bluuebunny/crossref_metadata_2025 huggingface dataset:
Infection of gnotobiotic pigs with an Escherichia coli O157:H7 strain associated with an outbreak of hemorrhagic colitis
Authors: Francis D H , Collins J E , Duimstra J R
Published: January 2020

Gnotobiotic pigs inoculated with an Escherichia coli O157:H7 strain isolated from a human with hemorrhagic colitis developed anorexia, lethargy, and watery diarrhea. Bacteria diffusely colonized the cecum and colon surfaces and the crypt epithelium. At bacterial attachment sites, microvilli were effaced, and epithelial cells were irregularly shaped, rounded, or detached. Submucosa, lamina propria, and mesentery were markedly edematous and contained many inflammatory cells.
Detection of anti-Vibrio vulnificus cytolysin antibodies in sera from mice and a human surviving V. vulnificus disease
Authors: Gray L D , Kreger A S
Published: January 2020

An enzyme-linked immunosorbent assay and a cytolysin neutralization assay were used detect anti-Vibrio vulnificus cytolysin antibodies in sera from mice and a human that survived V. vulnificus disease. The detection of antibodies against the cytolysin indicated that the cytolysin is produced in vivo, and this observation is consistent with the hypothesis that the cytolysin is involved in the pathogenesis of V. vulnificus disease.
Nucleotide sequence of the type A streptococcal exotoxin (erythrogenic toxin) gene from Streptococcus pyogenes bacteriophage T12
Authors: Weeks C R , Ferretti J J
Published: January 2020

The gene specifying type A streptococcal exotoxin (speA), also known as erythrogenic toxin, was cloned from the Streptococcus pyogenes bacteriophage T12 genome and analyzed by nucleotide sequencing. The speA gene consists of 753 base pairs and codes for a 29,244-molecular-weight protein. The speA gene product contains a putative 30-amino acid signal peptide, resulting in a molecular weight of 25,787 for the secreted protein. A possible promoter and ribosome-binding site are present in the region upstream from the speA gene, and a transcriptional terminator is located 69 bases downstream from the translational termination codon. The amino acid sequence of the carboxy-terminal portion of the type A streptococcal exotoxin exhibits extensive homology with the carboxy terminus of Staphylococcus aureus enterotoxins B and C1.
Increased susceptibility to lethal Candida infections in burned mice preinfected with Pseudomonas aeruginosa or pretreated with proteolytic enzymes
Authors: Neely A N , Law E J , Holder I A
Published: January 2020

Lethal Candida infections in burn patients are frequently preceded by or occur concomitantly with bacterial infections, which are often due to Pseudomonas aeruginosa. In this study, we developed a burned, mixed-challenge mouse model, which was designed to determine whether and how a recent bacterial infection could influence the development of subsequent candidosis. In this model, burned mice that were preinfected with a sublethal challenge of elastase-producing P. aeruginosa strain WR-5 and then sublethally challenged with Candida albicans exhibited a mortality rate of 60%, while unburned mice challenged in the same way and burned mice that received only one challenge organism exhibited mortality rates of less than 10%. Quantitative microbial counts performed with the kidneys, livers, and eschars of burned mice challenged with both organisms indicated that the deaths were due to Candida infection. Substitution of an elastase-negative P. aeruginosa strain for strain WR-5 in the model resulted in significantly lower mortality rates and lower microbial numbers in the organs. When the Pseudomonas enzyme elastase was substituted for the elastase-positive bacteria in the model, both the mortality rates and the organ counts were comparable to the values found after preinfection with strain WR-5. Another protease, thermolysin, was substituted for the elastase and produced similar mortality results. When the protease inhibitor alpha 2-macroglobulin was given to burned mice infected with the two organisms, it prevented the deaths due to Candida infection. We concluded that this model is one way to study bacterial-fungal infections in burned mice, that recent Pseudomonas infections could predispose burned mice to fatal candidosis, and that the proteolytic activity generated by the bacteria was primarily responsible for the establishment of lethal fungal infections.
Infant mouse model of adherence and colonization of intestinal tissues by enterotoxigenic strains of Escherichia coli isolated from humans
Authors: Goldhar J , Zilberberg A , Ofek I
Published: January 2020

The ability of enterotoxigenic Escherichia coli H10407, which possesses colonization factor antigen I, to colonize the intestinal mucosa of infant mice was considerably better than that of its colonization factor antigen I-negative derivative H10407-P. The latter strain previously was shown to lack cell adhering ability in vitro and to have a diminished capacity to infect human volunteers as compared with the parent strain. D-Mannose blocked both colonization by an enterotoxigenic E. coli isolate (801) possessing both mannose-resistant and mannose-sensitive adhesins and the in vitro adherence of the strain to intestinal segments of infant mice. A derivative of another enterotoxigenic E. coli strain (lacking both mannose-sensitive and mannose-resistant adhesins obtained by in vivo passage showed a significant increase in colonizing ability in comparison with the parent strain. We conclude that the infant mouse model of infection of intestinal mucosa complemented by in vitro adherence assays with excised intestinal tissue is suitable for the study of the bacterial properties responsible for the various stages of intestinal colonization by human enterotoxigenic E. coli.
Surface antigen detected by a Schistosoma mansoni monoclonal antibody in worm extracts and kidney deposits of infected mice and hamsters
Authors: Carneiro C R , Lopes J D
Published: January 2020

Four monoclonal antibodies (MAbs) derived from a Schistosoma mansoni-infected mouse reacted against the tegument or the cell layer of the digestive tract of the adult worm. They also showed similar patterns of immunofluorescence staining when schistosomula were used as antigens. Two of the MAbs (4A10 and 4D3) recognized immune complexes deposited in the kidneys of infected mice and hamsters as detected by indirect and direct immunofluorescence reactions. When adsorbed to polystyrene beads, both MAbs allowed the quantitative detection of antigen by an enzyme immunoassay. 125I-labeled 4A10 binding to live schistosomula and corresponding inhibition assay ruled out the possibility that this binding could be through its Fc fragment. The same MAb detected an antigen migrating as an 80-kilodalton protein by Western blot analysis of soluble worm antigen preparation after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Use of MAb 4A10 may help to elucidate mechanisms of renal pathology and also be of value in the development of immunodiagnostic assays.
Evidence for inhibition of fusion of lysosomal and prelysosomal compartments with phagosomes in macrophages infected with pathogenic Mycobacterium avium
Authors: Frehel C , de Chastellier C , Lang T , Rastogi N
Published: January 2020

Bone marrow-derived cultured macrophages were infected with the pathogenic organism Mycobacterium avium. Immediately after infection and at 1 to 28 days later, cells either were stained for acid phosphatase activity or given horseradish peroxidase, which served as a pinocytotic marker. With the former, fusions between phagosomes and lysosomes exclusively were assessed; with the latter, those between phagosomes and both pinosomes and lysosomes were determined. As a control, similar experiments were undertaken by infecting macrophages with gamma ray-killed M. avium and the nonpathogenic live organisms Mycobacterium aurum and Bacillus subtilis. After infection with live M. avium, fusions between phagosomes and acid phosphatase-positive vesicles (lysosomes) were inhibited. The same inhibition was observed whether phagosomes contained damaged or structurally intact (presumed to be live) bacteria, except for the early time points. This inhibition was, however, partial, suggesting that some of the live bacteria are resistant to the hydrolytic enzymes of the phagolysosomal environment. Fusions between horseradish peroxidase-positive vesicles (pinosomes and lysosomes) and phagosomes depended upon the morphological state of the bacteria. Damaged bacteria did not inhibit fusions, whereas with intact bacteria, a partial inhibition which increased with time was observed. The two types of experiment suggest that viable M. avium can impair phagosome-pinosome fusions.
Preferential inhibition of primary granule release from bovine neutrophils by a Brucella abortus extract
Authors: Bertram T A , Canning P C , Roth J A
Published: January 2020

A low-molecular-weight Brucella abortus extract (a nucleotidelike material) inhibited zymosan-elicited neutrophil degranulation and trichloroacetic acid-precipitable protein iodination (a measure of myeloperoxidase and H2O2 release from neutrophilic leukocytes). Inhibition of neutrophil function was directly related to the concentration of the Brucella extract. The extract preferentially inhibited degranulation of primary (azurophilic or peroxidase positive) granules and had limited inhibition of secondary (specific or peroxidase negative) granule release but did not inhibit opsonized zymosan ingestion. Inhibition of protein iodination closely paralleled that of primary granule release but was unrelated to inhibition of secondary granule release. These results suggest that B. abortus has a component which is capable of inhibiting release of myeloperoxidase by dose-dependent preferential inhibition of primary granule release from bovine neutrophilic leukocytes.
Purification and characterization of Vibrio cholerae non-O1 heat-stable enterotoxin
Authors: Arita M , Takeda T , Honda T , Miwatani T
Published: January 2020

A toxin which causes rapid fluid accumulation in a suckling mouse assay and which was produced by Vibrio cholerae non-O1 was investigated. The toxin was purified from the culture supernatant of V. cholerae non-O1 (strain A-5) by ammonium sulfate fractionation, hydroxyapatite treatment, ethanol extraction, column chromatographies on SP-Sephadex C-50 and DEAE-Sephadex A-25, and high-pressure liquid chromatography on a Lichrosorb RP-8 column. About 1.4 X 10(5)-fold purification was achieved, with a recovery of about 12%. Although the crude preparation was heat labile, the purified toxin was heat stable. The minimum effective dose of purified toxin was about 5 ng in the suckling mouse assay. The amino acid composition of the purified toxin was determined to be Asp(3), Glu(1), Ala(1), half-Cys(6), Ile(2), Leu(1), Phe(1), and Pro(1). These data show the production of a new type of heat-stable enterotoxin (NAG-ST) by V. cholerae non-O1.
Pathophysiology of experimental leishmaniasis: pattern of development of metastatic disease in the susceptible host
Authors: Hill J O
Published: January 2020

A clear understanding of the etiology of the various forms of leishmaniasis will require knowledge of how physiological properties of the parasite and host immunity influence the pattern of development of the disease. Of particular importance are how these factors affect the growth rate of Leishmania spp. at the site of inoculation in the skin, their capacity to disseminate to visceral and distant cutaneous sites, and their capacity to multiply once there. This paper details the pattern of development of disseminated Leishmania major infection in susceptible BALB/c nu/+ and BALB/c nu/nu mice. It was found that the parasite disseminates from the hind footpad to distant cutaneous sites soon after metastatic foci are established in the liver and spleen. Both mononuclear phagocytes and neutrophils may be the vehicles for the transport of the parasite in the blood. Once visceral and cutaneous metastases are established, the parasites in those foci increase in number progressively. L. major has the capacity to multiply at visceral and cutaneous sites at the same rate. Despite the presence of viable parasites in a number of skin sites, cutaneous metastatic lesions developed almost exclusively on the feet and the tail. Furthermore, these lesions appeared to develop preferentially at sites near joints, suggesting that factors other than temperature may influence the development of cutaneous metastatic lesions.
Polyclonal activation of rat splenic lymphocytes after in vivo administration of Mycoplasma pulmonis and its relation to in vitro response
Authors: Williamson J S , Davis J K , Cassell G H
Published: January 2020

The plaque-forming cell (PFC) assay with sheep erythrocytes (SRBC) sensitized with different antigens and a 4-h tritiated thymidine pulse assay were used to determine whether polyclonal activation occurs in rats following in vivo administration of Mycoplasma pulmonis. Injection of M. pulmonis into F344 rats resulted in an increase in the number of splenic immunoglobulin M-secreting PFC that produced antibodies reactive with the trinitrophenyl hapten and with SRBC. This polyclonal response reached a peak by 72 h after injection and returned to normal levels by 96 h, at which time the specific response to M. pulmonis reached its peak. Heat treatment and preopsonization of M. pulmonis with antiserum before injection resulted in reduced numbers of PFC against M. pulmonis-sensitized SRBC, trinitrophenyl hapten-sensitized SRBC, and SRBC. The number of PFC against the three types of target cells also increased in LEW rats after immunization with M. pulmonis. The number of PFC against SRBC and staphylococcal protein A-sensitized SRBC was higher in immunized LEW rats than in immunized F344 rats. Examination of unimmunized animals also revealed that LEW rats had higher initial numbers of PFC than did F344 rats. These results showed that polyclonal activation occurs in rats following in vivo administration of M. pulmonis and that LEW rats have an inherent propensity to develop higher nonspecific responses in vivo than F344 rats.
Double-blind analysis of the relation between adult periodontitis and systemic host response to suspected periodontal pathogens
Authors: Gmür R , Hrodek K , Saxer U P , Guggenheim B
Published: January 2020

By using a sensitive enzyme-linked immunosorbent assay, 200 randomly selected sera from Red Cross blood donors were screened for immunoglobulin G (IgG), IgA, and IgM levels against Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, and Bacteroides intermedius. A subgroup of 79 blood donors was clinically examined for type and extent of periodontal destruction, and serological and clinical data were subjected in all possible dual combinations to correlation analyses. The results revealed that the majority of the blood donors suffered from moderate to severe adult periodontitis, often coupled with severe gingival inflammation. No cases of localized juvenile periodontitis or rapidly progressive periodontitis were observed. The extent of periodontal destruction proved to be significantly correlated only to the IgG response levels against B. gingivalis. Corresponding correlation tests assessing the relationships of loss of attachment, bone loss, pocket depth, and papillary bleeding index with the IgG responses to A. actinomycetemcomitans were of marginal significance, while the IgG responses to B. intermedius revealed no relationship to the periodontal health status. The specific IgM responses proved to be unrelated to the clinical parameters, but interestingly, they were found to be highly correlated with each other. Specific IgA levels were frequently too low for enzyme-linked immunosorbent assay testing and, therefore, had to be exempted from statistical analyses. Assessments of the serotype specificity of strongly elevated IgG responses to A. actinomycetemcomitans disclosed no evidence for an association of a particular serotype-specific IgG response with the occurrence of adult periodontal destruction. In contrast to results of earlier studies, a number of sera were found to contain strongly elevated IgG levels against two or even all three serotypes. Although derived by an alternative approach, the reported results largely corroborate earlier observations linking only the occurrence of elevated anti-B. gingivalis IgG responses to the presence of marked periodontal lesions in adults.
Identification of suppressive components in "Haemophilus somnus" fractions which inhibit bovine polymorphonuclear leukocyte function
Authors: Chiang Y W , Kaeberle M L , Roth J A
Published: January 2020

"Haemophilus somnus" fractions which inhibited iodination of protein by bovine polymorphonuclear leukocytes were isolated by heat extracting a washed bacterial suspension at 60 degrees C or incubating the bacterial suspension at 37 degrees C and were partially purified by ultrafiltration. The components in each fraction were separated by reverse-phase high-performance liquid chromatography and identified as ribonucleotides, a ribonucleoside, and purine and pyrimidine bases. Most of the compounds were found to be inhibitory to iodination in a dose-dependent manner. When the effect of each component on iodination at the concentrations present in "H. somnus" fractions was determined, it was found that guanine and GMP were the components responsible for most of the suppression in the fraction isolated by heat extraction, whereas guanine and adenine were the major inhibitory components in the fraction isolated by incubation at 37 degrees C.
Appearance of Fc receptors on polymorphonuclear leukocytes after migration and their role in phagocytosis
Authors: Targowski S P , Niemialtowski M
Published: January 2020

The effect of the migration of bovine blood polymorphonuclear leukocytes (PMNs) in vitro on their phagocytic activity was studied. PMNs were examined before and after migration through various membranes for rosette formation with sensitized sheep erythrocytes to detect Fc receptors (FcRs), phagocytic activity mediated through FcRs with opsonized staphylococci (Smith strain), and phagocytic activity mediated through nonimmunological receptors with unopsonized staphylococci (strain 305). Migration of PMNs was observed from the upper to the lower compartment of the blind well chamber through Millipore and Nuclepore membranes; through Millipore, Nuclepore, and nylon membranes coated with collagen; and through collagen-coated Millipore, Nuclepore, and nylon membranes overlaid with MA-104, BHK-21, MDBK-99, TB, or FBHE cells. Random migration of PMNs toward the plain medium (the same medium in the upper and lower compartments) through the membranes with and without a monolayer of cells increased the percentage of PMNs forming rosettes. In contrast, migration toward the medium containing lipopolysaccharide (LPS), formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP), or zymosan-activated serum (Act. serum) did not change the percentage of PMNs forming rosettes. The increased percentage of PMNs forming rosettes was associated with the enhanced phagocytosis of opsonized staphylococci (mediated by FcRs). In contrast, migration of PMNs toward LPS, FMLP, or Act. serum did not enhance phagocytosis mediated through FcRs. However, PMNs after migration toward LPS, FMLP, Act. serum, and plain medium enhanced phagocytosis of unopsonized staphylococci (mediated through the nonimmunological receptors).
Immunoglobulin G (IgG) and IgA subclass pattern of human antibodies to Shigella flexneri and Salmonella serogroup B and D lipopolysaccharide O antigens
Authors: Persson M A , Ekwall E , Hammarström L , Lindberg A A , Smith C I
Published: January 2020

The subclass distribution of human serum antibodies to the O-antigenic lipopolysaccharides of Salmonella serogroups B and D and to Shigella flexneri serotypes 1b, 2a, and 4a lipopolysaccharide antigens were analyzed in an enzyme-linked immunosorbent assay with monoclonal antibodies to the immunoglobulin subclasses. The patients had culture-verified Salmonella (17 Swedes) or Shigella flexneri (23 Vietnamese; 11 children and 12 adults) infections. Consecutive samples drawn during 1 year postinfection were investigated. Antibodies to the Salmonella antigens were mainly of the immunoglobulin G1 (IgG1), IgA1, and IgA2 subclasses. For the Salmonella serogroup B O polysaccharide, the IgA1 and IgA2 subclasses had peak values earlier than (6/9) or coinciding with the IgG1 (3/9) peak value. Furthermore, the IgA2 response to Salmonella serogroup B was positively correlated to the duration of the carrier state (P less than 0.001); the corresponding IgA1 response was less well correlated but was still significant (P less than 0.02). In the case of the Shigella flexneri O polysaccharide, specific antibodies appeared mainly in the IgG1 and IgA1 subclasses. Some IgG2 was also found, surprisingly even in very young patients. No subclass shift with time within the immunoglobulin classes was noted in any of the groups.
Inhibitory effect of substituted aromatic hydrocarbons on adherence of Escherichia coli to human epithelial cells
Authors: Falkowski W , Edwards M , Schaeffer A J
Published: January 2020

Adherence of Escherichia coli to epithelial cells is postulated to be a necessary step in the pathogenesis of urinary tract infections. Investigations have focused on the role of carbohydrates in adherence because of the ability of mannose to inhibit adherence; however, there have been several reports of an additional hydrophobic receptor. Here we describe the inhibition of adherence mediated by substituted aromatic hydrocarbon compounds and the relationship of our findings to support the role of a postulated hydrophobic-binding domain.
Lymphokine-activated-killer-mediated lysis of cells infected with typhus group rickettsiae can be inhibited by OKT3 monoclonal antibody
Authors: Carl M , Dasch G A
Published: January 2020

We examined the ability of monoclonal antibodies directed against lymphocyte surface antigens to block the lysis of typhus group rickettsia-infected cells by lymphokine-activated killer effectors. Lysis was significantly inhibited by OKT3 monoclonal antibody but not by OKT4 or OKT8 monoclonal antibody.
A reexamination of the O1 lipopolysaccharide antigen group of Escherichia coli
Authors: Moll A , Kusecek B , Pluschke G , Morelli G , Kamke M , Jann B , Jann K , Achtman M
Published: January 2020

A total of 64 Escherichia coli strains of the O1 serogroup were tested for the migration pattern of their lipopolysaccharides (LPS) on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. O1:K1 and O1:K51 strains of the OMP5 outer membrane protein pattern possessed LPS with a doublet pattern (O1A1) or the lowermost band of the O1A1 doublet (O1A2). O1:K1 strains of the OMP9 pattern possessed LPS referred to as O1A, which corresponded to the uppermost band of the O1A1 doublet pattern. A few O1:K? strains possessed LPS of different migration patterns (O1B and O1C). O1A and O1A1 LPS were indistinguishable by chemical techniques, and both reacted with each of 10 different monoclonal antibodies tested. However, O1A1 had an additional epitope within the additional band in each doublet, as demonstrated by adsorption experiments with hyperimmune rabbit sera followed by Western blotting. Furthermore, purified polysaccharide from O1A bacteria was incapable of inhibition in enzyme-linked immunosorbent assays performed with O1A1 LPS as antigen and adsorbed, specific anti-O1A1 antibodies, whereas O1A1 polysaccharide inhibited this reaction. O1B and O1C LPS differed in all respects tested, including chemical composition, from O1A and O1A1 LPS.
Enhanced primary resistance to Treponema pallidum infection and increased susceptibility to toxoplasmosis in T-cell-depleted guinea pigs
Authors: Pavia C S
Published: January 2020

Strain 2 guinea pigs made T-cell deficient by thymectomy and irradiation and protected with syngeneic bone-marrow cells (TXB guinea pigs) have a surprisingly high level of resistance to cutaneous syphilis and to the dissemination of treponemes to the draining lymph node. Compared with normal euthymic controls infected with Treponema pallidum Nichols, syphilitic TXB guinea pigs developed fewer and less severe skin lesions and their lymph nodes contained lower numbers of treponemes. Associated with this evidence for enhanced innate resistance was the ability of the TXB host to produce, during each test interval of a primary infection, more antitreponemal antibodies than that of their euthymic counterparts. Similar levels of partial protection against cutaneous and disseminated syphilitic infection and elevated antibody levels occurred in challenged normal guinea pigs passively immunized with lymphocytes from T. pallidum-infected TXB donors. In contrast, the capacity of the TXB host to be protected against a lethal infection with the unrelated intracellular protozoan parasite Toxoplasma gondii was greatly impaired unless it received an intravenous infusion of normal syngeneic thymocytes. These seemingly paradoxical results are explained primarily in terms of a residual T-helper-cell population in the TXB guinea pig which is large and competent enough to generate antisyphilis, but not anti-Toxoplasma, immunity.
Preparation of a sialic acid-binding protein from Streptococcus mitis KS32AR
Authors: Murray P A , Levine M J , Reddy M S , Tabak L A , Bergey E J
Published: January 2020

A recent report has identified a lectin on the surfaces of several strains of Streptococcus mitis and Streptococcus sanguis with specificity for an N-acetylneuraminic acid alpha 2,3-galactose-beta 1,3-N-acetylgalactosamine sequence (P.A. Murray, M.J. Levine, L.A. Tabak, and M.S. Reddy, Biochem. Biophys. Res. Commun. 106:390-396, 1982). In the present study, purification and characterization of this sialic acid-binding protein (SABP) was begun. A clinical isolate of S. mitis was grown to mid stationary phase in synthetic FMC medium and then extracted with lithium 3,5-diiodosalicylate. Lyophilized extract was subjected to gel filtration on a Sephadex G-200 column, giving four protein peaks (A to D). Peak B, shown by hemagglutination assay to contain SABP, was next subjected to affinity chromatography on a Sepharose-4B matrix coupled to fetuin glycopeptides. After an extensive washing, peak B materials bound to the affinity matrix were eluted with buffered N-acetylneuraminic acid. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis with 2-mercaptoethanol on 7.5% gels of affinity-purified materials revealed components of 96, 70, and 65 kilodaltons (kDa). Without reducing agent, only the 65-kDa band and materials which did not penetrate the gel were visualized, suggesting that the 96- and 70-kDa components were disulfide linked. The chemical cross-linking agent, disuccinimidyl suberate, was used to demonstrate specific interactions between the SABP preparation and [14C]fetuin glycopeptides. After cross-linking, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography revealed the 96- and 70-kDa components, indicating that the SABP is at least bivalent. These findings support our previous suggestion that human salivary glycoproteins facilitate clearance of selected oral streptococci via specific interactions between sialic acid-containing oligosaccharides and a carbohydrate-binding protein on the bacterial cell surface.
Colonization and infection of athymic and euthymic germfree mice by Campylobacter jejuni and Campylobacter fetus subsp. fetus
Authors: Yrios J W , Balish E
Published: January 2020

Human clinical strains of Campylobacter jejuni and Campylobacter fetus subsp. fetus colonized the gastrointestinal tracts of both athymic (nu/nu) and euthymic (+/nu) germfree mice (BALB/c). Viable Campylobacter spp. (10(9) to 10(10) CFU/g [dry weight] of cecum and colon contents) were isolated on day 3 after oral challenge, and similar large numbers of viable cells were evident at several intervals during a 10-month experiment. The stomachs and upper small intestines of nu/nu and +/nu mice that were monoassociated for 224 days with C. jejuni 45100 contained 3 to 4 logs fewer viable bacteria than did their ceca or colons. Athymic mice that were monoassociated for 224 days with C. fetus subsp. fetus had 2 to 3 logs more viable Campylobacter spp. in their upper gastrointestinal tracts than did their +/nu littermates. Large viable populations (approximately 10(9)/g of contents) of C. fetus subsp. fetus were in the ceca and colons of both nu/nu and +/nu mice. All C. jejuni strains used in this study chronically infected the mesenteric lymph nodes of both nu/nu and +/nu mice. C. jejuni strains 24 and INN 73-83, which were cytotoxic for Chinese hamster ovary cells in vitro, were also more frequently isolated from the livers, spleens, and kidneys of nu/nu mice than was the weak cytotoxin-producing strain 45100. Additionally, heat-labile-enterotoxin-producing C. jejuni INN 73-83 was recovered more frequently from the internal organs of monoassociated +/nu mice than were any other Campylobacter spp. tested. Natural gastrointestinal colonization of neonatal nu/nu and +/nu mice (born to Campylobacter-colonized mothers) with Campylobacter spp. appeared to be delayed until approximately 1 to 2 weeks after birth. Conventionalization of C. jejuni 45100-monoassociated BALB/c mice with a complex mouse fecal microflora eliminated viable C. jejuni from the mesenteric lymph nodes by day 14 and from the cecum by day 78. These findings show that the gnotobiotic BALB/c mouse is a new model for studying acute and chronic host-Campylobacter sp. interactions.
Degradation of protease inhibitors, immunoglobulins, and other serum proteins by Serratia protease and its toxicity to fibroblast in culture
Authors: Molla A , Matsumoto K , Oyamada I , Katsuki T , Maeda H
Published: January 2020

We investigated the effect of the extracellular protease of Serratia marcescens on human serum constituents such as immunoglobulins, fibronectin, alpha 1-protease inhibitor, alpha 2-macroglobulin, lysozyme, and transferrin. At a very low concentration of Serratia 56-kilodalton protease (56K protease), purified human plasma fibronectin was degraded rapidly into three structural domains or small fragments. Immunoglobulin G3 (IgG3) and IgA1 were also degraded within 30 min with 1 microgram of this protease per ml, more rapidly than their other subclass of IgG or IgA. alpha 1-Protease inhibitor, which did not inhibit the 56K protease, was degraded similarly by the protease. These events were demonstrated by fluorescence polarization and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The protease was considerably inhibited by human alpha 2-macroglobulin and chicken ovomacroglobulin. However, when there was a 2 M excess of ovomacroglobulin or a 4 M excess of alpha 2-macroglobulin over the 56K protease, about 25 or 40% proteolytic activity remained, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the protease degraded the alpha 2-macroglobulin extensively during prolonged incubation, which paralleled with regeneration of the protease activity. The protease also cleaved human lysozyme, although moderately. Human serum transferrin was degraded slightly, and human serum albumin was almost resistant to the 56K protease. The enzyme seemed to have no effect on reconstituted collagen, but it degraded rat tropocollagen and yielded fragments of beta and gamma chains by cleaving the intramolecular cross-links. Most of the above proteolysis by the 56K protease appears to result in a limited type of substrate specificity. Thus, the present study demonstrates that the protease is capable of degrading defense-oriented humoral proteins and tissue constituents. Furthermore, it is toxic to fibroblasts. These findings also clarified the possible role of Serratia protease as a virulence factor in the pathogenesis of serratial infections. We recently demonstrated this notion in vivo with rabbit cornea (R. Kamata et al., Ophthalmology 92:1452-1459, 1985).
Enhancement of adherence and growth of Chlamydia trachomatis by estrogen treatment of HeLa cells
Authors: Bose S K , Goswami P C
Published: January 2020

Treatment of HeLa 229 cultures with 17-beta-estradiol or with diethylstilbestrol, a synthetic estrogen analog, prior to infection with Chlamydia trachomatis UW31 (serovar K) or LGV440 (serovar L1) led to a 50 to 60% enhancement of chlamydial inclusion formation. After infection, the presence of estrogen was required for the enhancement. The optimal concentration of estrogen required was 10(-10) M. At least 18 h of preinfection treatment plus 12 h of postinfection treatment was necessary. The adherence of purified radioactive elementary bodies of C. trachomatis to estrogen-treated HeLa cells was stimulated in an estrogen dose- and exposure-dependent manner. The requirements for both pre- and postinfection exposure to the hormone suggest that alterations in the cell membrane as well as in the metabolic capacity of the host cells is required for intracellular chlamydial development. Cycloheximide did not prevent estrogen enhancement of chlamydial adherence or subsequent intracellular development of inclusions.
Functionally active monoclonal antibody that recognizes an epitope on the O side chain of Pseudomonas aeruginosa immunotype-1 lipopolysaccharide
Authors: Stoll B J , Pollack M , Young L S , Koles N , Gascon R , Pier G B
Published: January 2020

A murine monoclonal antibody (MAb) was prepared against Pseudomonas aeruginosa immunotype-1 (It-1) lipopolysaccharide (LPS). The MAb bound It-1 LPS in the enzyme-linked immunosorbent assay and in the immunodiffusion and immunoblotting assays, agglutinated and opsonized It-1 bacteria, and protected against challenge with live It-1 organisms in a murine burn infection model. All of these activities were immunotype specific. Correlation of the opsonic and protective properties of the MAb with its recognition site on the LPS O side chain confirmed that such immunotype-specific determinants are important targets for protective antibodies in Pseudomonas disease. The functional equivalence of this MAb and polyclonal antibodies from hyperimmune plasma underscores the therapeutic potential of single MAbs which recognize critical determinants in the LPS O side chain.
Development of an aerosol model of murine respiratory mycoplasmosis in mice
Authors: Davis J K , Thorp R B , Parker R F , White H , Dziedzic D , D'Arcy J , Cassell G H
Published: January 2020

Animal models of murine respiratory mycoplasmosis due to Mycoplasma pulmonis provide excellent opportunities to study respiratory disease due to an infectious agent. The purpose of the present study was to develop and characterize an aerosol model for the production of murine respiratory mycoplasmosis in mice. The exposure of mice for 30 min to aerosols generated with a DeVilbiss 45 nebulizer in a nose-only inhalation chamber consistently reproduced typical lesions. The chamber was operated with a nebulizer air flow of 5.3 liters/min at 5.0 lb/in and a diluting air flow of 20 liters/min, with the nebulizer containing 5 ml of a suspension of viable M. pulmonis organisms (a concentration between 6 X 10(5) to 6 X 10(10) CFU/ml). Infective aerosol particles of less than a 4.0-micron median aerodynamic diameter with a geometric standard deviation of approximately 2.0 reached the lungs and were evenly distributed among the different lung lobes. A minimum 1.5-log loss of viability in the M. pulmonis suspension was demonstrated. With the exception of the 50% lethal dose, all of the parameters previously established by intranasal inoculation could be examined with the aerosol model. The major advantages of the aerosol model were excellent reproducibility of exposure (both between different experiments and between animals in a given experiment), the avoidance of anesthetization, and the ability to immediately deposit the majority of the organisms in the lung. The only disadvantage was the requirement for large volumes of mycoplasmal cultures.
Lyophilized airborne Clostridium botulinum spores as inocula that intestinally colonize antimicrobially pretreated adult mice
Authors: Sugiyama H , Prather J L , Woller M J
Published: January 2020

Adult mice, made susceptible to Clostridium botulinum by feedings of metronidazole, were immobilized with an anesthetic and held for 30 min in isolators in which a fine powder of lyophilized pathogen spores was made airborne. Exposed mice were surface decontaminated before being kept for 2 days in holding isolators. Mice were intestinally colonized by the pathogen. Colonization rates were related to spore numbers (10(4) to 10(7) type A or B) seeded into isolators.
Antigenic and physical diversity of Neisseria gonorrhoeae lipooligosaccharides
Authors: Mandrell R , Schneider H , Apicella M , Zollinger W , Rice P A , Griffiss J M
Published: January 2020

We used mouse monoclonal antibodies (MAbs) to characterize Neisseria gonorrhoeae lipooligosaccharide (LOS). LOSs that bound two or more MAbs in a solid-phase radioimmunoassay usually bound them to different LOS components, as separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE); strains with multiple LOS components on SDS-PAGE usually bound more than one MAb. However, the LOS of some strains bound the same MAb to two LOS components with different relative molecular weights, and some individual LOS components bound more than one MAb. LOSs from different strains bound different amounts of the same MAb at saturation, reflecting differences in the quantitative expression of individual LOS components. Not all components recognized by MAbs were stained by silver after periodate oxidation. Treatment with NaOH variously affected epitopes defined by different MAbs. MAb 3F11 completely inhibited and MAb 2-1-L8 partially inhibited the binding of 125I-labeled 06B4 MAb to WR220 LOS and WR220 outer membranes in competitive binding studies. Other MAbs did not compete with the binding of 125I-labeled 06B4 to either antigen. We conclude that a strain of N. gonorrhoeae elaborates multiple LOSs that can be separated by SDS-PAGE and that are antigenically distinct. Epitope expression within these glycolipids is complex.
Collagen binding to Staphylococcus aureus
Authors: Holderbaum D , Hall G S , Ehrhart L A
Published: January 2020

Staphylococcus aureus can bind soluble collagen in a specific, saturable manner. We have previously shown that some variability exists in the degree of collagen binding between different strains of heat-killed, formaldehyde-fixed S. aureus which are commercially available as immunologic reagents. The present study demonstrates that live S. aureus of the Cowan 1 strain binds amounts of collagen per organism equivalent to those demonstrated previously in heat-killed, formaldehyde-fixed bacteria but has an affinity over 100 times greater, with Kd values of 9.7 X 10(-11) M and 4.3 X 10(-8) M for live and heat-killed organisms, respectively. Studies were also carried out with S. aureus killed by ionizing radiation, since this method of killing the organism seemed less likely to alter the binding moieties on the surface than did heat killing. Bacteria killed by exposure to gamma radiation bound collagen in a manner essentially indistinguishable from that of live organisms. Binding of collagen to irradiated cells of the Cowan 1 strain was rapid, with equilibrium reached by 30 min at 22 degrees C, and was fully reversible. The binding was not inhibited by fibronectin, fibrinogen, C1q, or immunoglobulin G, suggesting a binding site for collagen distinct from those for these proteins. Collagen binding was virtually eliminated in trypsin-treated organisms, indicating that the binding site has a protein component. Of four strains examined, Cowan 1 and S. aureus ATCC 25923 showed saturable, specific binding, while strains Woods and S4 showed a complete lack of binding. These results suggest that some strains of S. aureus contain high-affinity binding sites for collagen. While the number of binding sites per bacterium varied sixfold in the two collagen-binding strains, the apparent affinity was similar. The ability of S. aureus to bind collagen with high affinity may provide a mechanism for bacterial adhesion to host tissue and thereby play a role in the invasive characteristics of this organism.
Disruption of respiratory cilia by proteases including those of Pseudomonas aeruginosa
Authors: Hingley S T , Hastie A T , Kueppers F , Higgins M L
Published: January 2020

Pseudomonad proteases disrupted the function and structure of demembranated cilia (axonemes) extracted from porcine tracheae. Proteolytic degradation by the two pseudomonad proteases elastase and alkaline protease and by trypsin and subtilisin impaired motility of ATP-activated axonemes. In addition, electron microscopic observation of negatively stained axonemes indicated that exposure to proteases caused dissociation into individual doublet or singlet microtubules. Inhibition of motility and axonemal fraying occurred when axonemes were treated with less than 5 U of proteolytic activity of any of the four proteases tested. When the effects of 2 U of each protease were compared, trypsin and subtilisin were able to produce immotility in less time than pseudomonad elastase and alkaline protease, while alkaline protease and subtilisin caused the most axonemal fraying in 10 min. Proteolytic digestion of axonemal proteins was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. All four proteases cleaved dynein proteins (proteins necessary for motility), though treatment with trypsin resulted in the most extensive solubilization of axonemal proteins. Trypsin and subtilisin both produced more changes in the protein profiles of treated axonemes, using fewer units of proteolytic activity, than the pseudomonad proteases. However, the limited alteration of only a few axonemal proteins by pseudomonad proteases indicates that cleavage need not be extensive to produce dysfunction. Thus, ciliary axonemes are susceptible to proteolytic attack. Degradation of axonemal proteins by pseudomonad proteases, which are released during active infection, may contribute to the impaired ciliary function associated with pseudomonad colonization of the respiratory tract.
Apparent exhaustion of the variable antigen repertoires of Trypanosoma vivax in infected cattle
Authors: Nantulya V M , Musoke A J , Moloo S K
Published: January 2020

Three groups of cattle, each group comprising six animals, were inoculated intravenously with populations of bloodstream forms of Trypanosoma vivax. The first group received T. vivax clone ILDat 1.3 derived from an isolate from Nigeria, while the other two received T. vivax stocks IL 1875 or IL 2133 isolated from Coast Province, Kenya. One animal from the group that was infected with IL 1875 died 8 weeks postinfection. The remaining 17 animals became aparasitemic in 8 to 12 weeks without intervention by drug therapy. The recovered animals developed serodeme-specific immunity against Glossina morsitans subsp. centralis-transmitted challenge. There was complete cross-protection between the two East African T. vivax stocks, although they were isolated from areas 80 to 90 km apart, indicating that they belong to the same serodeme. Antibodies to the homologous metacyclic variable antigen types (VATs) were not detected in sera from recovered animals, suggesting that the immunity displayed by the recovered animals was directed at the bloodstream and not the metacyclic VATs. It is thus suggested that recovery in these animals is due to exhaustion of the repertoire of bloodstream VATs expressed in the animals by the infecting T. vivax clone or stocks.
Correlation of the virulence of Klebsiella pneumoniae K1 and K2 with the presence of a plasmid encoding aerobactin
Authors: Nassif X , Sansonetti P J
Published: January 2020

Nine isolates of Klebsiella pneumoniae belonging to capsular serotypes K1 and K2 were assayed for virulence in mice. Virulent isolates (50% lethal dose of less than 10(3) microorganisms) and avirulent isolates (50% lethal dose of over 10(6) microorganisms) were selected. Supplementation of a defined minimal medium with transferrin markedly reduced the growth of avirulent strains but had no significant effect on the growth of virulent strains. All isolates produced enterochelin, but only production of aerobactin could be correlated with virulence. The genes encoding aerobactin and its receptor protein were located on a 180-kilobase plasmid. They were cloned into the mobilizable vector pSUP202. Homology was demonstrated with the aerobactin operon of the Escherichia coli plasmid pColV-K30. Transfer of the recombinant plasmid pKP4 into an avirulent recipient enhanced virulence by 100-fold. These experiments demonstrated that aerobactin is an essential factor of pathogenicity in K. pneumoniae.
Regulation of production of type 1 pili among urinary tract isolates of Escherichia coli
Authors: Hultgren S J , Schwan W R , Schaeffer A J , Duncan J L
Published: January 2020

The piliation and hemagglutination properties of 54 consecutive Escherichia coli isolates from women with recurrent urinary tract infections were studied. Mannose-sensitive hemagglutination (MSHA) of guinea pig erythrocytes, characteristic of type 1-piliated bacteria, was produced by 75% of the isolates, 32% produced mannose-insensitive hemagglutination, and 14% produced no hemagglutination reaction. The production of type 1 pili was examined in those strains that produced MSHA only. Studies with antiserum prepared against purified pili suggested that at least three subtypes of type 1 hemagglutinins were represented among the isolates. All of the type 1-piliated isolates produced MSHA after serial subculture in static broth. After growth on agar, selected type 1-piliated isolates were subdivided into two groups. Many strains apparently suppressed piliation during growth on agar (regulated variants); all colonies became MSHA negative and were composed of nonpiliated cells as shown by electron microscopy. The loss of the MSHA phenotype often occurred after a single overnight passage on agar, and any remaining hemagglutinin was gradually lost with one to three additional passages. Seven strains, however, retained a significant hemagglutination titer after multiple subcultures on agar, and they produced colonies consisting of a mixed population of piliated and nonpiliated cells. These strains were apparently able to oscillate between states of pilus expression and nonexpression during growth on agar (random phase variants). When nonpiliated cells isolated from the mixed, random variant population were plated on agar, they gave rise to hemagglutination-positive colonies that consisted of both piliated and nonpiliated cells. The distinction between random variants and regulated variants was also observed in shaking broth cultures inoculated with nonpiliated cells. The random variants produced MSHA-positive cultures composed of piliated and nonpiliated cells, whereas the regulated strains remained nonpiliated. The results indicate that type 1 pili are a predominant adhesin of uropathogenic E. coli and that during growth on agar only about one-fourth of the type 1-piliated isolates regulate pilus expression by random phase variation.