not accessible to antibody (internal determinants). A total of 126 clinical isolates of H. influenzae type b obtained from pediatric research centers throughout the United States were examined for reactivity with the six mabs by using a solid-phase radioimmunoassay in which bacterial colony growth from agar plates was placed on filter paper and used as antigen. The reactivities of these strains with two of the mabs recognizing cell surface-exposed antigenic determinants of the 39K protein were used to divide the 126 strains into four different groups. Group 1 strains reacted with mab 12D9, group 2 strains reacted with mab 4C4, group 3 strains reacted with both mabs 12D9 and 4C4, and group 4 strains (only one was found) did not react with either mab. The reactivities of two other mabs recognizing cell surface-exposed antigenic determinants of the 39K protein were used to further divide the four groups into eight subgroups. A single mab recognizing an internal antigenic determinant of the 39K protein reacted with every H. influenzae type b strain examined in this study. These data indicate that only limited antigenic heterogeneity exists among the cell surface-exposed antigenic determinants of the 39K outer membrane proteins among H. influenzae type b strains and that at least one internal antigenic determinant of the 39K protein is universally present in all H. influenzae type b strains. Radioimmunoprecipitation analysis also demonstrated that H. influenzae type b strains which lacked a 39K major outer membrane protein possessed a 38K major outer membrane protein which reacted with the anti-39K mabs, indicating that the 38K and 39K outer membrane proteins of different H. influenzae type b strains are antigenically related.
Chemical properties of heat-stable enterotoxins produced by enterotoxigenic Escherichia coli of different host origins
Authors: Dreyfus L A , Frantz J C , Robertson D C
Published: January 2020

Five heat-stable enterotoxins (STs) produced by enterotoxigenic Escherichia coli strains of porcine, bovine, and human host origins have been purified to apparent homogeneity. The STs with biological activity in suckling mice and piglets (STaS) contained 18 amino acid residues, 10 amino acids with a high proportion of acidic amino acids, and 6 half-cystines. The carboxy-terminal and amino-terminal residues of all STaS were tyrosine and asparagine, respectively. All five STa preparations were homogeneous by several criteria: (i) a single symmetrical peak on gel filtration, (ii) a single fluorescent band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, (iii) single carboxyl-terminal and amino-terminal residues, and (iv) amino acid analysis data that indicated a stoichiometric relationship between the component amino acids. The isoelectric points of the five STaS ranged from 3.88 to 4.08. All five purified preparations were heat stable and not denatured by organic solvents, detergents, or treatment at pH 1, but were partially inactivated after incubation at pH 12.0. Biological activity was completely abolished by treatment with reducing and oxidizing agents, which suggested that one or more disulfide bonds play an important role in the mechanism of action of STaS. Antisera raised against strain 431 STa, produced by a porcine class II enterpathogen, neutralized homologous 431 STa as well as heterologous purified STa preparations. The antiserum was used as a reagent in a sensitive radioimmunoassay to detect suckling mouse-positive strains of enterotoxigenic E. coli.
Interaction of a rat lung lectin with the exopolysaccharides of Pseudomonas aeruginosa
Authors: McArthur H A , Ceri H
Published: January 2020

The specific interaction between the exopolysaccharide purified from a number of Pseudomonas aeruginosa isolates from cystic fibrosis patients and a rat lung heparin-lectin was assayed. The polysaccharide prepared from Homma serotypes M, B, I, and G did not act as hapten inhibitors of lectin activity, whereas the polymers prepared from ca. 80% of strains that did not type with Homma serum did act as hapten inhibitors. Inhibition was shown not to be due to lipopolysaccharide. The infrared spectrums of both inhibitory and noninhibitory polymers appeared very similar, although small amounts of glucose and an unidentified amino sugar were found only in the nontypable strains. This evidence suggests that rat lung lectin recognizes and distinguishes a specific type of alginate-like polymer prevalent on the Homma nontypable P. aeruginosa.
Role of adherence in the pathogenesis of Haemophilus influenzae type b infection in infant rats
Authors: Kaplan S L , Mason E O , Wiedermann B L
Published: January 2020

We evaluated the role of pili in the pathogenesis of disease due to Haemophilus influenzae type b (HiTb), using the infant rat model. Piliated and nonpiliated HiTb strains were isolated from the nasopharynx and cerebrospinal fluid, respectively, of three children. Infant rats inoculated intranasally with nonadherent HiTb developed bacteremia and meningitis more frequently (P = 0.005) than animals inoculated with companion adherent HiTb strains. When analyzed separately, only one HiTb pair (884/880) demonstrated significant differences in the incidence of bacteremia and meningitis between the adherent and nonadherent strains. Blood or cerebrospinal isolates recovered from infant rats inoculated with piliated adherent HiTb strains were not piliated and were not adherent in vitro. Adherent and nonadherent HiTb colonized the nasopharynx of infant rats equally. The piliated strains of HiTb were not adherent in vivo or in vitro to rat nasal or buccal epithelial cells, respectively. Piliated strains of HiTb have no apparent advantage over nonpiliated HiTb strains for colonization or invasion of infant rats. Furthermore, the loss of piliation is noted for cerebrospinal fluid, blood, and nasal isolates of HiTb cultured from infant rats inoculated with an adherent piliated HiTb strain. Thus, the loss or suppression of pili may be an important prerequisite for the invasion of the host by HiTb strains that are highly piliated.
Induction of interleukin 1 secretion by adjuvant-active peptidoglycans
Authors: Vacheron F , Guenounou M , Nauciel C
Published: January 2020

The ability of differently structured, purified peptidoglycans (PG) to induce interleukin 1 (IL1) secretion was compared. PG from Bacillus megaterium and Staphylococcus aureus stimulated the production of IL1 by mouse peritoneal macrophages and human adherent mononuclear cells, whereas PG from Micrococcus lysodeikticus and Corynebacterium poinsettiae were inactive. There was a correlation between the ability of PG to induce IL1 secretion and previously demonstrated immunoenhancing activities (adjuvant effect, increase of resistance to tumor growth) of PG. PG solubilization by lysozyme decreased but did not abolish the PG effect on IL1 secretion. Active PG induced IL1 production in nude mice and in the C3H/HeJ strain (which is unresponsive to lipopolysaccharides).
Production of superoxide anion, prostaglandins, and hydroxyeicosatetraenoic acids by macrophages from hypersensitivity-type (Schistosoma mansoni egg) and foreign body-type granulomas
Authors: Chensue S W , Kunkel S L , Higashi G I , Ward P A , Boros D L
Published: January 2020

Macrophages isolated from hypersensitivity (Schistosoma mansoni egg) and foreign body- (Sephadex bead) type granulomas were evaluated with regard to superoxide anion (O2-) production and arachidonic acid metabolism. Granuloma macrophages from schistosome-infected mice were examined during both the acute and modulated phases of the disease. In addition, the populations were characterized phenotypically by measurement of Ia antigen expression. Based on differences in the parameters studied at least three different macrophage populations could be identified in acute, modulated, and foreign body-type lesions, respectively. Macrophages from acute lesions (8-week granuloma macrophages) produced significant amounts of O2-, prostaglandins, and monohydroxyeicosatetraenoic acids without the addition of an exogenous stimulus. These cells also showed a high degree of Ia expression. In contrast, macrophages from modulated (20-week granuloma macrophages) and foreign body (foreign body granuloma macrophages) lesions required stimulation with phorbol ester to evoke significant O2- production and minimally metabolized arachidonic acid. However, 20-week and foreign body granuloma macrophages could be distinguished by their high and low degrees of Ia expression, respectively. The role of lymphokines and other intercellular signals in determining macrophage activation states within granulomas is discussed.
Characterization of Spiroplasma mirum (suckling mouse cataract agent) in a rabbit lens cell culture
Authors: Megraud F , Gamon L B , McGarrity G J
Published: January 2020

Spiroplasma mirum (suckling mouse cataract agent) was studied in an epithelial cell line AG-4676, derived from rabbit eye lens. Rabbit eye lens is a natural target tissue of S. mirum infection. The organism grew rapidly in this cell line, reaching titers of 10(7) to 10(9) color change units per ml at 7 days after infection. This is the same level as that achieved in SP-4 medium designed specifically for S. mirum. No lag period was apparent in growth in AG-4676. S. mirum did not grow in Dulbecco minimal essential medium-10% fetal bovine serum, the medium for AG-4676, indicating the need for cells or a cellular product. S. mirum-infected AG-4676 cells exhibited vacuolization and granulation and an increase in polynucleation compared with uninfected controls (36/100 versus 14/100, P less than 0.001). Infection significantly decreased the growth rate of AG-4676, especially late in the growth cycle. In a representative experiment, growth of AG-4676 at 11 days was reduced from 9 X 10(5) to 2 X 10(4) cells by S. mirum infection. S. mirum grew to high titers in conditioned medium of AG-4676, obtained from cell-free supernatants of 1- to 5-day-old AG-4676 cultures. This growth promotion was not due to osmotic conditioning of the medium. Preliminary characterization of this growth promotion substance showed it to be active after 0.22-micron filtration, heating at 56 degrees C for 30 min, freezing and thawing, and dilution at 10(-1) but not 10(-2). AG-4676-propagated S. mirum produced death or cataracts in suckling Wistar rats at the same frequency (55/60, 91.7%) as SP-4-propagated organisms (60/65, 92.3%).
Bacterial endotoxin isolated from a water spray air humidification system as a putative agent of occupation-related lung disease
Authors: Flaherty D K , Deck F H , Cooper J , Bishop K , Winzenburger P A , Smith L R , Bynum L , Witmer W B
Published: January 2020

Outbreaks of hypersensitivity pneumonitis or humidifier fever were attributed to the inhalation of organic material aerosolized by a chilled-water spray humidification system. The purpose of this study was to isolate and characterize the serologically detectable antigen(s) present in extracts obtained from the humidification system. By using bicarbonate or phenol-water extractions or both, the antigen was isolated and characterized, using colorimetry, gas-liquid chromatography, reverse-phase high-performance liquid chromatography, and X-ray fluorescence. Carbohydrates, hexosamines, phosphorus, and even-numbered saturated and unsaturated fatty acids were constituents of the serologically detectable antigen. When tested in in vivo and in vitro assays, the antigen had demonstrable endotoxin activity. All subjects with biopsy-proven lung disease and a majority of subjects suspected of having lung disease had antibodies directed toward the purified endotoxin. The data strongly suggest that an aerosolized bacterial endotoxin is a putative agent inducing lung disease.
Effect of ammonia on in vivo and in vitro immune responses
Authors: Targowski S P , Klucinski W , Babiker S , Nonnecke B J
Published: January 2020

The effects of exposure of animals to ammonia on their delayed type of dermal response, the mitogenic and antigenic responses of their lymphocytes, and the bactericidal and phagocytic activities of their alveolar macrophages were examined. Experimental guinea pigs vaccinated with Mycobacterium bovis BCG were exposed to 3.75 micrograms of ammonia per dl of air (50 ppm) or 6.75 micrograms of ammonia per dl of air (90 ppm), whereas control animals also vaccinated with BCG were maintained in the normal environment. The delayed type of dermal response to tuberculin injected 3 weeks later was significantly (P less than 0.05) less in experimental animals exposed to 6.75 micrograms of ammonia per dl than in control animals. In vitro, the response of blood lymphocytes and bronchial lymphocytes to phytohemagglutinin, concanavalin A, and tuberculin stimulation was significantly (P less than 0.01) less than the response of lymphocytes from control animals. The response of normal blood lymphocytes to phytohemagglutinin incubated in medium containing 1 or 10 mg of ammonia per dl was significantly (P less than 0.01) reduced as compared with the response of lymphocytes incubated without ammonia. The viability of lymphocytes incubated with these concentrations of ammonia was significantly (P less than 0.01) affected. There was no significant difference in the bactericidal or phagocytic activities of alveolar macrophages collected from animals exposed to ammonia and control animals. However, ammonia added to the culture of alveolar macrophages from normal animals significantly inhibited their bactericidal activity.
Monoclonal antibodies distinguish phase variants of Coxiella burnetii
Authors: Williams J C , Johnston M R , Peacock M G , Thomas L A , Stewart S , Portis J L
Published: January 2020

Monoclonal antibodies (MAbs) directed against phase I and II variants of Coxiella burnetii were produced by fusing myeloma SP2/O-AG 14 cells with spleen cells from mice immunized with the chloroform-methanol extraction residue of phase I whole cells. Two hybridoma clones which distinguished the phase variants by microimmunofluorescence assay were isolated and characterized. The MAbs showing specificity for phase I cells (MAbI-1, immunoglobulin G, subclass 3 kappa) reacted with the hot phenol-water extract of phase I C. burnetii in immunodiffusion and enzyme-linked immunosorbent assays. MAbI-1 reacted with high-molecular-weight components from phase I phenol-water extract and whole cell in an immunoblot assay. Specificity of MAbI-1 for a carbohydrate epitope in the phenol-water extract was demonstrated by periodic acid inactivation of binding by a competitive enzyme-linked immunosorbent assay. Phase I antigenic sites were apparently well represented on the surface of cells as demonstrated by complete fluorescence and microagglutination. The MAb showing specificity for phase II cells (MAbII-1, immunoglobulin G, subclass 2b kappa) reacted with whole cells in the microimmunofluorescence assay, microagglutination test, complement fixation test, and the enzyme-linked immunosorbent assay. MAbII-1 reacted specifically with a 29,500-dalton surface protein as demonstrated by immunoprecipitation of 125I-surface-labeled cells. Although MAbII-1 reacted with detergent-solubilized protein, it did not react with sodium-dodecyl sulfate-denatured protein by immunoblot assay. This protein was not exposed on the surface of phase I cells, but chloroform-methanol extraction of phase I cells exposed the phase II epitope.
Effect of iron on antibacterial immunity in vaccinated mice
Authors: Kochan I , Wagner S K , Wasynczuk J
Published: January 2020

The effect of iron on resistance to Salmonella typhimurium was investigated in mice inoculated with vaccines prepared from live and avirulent (SL3770) or killed and virulent (SR11 or LT2) bacteria. It has been found that mice vaccinated with SL3770 vaccine develop an immunity which can be neutralized with iron. Iron promoted the development of lethal infections by serving as a growth-essential nutrilite for infecting bacteria and by neutralizing the acquired immunity. The titration of this dual effect of iron showed that more iron was needed to neutralize the immunity in vaccinated animals than to promote bacterial growth in normal animals. In the presence of a sufficient amount of exogenous iron, as few as 10 bacteria caused lethal infections in normal and immune mice with the same effectiveness. This iron-sensitive immunity could be changed to iron-resistant immunity by the immunological stimulation of SL3770-vaccinated mice with a sonicated vaccine prepared from heat-killed SR11 or LT2 bacteria. In distinction to iron-sensitive immunity, iron-resistant immunity could be transferred from SR11- or LT2-stimulated to normal mice with serum. Although effective in the transfer of antibacterial immunity, sera of SR11- or LT2-stimulated mice supported the growth of virulent bacteria as well as did sera of normal mice. The absorption of immune serum with either SR11 or LT2 bacteria removed its protective quality, but the sensitized bacteria remained as infectious as untreated bacteria for iron-treated normal mice. Only in SL3770-vaccinated mice were the immune serum-sensitized bacteria not able to cause the infection in spite of daily treatment with iron. These results suggest that iron-resistant immunity is due to the synergistic action of specific antibody and phagocytes of immunologically stimulated animals.
Lipopolysaccharide interaction with rabbit erythrocyte membranes
Authors: Carr C , Morrison D C
Published: January 2020

In this study we have characterized the association of a polysaccharide-deficient, lipid-rich lipopolysaccharide (LPS) with rabbit erythrocytes (RaRBC). With polymyxin B sulfate-mediated hemolysis as a probe, we have shown that Salmonella minnesota R595 LPS interacts with RaRBC in two distinguishable steps. The first step whereby RaRBC exposed to LPS are rendered sensitive to polymyxin B-initiated lysis probably represents absorption of LPS to the RaRBC membrane. We investigated two possible mechanisms for the subsequent time-dependent decrease in response of LPS-treated RaRBC to polymyxin B. We found that the decay in polymyxin B susceptibility of LPS-treated RaRBC cannot be attributed to a decrease in binding of LPS to the RaRBC. On the other hand, our results are consistent with a time-dependent rearrangement of the amphipathic LPS within the lipid bilayer of the RaRBC membrane. In particular, at lower incubation temperatures of RaRBC and LPS, the decay in polymyxin B-induced hemolysis is slower, presumably, because the increased membrane viscosity allows less rapid rearrangement of LPS within the lipid bilayer. A putative hydrophobic intercalation of LPS into a mammalian cell membrane may be of importance in LPS stimulation of responsive cells.
Structural and antigenic heterogeneity of lipopolysaccharides of Campylobacter jejuni and Campylobacter coli
Authors: Logan S M , Trust T J
Published: January 2020

The techniques of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, silver staining, and immunoblotting were used to analyze the lipopolysaccharide (LPS) structure of 20 strains of Campylobacter jejuni and 4 strains of Campylobacter coli belonging to more than 22 thermostable serotypes. The LPSs of all strains examined were shown to be of a low-molecular-weight type, and these low-molecular-weight LPSs conferred heat-stable serospecificity. High-molecular-weight banding observed with both in vivo LPS in proteinase K digests of whole cell lysates and purified LPS was shown to be due to the ready ability of Campylobacter lipopolysaccharide to form aggregates rather than to the presence of O polysaccharide chains. Purified LPSs from two strains of C. jejuni were also subjected to gross chemical analysis. The high-lipid A to low-neutral sugar ratio of both LPSs was typical of LPSs lacking O polysaccharide chains.