bluuebunny/crossref_metadata_2025 huggingface dataset:
Antibodies to cell envelope proteins of Pseudomonas aeruginosa in cystic fibrosis patients
Authors: Fernandes P B , Kim C , Cundy K R , Haung N N
Published: January 2020

Many vaccines containing somatic and secreted antigens of Pseudomonas aeruginosa have been reported. The vaccines containing lipopolysaccharide have been found to provide type-specific protection, but the endotoxin content of these vaccines does not make it feasible to use them in patients who are already debilitated. Outer membrane proteins could be effective as vaccines, as they can be purified free of lipopolysaccharide, and also because they are common to all serotypes of P. aeruginosa. To be effective as a vaccine, such proteins must be immunogenic and accessible from the outside of the intact bacterial cell. In this study, we showed that systemic antibodies were produced frequently to two cell envelope proteins with masses of 58,500 and 37,500 daltons and occasionally to 34,000-dalton protein of P. aeruginosa in cystic fibrosis patients with chronic lung infections. In rabbits immunized with whole, fixed cells of P. aeruginosa, antibodies were also produced against the 58,500-dalton proteins. Thus, the 58,500-dalton cell envelope protein of P. aeruginosa was the only immunogenic protein that was accessible to the immune system when whole, fixed cells were used for immunization. These serum antibodies did not protect the cystic fibrosis patients against further lung infection with P. aeruginosa.
Phagocytosis of Giardia muris by macrophages in Peyer's patch epithelium in mice
Authors: Owen R L , Allen C L , Stevens D P
Published: January 2020

No mechanism for the initiation of immunological clearance of Giardia from the mammalian intestinal tract has been identified. In normal and nude mice experimentally infected with G. muris, we examined antigen-sampling epithelium over Peyer's patch follicles by electron microscopy for evidence of interaction between G. muris and lymphoid cells. Invading G. muris were found in the epithelium near dying or desquamating columnar cells. Macrophages beneath the basal lamina extended pseudopods into the epithelium, trapping invading G. muris and enclosing them in phagolysosomes. In normal mice, which clear G. muris in 4 to 6 weeks, macrophages containing digested G. muris were surrounded by rosettes of lymphoblasts in the epithelium. In nude mice deficient in lymphocytes, there was apparent hyperplasia of macrophages, which filled the follicle domes, resulting in more frequent entrapment of G. muris but no contact between macrophages and lymphoblasts in the epithelium. In nude mice, which require 6 months to control G. muris infection, lymphoblast contact with macrophages containing distinctive microtubular remnants of G. muris was only identified in the follicle dome. This close physical association of lymphoblasts and macrophages containing G. muris remnants suggests that this macrophage activity represents intraepithelial antigen processing as well as a defense against the effects of the uncontrolled entrance of microorganisms and other antigenic particles into Peyer's patch lymphoid follicles.
Toxicities of
Clostridium botulinum
Type C Toxins of Different Molecular Sizes in Geese
Authors: Ohishi Iwao , Hayashi Ken-Ichi , Sakaguchi Genji , Tokuchi Mikio
Published: January 2020

Clostridium botulinum
type C toxin of 16S was more potent as an oral toxin to geese than that of 12S.
Binding of human immunoglobulin G to protein A in encapsulated Staphylococcus aureus
Authors: King B F , Wilkinson B J
Published: January 2020

Recent studies of the mechanism of resistance to phagocytosis in encapsulated Staphylococcus aureus have suggested that the capsule is readily penetrated by high-molecular-weight proteins such as antibodies and complement components. S. aureus strains contain a cell wall protein, protein A, that reacts with the Fc portion of immunoglobulins. The binding of immunoglobulin G (IgG) to encapsulated and unencapsulated S. aureus strains has been studied to assess the penetrability of the S. aureus capsule by IgG. Encapsulated S. aureus strains M and Smith diffuse bound large amounts of human IgG which were comparable to amounts bound by the unencapsulated strains Cowan I, M variant, and Smith compact. Trypsin treatment of bacteria reduced their ability to bind IgG. Bound IgG was not removed by extensive washing of bacteria with buffer. A non-protein A-containing, coagulase-negative, encapsulated staphylococcal strain did not bind IgG. These observations suggest that IgG is binding to cell wall protein A in encapsulated S. aureus. No differences in the rates of IgG binding by encapsulated and unencapsulated S. aureus strains were observed. It is concluded that the S. aureus capsule is freely permeable to IgG. This is of importance in considerations of the mechanisms of resistance to phagocytosis and antigen masking in encapsulated microorganisms.
Toxicity of Haemophilus pleuropneumoniae for porcine lung macrophages, peripheral blood monocytes, and testicular cells
Authors: Bendixen P H , Shewen P E , Rosendal S , Wilkie B N
Published: January 2020

Strain CM5 of Haemophilus pleuropneumoniae was toxic for porcine lung macrophages in minimum doses of 10(6) colony-forming units per ml and for peripheral blood monocytes at 10(7) colony-forming units per ml. The organism was not toxic for porcine testicular cells in a concentration of 10(8) colony-forming units per ml. Filtered sterile culture supernatant diluted 10(0), 10(-1), and 10(-2) was toxic for porcine pulmonary lavage cells. In dilutions of 10(0) and 10(-1), culture supernatant was toxic for peripheral blood monocytes, and at 10(0) it was toxic for testicular cells. Toxicity associated with bacterial cells was sensitive to heating (60 degrees, 60 min), whereas that of the culture supernatant was heat stable (100 degrees C, 15 min). Swine convalescent serum neutralized supernatant toxicity in dilutions of greater than or equal to 1:320. The findings promote the understanding of the pathogenesis of pleuropneumonia in swine caused by H. pleuropneumoniae.
Subunit number and arrangement in Escherichia coli heat-labile enterotoxin
Authors: Gill D M , Clements J D , Robertson D C , Finkelstein R A
Published: January 2020

The Escherichia coli heat-labile enterotoxin (LT) was found to have the same subunit structure as cholera toxin, namely, one A subunit and five B subunits. Reaction with a bisimidate generated all the possible cross-linked derivatives of A5B: B,2B ... 5B and A, AB ... A5B. The isolated B component, coligenoid, contained five B subunits and showed some tendency of polymerize: with a bisimidate it became covalently connected into the set B ... 5B with lesser amounts of 6B ... 10B, etc. The subunit formulas of two independently prepared samples of LT were both proved to be A5B by cross-linking, but their B pentamers migrated at different rates on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, indicating that they have different conformations. The faster (R) form could be converted to a diffuse slower (C) form by incubating it at 50 degrees C or at 37 degrees C with 0.2 M galactose, which is the terminal sugar of ganglioside GM1, the natural receptor for LT. Cholera toxin resembled the R form more than the C form of LT.
Fibronectin binding to protein A-containing staphylococci
Authors: Doran J E , Raynor R H
Published: January 2020

Fibronectin (Fn) was found to bind to protein A-containing isolates of Staphylococcus aureus, but not to mutant strains devoid of this protein nor to clinical isolates of S. epidermidis. Fn was purified from human plasma by affinity chromatography on gelatin-Sepharose. After elution with 4 M urea, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified material detected no immunoglobulin contamination. This purified Fn was radiolabeled with 125I and used in binding assays. Quantitatively, Fn binding was directly correlated with the cellular protein A content of the various strains tested. Mannitol salt broth preculture or organisms resulted in a reduction of their cellular protein A and a decrease in Fn binding by these cells. However, soluble protein A maximally inhibited the binding of radiolabeled Fn to protein A-positive strains of staphylococci by only 50%, indicating the possibility of multiple Fn binding sites. Fn's binding to protein A-containing S. aureus strains may play a role in the pathogenicity of these organisms by promoting their attachment to and subsequent invasion of host tissues.
In vitro cellular immune response to homologous and heterologous antigens in rabbits sensitized by five species of Mycobacterium and Nocardia asteroides
Authors: Kubín M , Pekárek J , Svejcar J , Procházka B
Published: January 2020

Rabbits were sensitized with killed Mycobacterium tuberculosis, M. kansasii, M. avium, M. scrofulaceum, M. fortuitum, and Nocardia asteroides, and their response to homologous and heterologous antigens was assessed in vitro by direct and indirect macrophage migration inhibition tests. The antigens were obtained by disintegration of bacterial mass and by purification of the supernatants by ultracentrifugation. In the direct test, hypersensitivity to homologous antigen was most marked with M. tuberculosis, M. kansasii, and M. avium (migration indices [MI] = 0.42 to 0.50), but was significantly weaker with organisms possessing a lower degree of pathogenic activity (MI for M. fortuitum and N. asteroides = 0.70 and 0.72, respectively). Reactivity to heterologous antigens was also highest in animals sensitized with strongly pathogenic species, approximating normal values in rabbits sensitized with weak pathogens. In the indirect test, the strongest responses were obtained again to homologous antigens (MI = 0.42 to 0.67), and they differed more markedly from reactions to heterologous antigens than in the direct test. The weakest activity of heterologous antigens was again found with M. fortuitum and N. asteroides, where MIs were 0.82 to 0.93.
Recovery of mice from herpes simplex virus type 2 hepatitis: adoptive transfer of recovery with immune spleen cells
Authors: Mogensen S C , Andersen H K
Published: January 2020

Young BALB/c mice inoculated intraperitoneally with herpes simplex virus type 2 develop focal necrotizing hepatitis. After infection, the livers of these mice show increasing virus titers, which reach a maximum on day 3 after infection; this is followed by a dramatic decrease in the amount of virus recovered on days 4 and 5. This decrease in virus content is accompanied by a progressive infiltration of the lesions with mononuclear leukocytes and an apparent resolution of the lesions. Adoptive transfer of immune spleen cells from mice infected 6 days earlier accelerated this process. When 50 x 10(6) to 100 x 10(6) immune spleen cells were transferred 24 h after infection, the inflammatory response and the clearance of virus from the livers were advanced by almost 2 days. As few as 12 x 10(6) immune spleen cells accelerated the healing process, whereas fewer immune cells, disrupted immune cells, or normal spleen cells did not have an effect. The protection conferred by herpes simplex virus type 2-sensitized immune spleen cells was specific since mouse cytomegalovirus- or vaccinia virus-sensitized immune spleen cells had no effect on the course of infection with herpes simplex virus type 2, whereas some cross-reactivity was observed between herpes simplex virus types 1 and 2. This model seems to be suitable for examining the immunological mechanisms that are active during recovery from visceral herpes simplex virus infections.
Radioimmunoassay for human beta interferon
Authors: Inoue M , Tan Y H
Published: January 2020

Pure human beta interferon (Hu IFN-beta) (formerly human fibroblast interferon) was used to raise antibodies in rabbits. Anti-IFN-beta immunoglobulins from rabbit serum were purified by IFN-beta affinity chromatography and were used in the development of a sandwich-type radioimmunoassay for Hu IFN-beta. This radioimmunoassay consisted of (i) incubating Hu IFN-beta with anti-IFN-beta immunoglobulins immobilized on Sepharose 4B beads and (ii) incubating the resulting solid-phase IFN antibody complex with 125I-labeled anti-Hu IFN-beta immunoglobulins. The amount of 125I-labeled anti-Hu IFN-beta bound to the complex was linearly proportional to the amount of antiviral activity present in the complex from 0 to 500 U of standard IFN-beta per ml; 1 U of antiviral activity was equivalent to about 40 cpm of 125I-labeled anti-IFN immunoglobulins in the radioimmunoassay. The results of the radioimmunoassay were not influenced by the protein content of the IFN sample assayed, and the degree of intraassay variability was low; coefficients of variability were 9.4% at 5 U/ml and 1.1% at 500 U/ml. Thus, compared with the traditional antiviral assay for IFN, this radioimmunoassay was equally sensitive and more precise. In addition, it was highly specific for Hu IFN-beta and took only 5 h to complete.
Humoral immunity and reduced periodontal bone loss in Eikenella corrodens-monoassociated rats
Authors: Behling U H , Sallay C , Sanavi F , Pham P H , Nowotny A
Published: January 2020

Germfree Sprague-Dawley rats monoassociated with Eikenella corrodens exhibited alveolar bone loss. This progressive bone loss occurred over a period of weeks, during which time the hosts developed an immune response toward the infective microorganism. By means of repeated bacterial vaccination resulting in elevated serum antibody titers, reduced bone loss was observed.
Effects of cell source, mouse strain, and immunosuppressive treatment on production of virulent and attenuated murine cytomegalovirus
Authors: Selgrade M K , Nedrud J G , Collier A M , Gardner D E
Published: January 2020

Murine cytomegalovirus pools from various in vitro and in vivo sources were compared for virulence in suckling mice in an effort to identify the conditions which were necessary for the production of virulent and attenuated viruses. Virus passaged in tracheal ring and salivary gland organ cultures, where virus is produced primarily by epithelial cells, was even more attenuated than virus passaged in mouse embryo fibroblasts. The attenuation observed after passage in all three of these in vitro systems did not appear to be due to defective interfering particles. We also found that virus produced in vivo in salivary glands became attenuated with time after infection. Virus harvested from salivary glands 5 to 6 weeks after infection was highly attenuated compared with both salivary gland-passaged virus harvested 2 to 3 weeks after infection and tissue culture-passaged virus. The attenuation of salivary gland-passaged virus with time was reversed when animals were treated with cyclophosphamide before the virus was harvested. A comparison of virus pools harvested from susceptible and resistant mouse strains indicated that the mouse strain had little effect on the virulence of the virus produced. When the various sources of virus tested in this study were ranked in terms of the virulence of the virus produced, salivary glands in intact mice either 2 to 3 weeks after infection or after cyclophosphamide treatment produced the most virulent virus, followed by mouse embryo fibroblast cultures, tracheal ring and salivary gland organ cultures, and, finally, salivary glands in intact mice 5 to 6 weeks after infection.
Purification, characterization, and partial covalent structure of Escherichia coli adhesive antigen K99
Authors: de Graaf F K , Klemm P , Gaastra W
Published: January 2020

The adhesive antigen K99 of enterotoxigenic Escherichia coli strains of calf origin was isolated and purified. The K99 fimbriae were removed from the cells by heat treatment, concentrated by precipitation with ammonium sulfate, and purified by gel filtration on Sepharose CL-4B and treatment with deoxycholate. The purified K99 antigen was composed of protein subunits with a molecular weight of 18,500 and had an isoelectric point of 9.5. The N-terminal amino acid sequence, as well as the composition of the C-terminal part of the K99 protein subunits, was determined.
Construction and expression of recombinant plasmids encoding type 1 or D-mannose-resistant pili from a urinary tract infection Escherichia coli isolate
Authors: Hull R A , Gill R E , Hsu P , Minshew B H , Falkow S
Published: January 2020

Isolates of Escherichia coli from human urinary tract infections frequently express adherence properties found less often among normal intestinal isolates. These properties include adherence to human uroepithelial cells and primary monkey kidney cells, as well as D-mannose-resistant hemagglutination of human erythrocytes, and they are mediated by a pilus type different from type 1. The genes encoding this pilus type (pyelonephritis-associated pili, pap) and those encoding type 1 pili have been cloned from a urinary tract infection isolate of E. coli and transferred to an E. coli K-12 derivative. The recombinant plasmids were found to express functional pili and to endow the new host with all of the adherence properties of the urinary tract infection isolate. Both pilus types were found to be genetically distinct, and unlike the adherence genes from bovine, porcine, and human diarrheal isolates, both were found to be chromosomally encoded.
Rabbit macrophages secrete two biochemically and immunologically distinct endogenous pyrogens
Authors: Murphy P A , Cebula T A , Levin J , Windle B E
Published: January 2020

Rabbit endogenous pyrogens occurred in two forms. One was an apparently single protein with a pI of 7.3; the other was a family of proteins with pI values of 4.5 to 5.0. We selected two of the latter, with pI values of 4.6 and 4.72, as representative of the group and compared them with the pI 7.3 pyrogen. Antisera raised in three goats completely neutralized the pyrogenic activity of the pI 7.3 pyrogen. Larger doses of these antisera did not block the pyrogenic activity of either of the pI 4.5 to 5.0 pyrogens. The pI 7.3 pyrogen contained a free --SH group which was essential to its biological activity. It was inactivated by 100 mM N-ethylmaleimide or 200 mM iodoacetamide, bound to Thiol-Sepharose columns, and could be eluted from them with mercaptoethanol. Neither of the pI 4.5 to 5.0 pyrogens was inactivated by N-ethylmaleimide or iodoacetamide, and neither bound to Thiol-Sepharose. Both endogenous pyrogens gave negative results in the Limulus lysate test for bacterial endotoxins. These results suggest that the pI 7.3 and pI 4.5 to 5.0 endogenous pyrogens are not closely related to each other and are consistent with the idea that they may not be related at all. Alternative hypotheses are discussed.
Use of monoclonal antibody directed against herpes simplex virus glycoproteins to protect mice against acute virus-induced neurological disease
Authors: Dix R D , Pereira L , Baringer J R
Published: January 2020

Monoclonal antibodies HCl and HD1, directed against herpes simplex virus type 1 (HSV-1) glycoproteins gC and gD, respectively, were evaluated for their ability to passively immunize mice against acute virus-induced neurological disease after footpad inoculation with HSV-1 or herpes simplex virus type 2 (HSV-2). Control virus-infected mice receiving a single intraperitoneal injection of normal serum died within 7 to 10 days after the spread of virus from footpad to spinal cord and brain. However, a single intraperitoneal injection of either HCl or HD1 antibody protected mice from neurological illness and death when administered to HSV-1 (strain HTZ)-infected mice at either 2 h before virus challenge or at 24 h after virus inoculation. To determine the in vivo specificity of the antibodies, passive transfer studies were performed with mice infected with the MP strain of HSV-1, a mutant of HSV-1 (mP) which is defective in the production of glycoprotein gC. Whereas HD1 antibody decreased the incidence of neurological illness in MP- and mP-infected mice, HCl antibody, which protected mP-infected animals, failed to protect mice infected with the MP strain. When HD1 antibody was administered to HSV-2 (strain G)-infected mice at either 2 h before virus challenge or at 6 h (but not 24 h) after virus inoculation, 100% of the infected animals receiving HD1 antibody survived. In contrast, 100% of HSV-2 (strain G)-infected animals passively immunized with HCl antibody developed neurological illness and died. These results provide in vivo evidence that the HSV-induced glycoprotein gC expresses type-specific antigenic determinants, whereas glycoprotein gD expresses type-common determinants.
Experimental Campylobacter diarrhea in chickens
Authors: Ruiz-Palacios G M , Escamilla E , Torres N
Published: January 2020

An animal model for Campylobacter fetus subsp. jejuni enteritis was developed in 3-day-old chickens. Diarrhea was induced in 88% (22 of 25) of chickens inoculated with 9 X 10(7) bacteria given orally. The mean incubation time was 45 h (range, 24 to 72 h). Considerable weight loss was observed in the experimental group compared with the control group. Ninety bacteria was the minimal infective dose capable of inducing diarrhea in 90% of the chickens. Overall mortality was 32% (8 of 25). Light microscopy, immunofluorescence, and electron microscopy of the gastrointestinal tract of serially sacrificed chickens were performed in control and experimental groups. A moderate infiltration of mononuclear cells was observed in ileum and cecum in the experimental group, with no disruption of intestinal mucosa. By immunofluorescence and electron microscopy, campylobacter was located within the epithelial cells and phagocytosed to a greater degree by mononuclear cells of the lamina propria.
Congenital infection of mice with Japanese encephalitis virus
Authors: Mathur A , Arora K L , Chaturvedi U C
Published: January 2020

Transplacental transmission of Japanese encephalitis virus (JEV) when given intraperitoneally was demonstrated in pregnant mice as shown by isolation of the virus from placenta and fetal tissues. Furthermore, JEV could be isolated from the brain, liver, and spleen of newborn mice. The effect of JEV at different periods of gestation in pregnant mice was demonstrated for the first time, and the consequences of maternal infection on fetuses and neonates were studied. JEV infection during the 1st week of gestation caused a significantly higher number of fetal and neonatal deaths (66%) than during the 3rd week of gestation (13.8%). The number of abortions, stillbirths, and neonatal deaths was higher in infected mothers than in controls. No congenital abnormalities were found in any of the newborn mice. Sera obtained from 5-week-old health mice delivered by mothers infected during the 3rd week of gestation contained JEV hemagglutination inhibiting and immunoglobulin M antibodies. The results of these preliminary experiments show the usefulness of mice as a model for further elucidation of JEV infection during pregnancy and its effects on the fetus.
Parainfluenza 3 virus: plaque-type variants lacking neuraminidase activity
Authors: Shibuta H , Kanda T , Hazama A , Adachi A , Matumoto M
Published: January 2020

Virus clones lacking detectable neuraminidase activity (SC-YN and M-YN) as well as those possessing it (LT-910N and LT-YN) were isolated from bovine strains of parainfluenza 3 virus. LT-910N and LT-YN viruses produced large turbid plaques in MDBK cells, and SC-YN virus produced small clear plaques. Incorporation of a bacterial neuraminidase in agar overlay medium made SC-YN virus form large turbid plaques, whereas it made M-YN virus form large clear plaques. However, M-YN virus formed only pinhole plaques or no plaques in the absence of neuraminidase. The exogenous neuraminidase had little effect on the plaque formation of LT-910N and LT-YN viruses. M-YN virus induced extensive syncytial formation, and SC-YN virus produced less extensive syncytial formation. The exogenous neuraminidase enhanced replication of SC-YN and M-YN viruses and reduced syncytial formation by these viruses. The enzyme had little effect on replication and cytopathic effect of LT-910N and LT-YN viruses. The reason for these effects of the exogenous neuraminidase is discussed.
Activation of the Alternative Complement Pathway by Blood Culture Isolates of
Bacteroides fragilis
Authors: Joiner Keith A. , Hawiger Alexandra , Gelfand Jeffrey A.
Published: January 2020

Blood culture isolates of
Bacteroides
caused depletion of alternative complement pathway hemolytic activity, but no fall in C4 titers, when incubated in vitro with normal serum or serum containing 0.002 M MgCl
2
and 0.008 M ethylene glycol-bis(β-aminoethyl ether)-
N,N
-tetraacetic acid.
Production of cholera-like enterotoxin by a Vibrio cholerae non-O1 strain isolated from the environment
Authors: Craig J P , Yamamoto K , Takeda Y , Miwatani T
Published: January 2020

Vibrio cholerae non-O1 strain E8498, isolated in 1978 from fresh water in Louisiana, produced a vascular permeability factor when cultured in shallow resting cultures of Casamino Acids-yeast extract-glucose medium for 24 h at 30 degrees C. Undiluted resting culture filtrates contained heat-labile permeability factor activity which was only partially neutralized by cholera antitoxin and GM1 ganglioside. Supernatants concentrated with PM-10 membranes caused hemorrhage and necrosis in rabbits within 1 h after intracutaneous injection, whereas appropriate dilutions of both filtrates and concentrates demonstrated delayed permeability factor activity, without hemorrhage or necrosis, which was indistinguishable in appearance from that caused by purified cholera enterotoxin produced by V. cholerae O1 Inaba strain 569B. Crude E8498 filtrates contained the biological equivalent of about 5 ng/ml of purified enterotoxin. Permeability factor activity in the fraction obtained by 20 to 50% saturation of filtrate concentrate with ammonium sulfate could be completely neutralized by reference standard cholera antitoxin prepared against purified 569 B enterotoxin. Hemorrhagic activity was unaffected by cholera antitoxin. A 5,000-fold concentrate of the culture supernatant yielded a line of identity with purified cholera enterotoxin in an agar gel double-diffusion test against cholera antitoxin purified by affinity column chromatography with BrCN-activated Sepharose 4B-linked purified cholera enterotoxin as the adsorbent. These findings indicate that V. cholerae non-O1 E8498 produces a permeability factor which is immunologically and biologically indistinguishable from that produced by a strain of V. cholerae O1 classical biotype.
Role of lymphocytes in macrophage-induced killing of Coccidioides immitis in vitro
Authors: Beaman L , Benjamini E , Pappagianis D
Published: January 2020

Peritoneal macrophages from normal mice phagocytized arthroconidia and endospores of Coccidioides immitis without affecting the viability of the spores within 4 h after infection. In contrast, macrophages, when infected in the presence of lymphocytes from immune mice, significantly reduced the viability of phagocytized endospores and arthroconidia. The inability of macrophages from normal mice to kill C. immitis may in part be explained by the observation that C. immitis appeared to inhibit fusion of the phagosomes containing fungal spores with the lysosomes within the macrophages. However, fusion of phagosomes containing spores and lysosomes was observed in macrophages infected in the presence of lymphocytes from immune mice.
Macrophage function during Trypanosoma musculi infection in mice
Authors: Vincendeau P , Caristan A , Pautrizel R
Published: January 2020

Quantitative and functional changes in peritoneal macrophages from mice infected with Trypanosoma musculi were investigated. Increase in the number and size and in the protein content of peritoneal macrophages and the presence of parasites in the peritoneal activity were observed during the course of parasitemia. Induced releases of H2O2 by macrophages were increased during parasitemia, but H2O2 release returned to normal after infection. A correlation was also observed between parasitemia and 99mTc colloidal uptake by peritoneal macrophages. These changes in macrophage function may indicate an activation of these cells.
Homogeneity and Heterogeneity of Toxins Produced by
Clostridium botulinum
Type C and D Strains
Authors: Oguma K. , Syuto B. , Agui T. , Iida H. , Kubo S.
Published: January 2020

Five
Clostridium botulinum
strains were used in the present work, two of type C, C-Stockholm (C-ST) and C-CB19, and three of type D, D-South African (D-SA), D-1873, and D-CB16. The toxins, except for those of C-CB19 and D-CB16, were purified, and antisera were prepared in rabbits. To clarify the antigenicity of the toxins, neutralization and agar gel double-diffusion tests were performed. Anti-C-ST toxin serum neutralized two kinds of type C
1
toxin to a similar extent. Antisera against D-SA and D-1873 toxins, however, showed different neutralizing activity toward three type D toxins. Precipitin lines formed between D-SA and D-1873 toxins, and their antisera spurred to each other. From anti-D-SA toxin serum, two neutralizing fractions, one which neutralized D-SA, D-1873, and D-CB16 and one which neutralized only D-SA, were obtained. These results indicate that the antigenicities of D-SA and D-1873 toxins are not identical. Anti-C-ST toxin serum produced cross-neutralization on type D toxins, although the neutralization titer differed depending on the kind of toxin used. A precipitin line was formed with D-SA toxin, but not with D-1873; the developed line spurred to the C-ST toxin precipitin line. Two anti-D toxin sera also caused cross-neutralization on type C
1
toxins. However, the neutralizing activity of each serum to the same type C
1
toxin was different, and only anti-D-SA toxin serum developed a precipitin line with C-ST toxin which spurred to the D-SA toxin precipitin line. From anti-D-SA toxin serum, two different fractions capable of neutralizing C
1
and D toxins were obtained; one neutralized C-ST, C-CB19, and D-SA toxins, but not D-1873 and D-CB16, and the other neutralized all five toxins. There may be at least two common parts among C-ST, C-CB16, and D-SA toxin molecules.
Isolation of a protein-containing cell surface component from Streptococcus sanguis which affects its adherence to saliva-coated hydroxyapatite
Authors: Liljemark W F , Bloomquist C G
Published: January 2020

The isolation and partial characterization of a protein-containing cell surface component from Streptococcus sanguis which blocks the adherence of this microbe to saliva-coated hydroxyapatite are described. Several methods of extraction were attempted. Sonication of whole cells and cell walls proved to be the most successful and yielded biologically active adherence-blocking components. The adherence-blocking ability of these components was effective in intraspecies blocking experiments. The extract obtained from cell walls of S. sanguis was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and shown to contain one major and two to three minor bands when stained with Coomassie blue. The molecular weight of the major band was estimated to be 70,000 to 90,000. Gel filtration of the sonified cell wall extract on 10% agarose yielded two active adherence-blocking peaks, the void volume and a second peak.
Protein A effect on alternative pathway complement activation and opsonization of Staphylococcus aureus
Authors: Spika J S , Verbrugh H A , Verhoef J
Published: January 2020

Twelve Staphylococcus aureus strains with known amounts of protein A were compared with regard to alternative pathway complement activation and opsonization in human serum. "Protein A-poor" strains (less than or equal to 0.16 ng/10(6) bacteria) were, on the average, 3. 4-fold more efficient in alternative pathway complement activation than "protein A-rich" strains (greater than or equal to 0.625 ng/10(6) bacteria) (P less than 0.001). Protein A-poor strains were significantly better phagocytized by human polymorphonuclear leukocytes after opsonization in magnesium-ethylene glycol-bis (beta-amino-ethyl ether)-N, N-tetraacetic acid-chelated serum than were the protein A-rich strains (P less than 0.001). No significant differences between protein A-poor and -rich strains were found in complement activation and opsonization in normal serum. Cell wall-bound protein A appeared to hinder alternative pathway complement activation by S. aureus, which resulted in decreased opsonization of these bacteria in the absence of an intact classical pathway. These studies suggest that protein A may cover alternative pathway complement-activating sites within the peptidoglycan matrix of the staphylococcal cell wall.
High-molecular-weight polysaccharide antigen from Pseudomonas aeruginosa immunotype 2
Authors: Pier G B , Sidberry H F , Sadoff J C
Published: January 2020

Previously, we isolated a high-molecular-weight immunogenic polysaccharide (designated PS) from Pseudomonas aeruginosa immunotype 1 (IT-1). The method which we used was modified to permit the isolation of a similar PS from P. aeruginosa IT-2. This antigen was composed primarily of carbohydrate, had a complex monosaccharide composition, including sugars not found in the lipopolysaccharide, and was nonpyrogenic in rabbits and nontoxic in mice at high doses. This material protected mice from challenges with live homologous cells. P. aeruginosa IT-2 PS gave a line of identity with the O side chain of the lipopolysaccharide, but different from this polysaccharide in molecular weight, chemical composition, and ability to immunize mice actively. Lipopolysaccharide from P. aeruginosa IT-2 contained an immunological determinant not found on P. aeruginosa IT-2 PS, which was detected due to its stability during treatment with dilute alkali. Thus, we recovered a high-molecular-weight PS antigen from P. aeruginosa IT-2, which was serologically identical to the lipopolysaccharide O side chain but was chemically and physically distinct. Also, like P. aeruginosa IT-1 strains, P. aeruginosa IT-2 contains an alkali-stable immunodeterminant on the lipopolysaccharide that may represent a core-like antigen.
Spontaneous and induced herpesvirus genome expression in Marek's disease tumor cell lines
Authors: Calnek B W , Shek W R , Schat K A
Published: January 2020

We incubated 31 newly established Marek's disease tumor cell lines at 41 degrees C for 48 h after subculturing and then examined them to determine the spontaneous rates of expression of viral internal antigen(s), viral membrane antigen(s), and virus isolation. All but two of the lines were isolated from tumors induced by clone-purified Marek's disease virus strain JM-10, GA-5, RB-1B, and BC-1A in nine different genetic strains of chickens with defined histocompatibility antigens. The line-to-line variations in the rates of spontaneous expression for the antigens or virus rescue were great, but the levels of expression were very low in most cases. The median rates of expression for viral internal antigen, viral membrane antigen, and virus isolation were 32, 8, and 2 positive cells per 10(5) cells, respectively (ranges, 0 to 20,280, 0 to 22,990, and 0 to 220 positive cells per 10(5) cells, respectively). The ratio of viral internal antigen expression to virus isolation was extremely variable and often high, whereas the ratio of viral internal antigen to viral membrane antigen expression was more consistent and generally low. The virus strain which induced the cell line influenced the level of virus genome expression, but the cell genotype did not. Cell lines transformed by JM-10 virus, which exhibited low oncogenicity, had significantly (p less than 0.01) higher rates of expression than cell lines transformed by CA-5 and RB-1B viruses, which exhibited high oncogenicity. Treatment with iododeoxyuridine or incubation at 37 degrees C induced increased rates of expression in most lines but not in all lines. The degree of enhanced expression was inversely proportional to the rate of spontaneous expression.
Bactericidal capacity of newborn phagocytes against group B beta-hemolytic streptococci
Authors: Becker I D , Robinson O M , Bazán T S , López-Osuna M , Kretschmer R R
Published: January 2020

The bactericidal capacity of mononuclear and polymorphonuclear phagocytes obtained from normal newborn infants and from healthy adults was evaluated in vitro, using two group B beta-hemolytic streptococci (GBBHS) serotypes (GBBHS-Ia-SS-615/28 and GBBHS-III-SS-620/50) and uniform opsonic conditions. No intertype differences in bacteriolysis of these two serotypes were observed among leukocytes from newborns or adults. As group, only polymorphonuclear phagocytes from newborns disclosed a significantly lower mean bactericidal capacity than their adult cellular counterpart, and only with respect to GBBHS-III-SS-620/50. On the other hand, 4 or 16 polymorphonuclear samples from newborns tested revealed significantly low bactericidal capacities against both GBBHS serotypes, and an additional sample revealed a bactericidal capacity against GBBHS-III-SS-620/50 alone. Interstrain variations in the intrinsic bactericidal capacity of polymorphonuclear phagocytes from newborns against GBBHS-III may exist, as suggested by a single observation made by using four clinical isolates of GBBHS-III. Such deviant phagocytic capacities of polymorphonuclear phagocytes from newborns may constitute an additional selective risk factor in the genesis of GBBHS sepsis of the newborn.
Localization of chlamydial group Antigen in McCoy cell monolayers infected with Chlamydia trachomatis or Chlamydia psittaci
Authors: Richmond S J , Stirling P
Published: January 2020

Chlamydial inclusions were demonstrated by indirect immunofluorescence (IF) with antiserum to the chlamydial group antigen when McCoy cell monolayers infected with either Chlamydia trachomatis or Chlamydia psittaci were fixed in formaldehyde or paraformaldehyde, provided the monolayer was not allowed to dry. If these monolayers were then air dried and restained by IF with the same antiserum but with a different fluorescence conjugate, group antigen associated with inclusion-containing McCoy cells but independent of the inclusions was revealed. This antigen was not restricted to infected cells but appeared to radiate out from them, suggesting that group antigen was released from infected cells. Similar host cell-associated antigen could be shown by IF of glutaraldehyde-fixed, air-dried monolayers, but inclusions could not be stained by IF before these preparations were dried, presumably because antibody could not penetrate glutaraldehyde-fixed cells. Electron microscopic immunoperoxidase studies of paraformaldehyde-fixed, wet monolayers located group antigen within inclusions on the outer membrane of chlamydial organisms and on single-membrane vesicles. However, when dried monolayers were labeled with the same immunoperoxidase technique, no intracellular labeling occurred, but dense staining was seen at the surface of infected cells and on adjacent membranous material. These observations are compatible with the postulate that replicating chlamydiae produce outer membrane blebs containing group antigen, which are excreted by the host cells during the chlamydial developmental cycle.
Effects of lectins on peripheral infections by herpes simplex virus of rat sensory neurons in culture
Authors: Ziegler R J , Pozos R S
Published: January 2020

Concanavalin A and wheat germ agglutinin are capable of preventing a productive peripheral infection of dissociated rat sensory neurons in culture by herpes simplex virus type 1. Concanavalin A binds to the herpes simplex virion, rendering it inactive, whereas wheat germ agglutinin binds to the peripheral neuritic extensions of the sensory neurons, rendering them incapable of initiating a productive viral infection. This latter effect (i) seems to be specific for wheat germ agglutinin since other lectins have no effect, (ii) is not the result of cellular cytotoxicity, (iii) is dependent on an N-acetylneuraminic acid moiety, and (iv) may be due either to viral receptor site masking or to binding of wheat germ agglutinin to the neuritic receptor molecule for herpes simplex virus.
Some characteristics of heavy and light bands of Rickettsia prowazekii on Renografin gradients
Authors: Hanson B A , Wisseman C L , Waddell A , Silverman D J
Published: January 2020

Suspensions of partially purified Rickettsia prowazekii yielded two bands of organisms when centrifuged to equilibrium in Renografin density gradients. Rickettsiae from the lower, heavy band were defective in their infective and metabolic activities, as compared to organisms from the light band. The greater density in Renografin of heavy-banding organisms was due to their lack of permeability barrier to it, as evidenced by the absence of plasmolysis in hypertonic Renografin. In contrast, light-banding rickettsiae were able to exclude Renografin, since they were plasmolyzed in it. The proportion of heavy-banding organisms in a rickettsial suspension was influenced by the growth phase they were in when harvested from infected yolk sacs, as well as by the conditions and media to which they subsequently were exposed. We have concluded that these defective forms arise from the degeneration of light-banding rickettsiae. This separation of two functional classes of rickettsiae in Renografin density gradients has been exploited (i) to increase the uniformity of the suspensions by removing many noninfectious particles and (ii) to determine rapidly the integrity of certain properties of the cytoplasmic membrane of organisms exposed to a variety of conditions.
Clinical microbiological, and histological manifestations of Streptobacillus moniliformis-induced arthritis in mice
Authors: Savage N L , Joiner G N , Florey D W
Published: January 2020

Intravenous inoculation of Streptobacillus moniliformis into mice resulted in an infection in which the predominant feature was progressive polyarthritis that rendered some joints immobile within 6 months. No migration of arthritis from joint to joint or remission and exacerbation were apparent. Viable organisms were apparently removed by the host from blood, liver, and spleen within 28 days post inoculation but persisted in joints for approximately 6 months in some animals. Specific antibody was detectable by complement fixation 7 days post-inoculation and persisted throughout the course of the disease. The inflammatory responses, which was initiated by the appearance of neutrophils in the joint space within 24 h of inoculation, culminated in obliteration of the joint space by fibrosis and exostosis.
Blastogenic responses of lymphocytes from mice immunized by sublethal infection with yeast cells of Histoplasma capsulatum
Authors: Tewari R P , Khardori N , McConnachie P , von Behren L A , Yamada T
Published: January 2020

Blastogenic responses of spleen cells to histoplasmin and ribosomal antigens and to the mitogens concanavalin A. phytohemagglutinin, and lipopolysaccharide were studied in normal and immunized mice (10(5) live yeast cells of Histoplasma capsulatum given by the subcutaneous route). Cells (10(6) per well) were cultured with and without antigens and mitogens in microtiter plates with RPMI 1640-5% heat-inactivated normal mouse serum for 72 h at 37 degrees C. Cells were harvested after a 16- to 18-h pulse with 1 microCi of [3H]thymidine (6.7 Ci/mol), and thymidine incorporation was measured by scintillation counting. The initial blastogenic response to concanavalin A (54 X 10(3) cpm) was suppressed (P less than 0.001) from 4 to 14 days post-immunization and returned to control levels on day 21. The response to phytohemagglutinin was suppressed up to 21 days. Lipopolysaccharide responses, however, were affected to a lesser degree. Blastogenic responses to histoplasmin and H. capsulatum ribosomes were similar on day 0 in normal and immune lymphocytes, but by day 4 cells from immunized mice were more responsive (P less than 0.01). The maximum response to H. capsulatum antigens was detected on day 42 and was 9- to 16-fold higher than in controls. These results demonstrate in vitro responses of primed lymphocytes on exposure to H. capsulatum antigens and suppressed responses to mitogens during early stages of the immune response.
Secretory and systemic immunological response in children infected with live attenuated influenza A virus vaccines
Authors: Murphy B R , Nelson D L , Wright P F , Tierney E L , Phelan M A , Chanock R M
Published: January 2020

An enzyme-linked immunosorbent assay was used to measure isotype-specific antibody to purified hemagglutinin (HA) of influenza A virus, using serum and nasal-wash specimens from young children undergoing primary infection with live cold-adapted influenza A/Alaska/77 (H3N2) or A/Hong Kong/77 (H1N1) candidate vaccine virus. The serum antibody response followed the pattern expected for a primary viral infection. Each of 17 vaccinated children had a serum immunoglobulin G (IgG) HA antibody response, 16 had an IgM antibody response, and 13 had an IgA antibody response. Nasal-wash HA antibody was detected in the IgA, IgM, and IgG isotypes. Of the 17 vaccinated children, 14 had an IgA response, 13 had an IgM response, and 9 had an IgG response. Most of the IgA and IgM HA antibody was actively secreted locally, whereas only some of the IgG HA antibody could be shown to be actively secreted into the respiratory tract. There was a good correlation between the level of nasal-wash antibodies measured by the HA-specific IgA enzyme-linked immunosorbent assay and by a plaque neutralization assay. These data indicate that intranasal vaccination of susceptible children with live, attenuated, cold-adapted influenza A viruses efficiently stimulates both systemic and local antibody responses.
Effect of cyclosporin A on murine natural killer cells
Authors: Gui X , Ho M , Camp P E
Published: January 2020

In mice, cyclosporin A decreased the natural killer cell-enhancing effect of two interferon inducers, infective murine cytomegalovirus and nonreplicating Newcastle disease virus. It also inhibited murine cytomegalovirus replication at doses greater than 20 mg/kg, but it did not significantly inhibit interferon induction by Newcastle disease virus. In cell culture, cyclosporin A had no direct effect on the natural killer activity of spleen mononuclear cells derived from normal or murine cytomegalovirus-infected animals. However, at 50 micrograms/ml it significantly reduced the ability of interferon to enhance the natural killer activity of normal spleen cell suspensions. The inhibitory effect of cyclosporin A on natural killer cell activity in infected mice may be partly explained by its ability to block the action of interferon.
Adhesion in vitro and in vivo associated with an adhesive antigen (F41) produced by a K99 mutant of the reference strain Escherichia coli B41
Authors: Morris J A , Thorns C , Scott A C , Sojka W J , Wells G A
Published: January 2020

A K99-negative mutant of the K99 reference strain Escherichia coli B41 (O101:K99) was isolated (strain B41M). It did not react with OK antiserum to a K12 K99+ recombinant or with OK antiserum to K99-positive organisms from the O8, O20, or O64 serogroups, but it did react with OK antiserum to K99-positive organisms from the O101 and O9 serogroups. The mutant hemagglutinated sheep erythrocytes and attached in vitro to calf enterocytes when cultured at 37 degrees C but not when grown at 18 degrees C. Attachment was mannose resistant but susceptible to heating and formaldehyde. These properties were associated with the presence of fimbriae. The isolated hemagglutinin migrated to the anode in immunoelectrophoresis experiments, competitively inhibited attachment of strain B41M to calf enterocytes, and could be demonstrated adhering to these cells in vitro by indirect immunofluorescent staining. The anionic hemagglutinin is referred to provisionally as F41. Germfree piglets infected with strain B41M developed diarrhea within 16 h. Scanning electron microscopy showed groups of bacteria adherent to the microvilli of villous enterocytes. Indirect immunofluorescent staining demonstrated the presence of F41 antigen in vivo.